2017 Annual Report to the Carver Trust

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ROY J. AND LUCILLE A. CARVER COLLEGE OF MEDICINE

2017 ANNUAL REPORT TO THE CARVER TRUST

2017


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TABLE OF CONTENTS 02

To the Carver Trust

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2016 Highlights

21

Endowed Core Research Facilities

35

Endowed Chairs and Professorships

81

Carver Research Programs of Excellence

105

Carver Medical Research Initiative Grants

133

Carver Young Investigator Awards

153

Financial Summary

161

College Leadership

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Dear Members of the Carver Trust Board: The year 2016 will certainly be remembered as one of the most significant in the history of the research enterprise at the University of Iowa Roy J. and Lucille A. Carver

TO THE CARVER TRUST —

College of Medicine. With the $45 million grant from the Roy J. Carver Charitable Trust and creation of the Iowa Neuroscience Institute, we are poised to make revolutionary discoveries and ultimately develop new treatments and cures for some of the most devastating diseases of the brain. The Carver Trust grant played an instrumental role attracting the Institute’s first director, Dr. Ted Abel, a highly distinguished neuroscientist who joined us in January 2017. Dr. Abel’s vision is to build a campus-wide neuroscience community whose excellence in scientific and academic distinction will rival the best anywhere in the world. The grant also stands out as the largest single contribution in For Iowa. Forever More: The Campaign for the University of Iowa. In total, the campaign raised more than $1.975 billion to help the UI remain at the forefront of education, research and health care. The Carver Trust is a true partner for the institution and its total commitment of $71 million will help ensure our success. To further escalate the academic and research success of the Carver College of Medicine, UI Health Care has committed a total of $100 million ($20 million a year for five years) to recruit new faculty, support needed resources toward successful NIH grants, and fund pilot grants. Since launching the initiative in late 2015, we have already recruited seven new research faculty members, all of whom come to the Carver College of Medicine with significant NIHfunded grants. Two of the researchers, Drs. Ted Abel and Michael Tomasson, are already on campus and the others will arrive and begin work in the next three to six months. This year overall external funding has increased for the Carver College of Medicine, with more than $229.4 million received in Fiscal Year 2016—a 2.46 percent increase over FY 2015. Contributing significantly to that increase was a 13.5 percent increase ($13.6 million) in grants from the National Institutes of Health, for a total of $117.1 million in NIH funding for FY 2016. These sources of funding have allowed us to make significant progress in our five major translational pillars of research:

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neuroscience; diabetes, metabolism, and obesity; cancer;

Through our planning efforts and the creative thinking

heart and lung; and immunity, inflammation, and infection.

of some of our most talented people, we believe we

Summaries of some of the significant research findings begin

have developed a robust yet flexible plan that will

on page 6.

allow us to continue to be successful in carrying out our tripartite mission and achieving our goals.

Research Vision and Mission

This year, we will be exploring opportunities to expand our informatics capabilities and enhance the

In 2017 University of Iowa Health Care will begin

success of our immunology, infectious disease and

implementing a new, three-year strategic plan, focusing on

inflammation researchers. We are also creating a new

initiatives that are most important to our future success.

metabolomics research core and Dr. Abel is developing a neurophenotyping core. We fully expect these two core

Our strategic-planning efforts ensured that we explored

facilities to enhance the success of our top scientists.

the many possible futures that may unfold and that we considered how our path forward might change—or not

It has been a privilege to work with the Roy J. Carver

change—in response. We developed strategic storylines

Charitable Trust, the Trust staff and Board of Trustees,

that explored how the landscape of academic medicine,

and the Carver Family over these many years in support of

including our research mission, will evolve over the next

biomedical research at the Carver College of Medicine. The

decade, and we considered how global economic growth,

impact of that support is reflected in the game-changing

consumerism, personalized medicine, and the emergence of

research being conducted here every day, allowing us to deliver

big data and other technologies will permeate all aspects of

on our mission of changing medicine and changing lives.

what we do. The whirlwind pace of change in science and medicine is the common thread that requires us to build

Sincerely,

capacity to manage that change and enable our organization to be more responsive and flexible than ever before. It is clear that reinforcing and advancing our tripartite mission is our key point of differentiation. We have an overarching strategic imperative to invest, align, and integrate clinical, research, and education activities; establish a clear plan of economic interdependency between elements of the tripartite

Jean Robillard, MD

mission; prioritize research linked to clinical strategies; and

Vice President for Medical Affiars

prepare our workforce for an evolving health system.

& Dean, Carver College of Medicine

For the research enterprise, we identified the following priorities and areas for action in the three-year plan: • Develop organizational capabilities for the shift to new models of research • Align and integrate research with the clinical mission and patient outcomes • Invest in data expertise and infrastructure

Patricia Winokur, MD Executive Dean Associate Dean for Clinical and Translational Science Professor, Department of Internal Medicine

as high priority, core support • Expand sources of funding

TO T H E C A R V E R T R U S T

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2016 HIGHLIGHTS

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BREAKTHROUGHS IN MEDICINE

In the following pages, you will find a summary of important activities over the past year for UI Health Care and the University of Iowa Carver College

using CRISPR/Cas9, a new gene-editing technology. The technique is so precise it corrected a single DNA change that had damaged the RPGR gene. And because the corrected tissue had been derived from the patient’s own cells, it could potentially be transplanted without the need for anti-rejection drugs. The research was done in conjunction with scientists

of Medicine. These activities include

at Columbia University Medical Center, and published in the

breakthroughs in medicine, research award

journal Scientific Reports.

highlights, major philanthropic research gifts, and professional honors and awards to our faculty. We also highlight administrative changes in the college and three faculty members in memoriam.

MAKING PROGRESS TOWARD GENE THERAPY FOR CYSTIC FIBROSIS Two studies from the University of Iowa suggest that gene therapy may be a viable approach for treating or preventing lung disease caused by cystic fibrosis. Working with Cystic Fibrosis (CF) pigs, two teams tested two different gene

EDITED STEM CELLS OFFER HOPE OF PRECISION

therapy strategies to get the cystic fibrosis transmembrane

THERAPY FOR BLINDNESS

conductance regulator (CFTR) into the airway cells of CF pigs. One group, led by Paul McCray Jr., MD, Professor

Alexander Bassuk, MD, PhD, Professor of Pediatrics and Neurology, and Vinit Mahajan, MD, PhD, Clinical Assistant Professor of Ophthalmology and Visual Sciences, led a team of researchers who have corrected a blindnesscausing gene mutation in stem cells derived from an affected patient. The result offers hope that eye diseases might one day be treated with patients’ own edited tissue. With the aim of repairing deteriorating retinas in patients with the inherited blinding disease X-linked retinitis pigmentosa (XLRP), Drs. Bassuk and Mahajan generated stem cells from a patient’s skin cells and then repaired the damaged gene, RPGR,

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of Pediatrics, and Patrick Sinn, PhD, Research Associate Professor of Pediatrics and Director of the UI Viral Vector Core, focused on a lentivirus. The other research team, led by Joseph Zabner, MD, Professor of Internal Medicine, and David Schaffer at the University of California, Berkeley, focused on the adeno-associated virus AAV2. The researchers showed that both gene therapies restored chloride currents in pig airway cells, indicating that both vectors delivered working CFTR to the correct location in the airways. Both approaches also increased the pH and the bacterialkilling ability of the pigs’ airway secretions. Both studies were published Sept. 8, 2016, in the journal JCI Insight.

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STUDY FINDS PROTEIN THAT CAUSES PROBLEMS FOR CF LUNGS A research team, led by Michael Welsh MD, Professor of Internal Medicine and Director of the Pappajohn Biomedical Institute, discovered an answer to why mice with cystic fibrosis (CF) gene mutations don’t develop the life-threatening lung disease that affects most people with CF. In doing so, the team also identified a proton pump that could be a target for new CF therapies. The results were published Jan. 29, 2016, in the journal Science. Viral Shah, first author on the study and student in the Medical Scientist Training Program, and his colleagues showed that in people, pigs, and mice with CF, the cystic fibrosis transmembrane conductance regulator channel is lost, stopping the flow of bicarbonate into the airways. When that happens in people and pigs, their airway liquid becomes more acidic, reducing their ability to fight infection. But in mice, the airway liquid does not become more acidic, and they are not prone to infection. That fact led the scientists to ask what secretes acid into the airways of people and pigs that is missing in the mice. They discovered that a proton pump called ATP12A is responsible.

MORE RAPID STAPH DETECTION A research team lead by James McNamara, PhD, Associate Professor of Internal Medicine, has identified a procedure that may reduce the wait for diagnosis of a staph infection— from a few days to just three hours—as well as the amount of time patients may have to wait to determine the medicine’s efficacy. One of the biggest reasons for delayed detection of staphylococcus aureus bacteremia is the exceedingly low levels of the bacteria in the blood of infected patients. Currently, blood sample testing is used to not only determine whether bacteria are in the blood, but to also determine the type of bacteria present. In their study, McNamara and his team show that by measuring a specific enzyme secreted by staph, micrococcal nuclease (MN), in a blood sample, doctors can detect and diagnose a staph infection within three hours. Based on its fast turnaround time, this enzyme test may also enable doctors to monitor whether a prescribed antibiotic is doing what it’s supposed to be doing— and change medications if it’s not, all in the same day.

2016 HIGHLIGHTS

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LINK BETWEEN CANCER AND AUTISM University of Iowa researchers have shown that patients diagnosed with an autism spectrum disorder have more cancerpromoting oncogenes but lower rates of cancer. Benjamin Darbro, MD, PhD, Assistant Professor of Medical Genetics in the Stead Family vDepartment of Pediatrics, and his team analyzed large, publicly available genomic databases of patients with autism. They found that autistic patients have significantly higher rates of DNA variation in their cancerpromoting oncogenes than those without autism. They followed up this result with an analysis of the UI Hospitals and Clinics’ electronic medical record and discovered that patients diagnosed with autism were also less likely to be diagnosed with cancer. The study was published In April 2016 in the journal PLOS One.

LIVER-GENERATED HORMONE REGULATES ‘SWEET TOOTH’ While sugar cravings are common, the physiological mechanisms that trigger our “sweet tooth” are not well defined. Matthew Potthoff, PhD, Assistant Professor of Pharmacology, led a study in mice showing that a hormone produced by the liver, fibroblast growth factor 21 (FGF21), suppresses the consumption of simple sugars. The researchers report that FGF21 is produced in the liver in response to high carbohydrate levels. FGF21 then enters the bloodstream, where it sends a signal to the brain to suppress the preference for sweets. The research could improve diets and help patients who are diabetic or obese. Dr. Potthoff was co-senior author on the paper, published online in the journal Cell Metabolism with Matthew Gillum, Professor at the University of Copenhagen (Denmark)

STUDY EXPLAINS OPPOSING EFFECTS OF HORMONE ON WEIGHT GAIN New research by University of Iowa scientists, led by Justin Grobe, PhD, Assistant Professor of Phramacology, helps explain how a hormone system often targeted to treat cardiovascular disease can also lower metabolism and promote obesity. The renin-angiotensin system (RAS) controls blood pressure and is important for cardiovascular health. Many of

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the drugs used to treat hypertension (high blood pressure) and

TYPE OF VITAMIN B3 SAFELY BOOSTS IMPORT-

heart failure block or inhibit the RAS. Increasing evidence

ANT CELL METABOLITE

suggests the RAS also plays a role in controlling energy balance and metabolic rate and therefore may be important in obesity. But, depending on where in the body this hormone system is operating, it can have opposing effects on weight gain. When the RAS is elevated in the brain, it increases energy expenditure by increasing resting metabolism, resulting in weight loss. However, increased activity of the RAS circulating in the body (the peripheral RAS)—which occurs during obesity in humans and experimental animals—has the opposite effect, decreasing resting metabolism and increasing weight gain. The work was published in the journal Cell Reports.

Led by Charles Brenner, PhD, Professor and Roy J. Carver Chair of Biochemistry, the first controlled clinical trial of a newly discovered form of vitamin B3, nicotinamide riboside (NR), has shown that the compound is safe for humans. The study also shows NR increases levels of a cell metabolite that is critical for cellular energy production and protection against stress and DNA damage. Studies in mice have shown that boosting the levels of this cell metabolite—nicotinamide adenine dinucleotide, known as NAD+—can produce multiple health benefits, including resistance to weight gain, improved control of blood sugar and cholesterol, reduced nerve damage,

HEART-DAMAGING PATHWAY TRIGGERED BY

and longer lifespan. Levels of NAD+ diminish with age, and

INSULIN

other studies have suggested that loss of this metabolite may

Cardiovascular disease is the leading cause of death in

play a role in age-related health decline. The research was reported Oct. 10, 2016, in the journal Nature Communications.

people with diabetes, and risk for heart failure—where the heart can’t pump enough blood—is two to three times higher in men and up to five times higher in women with

TIMING MAY BE KEY TO UNDERSTANDING IN

diabetes compared to people without diabetes. But new

PARKINSON’S DISEASE

research from the University of Iowa and the University of California at Davis finds that two different drugs—a beta blocker and an antidepressant—might both have potential for preventing or treating heart failure by blocking an insulin signaling pathway in the heart muscle. Previous research points to high levels of insulin, a hallmark of Type 2 diabetes, as a key factor in heart failure associated with diabetes. The new study, led by E. Dale Abel, MD, PhD, Chair and DEO of Internal Medicine, and Kevin Xiang at UC Davis, shows that too much insulin in the blood (hyperinsulinemia) contributes to heart failure by triggering a molecular chain reaction that damages heart muscle cells. Using mice with diabetes, the study traced the links in this chain reaction—a newly discovered insulin signaling pathway in heart muscle cells. Blocking the pathway with the beta blocker carvedilol or the antidepressant paroxetine reverses heart muscle damage and restores heart function in the mice. The findings, published Nov. 4, 2016, in the journal Circulation, suggest that these drugs might have potential for preventing or treating heart failure associated with Type 2 diabetes.

2016 HIGHLIGHTS

The ability to consciously guide movements over a timeframe of a few seconds is a simple but universal thinking skill in mammals. It also is an ability that is consistently impaired in patients with Parkinson’s disease (PD). In a new study, led by Nandakumar Narayanan, MD, PhD, Assistant Professor of Neurology, researchers showed for the first time that brain stimulation of specific neurons at a specific frequency can improve timing in mice that are missing dopamine. Working in the lab and the clinic, Narayanan and his team recorded brain activity in mice and humans as they did simple timing tasks. The scientists also use various genetic technologies to manipulate and study brain activity in the mice. These experiments helped to reveal the neurocircuitry that controls timing ability, and explain how this simple cognitive process is disrupted by a lack of dopamine. The findings imply that, at least in theory, it might be possible to use brain stimulation to improve cognitive problems caused by PD, and possibly other cognitive disorders. The study was published online in the journal Current Biology.

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STUDY CONTRADICTS LONG-HELD BELIEF Michael Abramoff, MD, PhD, Professor of Ophthalmology and Visual Sciences is senior author and Elliott Sohn, MD, Associate Professor of Ophthalmology and Visual sciences, is a first author on a study that turns the tables on the conventional theories about retinal diabetic neuropathy. Scientists have long known that patients with diabetes mellitus—both Type 1 and Type 2—are at high risk for developing diabetic retinopathy, which is the most common cause of irreversible blindness in adults. Vision loss occurs because of microvascular damage to the retina. People with diabetes are typically not aware that they are also at risk for developing retinal diabetic neuropathy, which is the loss of nerve cells in the retina. For many years, scientists believed patients developed retinopathy and, as a result of the damage to the blood vessels, later developed neuropathy. Doctors were focusing on early detection and treatment of retinopathy to prevent blindness, which they thought would then prevent the damage caused by neuropathy. However, in the online journal PNAS, Drs. Abramoff and Sohn reported that the sequence of events occurring in the retina is just the opposite. They found that the nerve damage comes first, before the vessel damage; and that even people with diabetes who never get retinopathy can develop this damage, and after many years, the damage may be severe and similar to glaucoma.

NEW MUSCLE-WASTING PROTEIN FOUND In a new study published in the Journal of Biological Chemistry, Christopher Adams, MD, Professor of Internal Medicine, and colleagues at the UI and the Buck Institute for Research on Aging continued their work investigating a protein, Gadd45a (Growth Arrest and DNA Damage-inducible 45 alpha), in mice. By identifying other proteins that interact

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with Gadd45a during muscle atrophy, the scientists found a key partner for Gadd45a in the process, a muscle enzyme called MEKK4—a protein that had not previously been linked to muscle wasting. To find the proteins that interact with Gadd45a, they developed a new biochemical method and used it to pull out this protein’s partners in crime to put together a molecular picture of how muscle atrophy happens, with the goal of finding a way to intervene and create therapies that can prevent or treat this serious condition.

GENETIC OVERLAPS FOUND IN PSYCHIATRIC DISEASES

FINDINGS IN TRAUMATIC BRAIN INJURY

James Potash, MD, MPH, Professor and Chair and DEO

Researchers from the University of Iowa, led by Andrew

of Psychiatry is the senior author of a study suggesting there

Pieper, MD, PhD, Professor of Psychiatry, believe they

may be an overlap between rare genetic variations linked to

may have identified a potential approach for preventing

bipolar disorder (BD) and those implicated in schizophrenia

the development of neurological problems associated with

and autism. The study, by researchers at the University of

traumatic brain injury (TBI). Their research in mice suggests

Iowa, Johns Hopkins University School of Medicine, and

that protecting axons—the fiber-like projections that connect

Cold Spring Harbor Laboratory, was published in JAMA

brain cells—prevents the long-term neuropsychiatric problems

Psychiatry and adds to the growing understanding that many

caused by blast-related traumatic brain injury. In a recent

psychiatric diseases share genetic roots. It is also among the

study, the UI team investigated whether early damage to

first to suggest a genetic overlap between BD and autism.

axons—an event that is strongly associated with many forms of brain injury, including blast-related TBI—is simply a consequence of the injury or whether it is a driving cause of

INFUSING STEM CELLS HELPS AVOID FLUID

the subsequent neurological and psychiatric symptoms.

BUILDUP IN EYES A team of researchers from the University of Iowa and Veterans Affairs led by Markus Kuehn, PhD, Associate Professor of Ophthalmology and Visual Sciences, recently published its work on infusing stem cells to help restore proper drainage for fluid-clogged eyes at risk for glaucoma in the Proceedings of the National Academy of Sciences. The influx of cells regenerated the tiny, delicate patch of tissue known as the trabecular meshwork, which serves as a drain for the eyes to avoid fluid buildup. When fluid accumulates in the eye, the increase in pressure can lead to glaucoma. The disease damages the optic nerve and can result in blindness. One potential advantage of the approach is that the type of stem cells used—called induced pluripotent stem cells—could be created from cells harvested from a patient’s own skin.

2016 HIGHLIGHTS

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RESEARCH AWARD HIGHLIGHTS The Combating Antimicrobial Resistance through Rapid

R01 research grant entitled “Role of PPARγ and the PPARγ

Implementation of Available Guidelines and Evidence

Target Gene RBP7 in the Endothelium,” to determine how

(CARRIAGE) study, led by program directors Eli

RBP7 (retinol binding protein 7) acts as a specific co-factor for

Perencevich, MD, MS, Professor of Internal Medicine and

PPARγ in the endothelium, the inner most lining of the blood

Epidemiology, and Dr. Michael Rubin from University of Utah

vessel, to protect the vessel from oxidative stress which can

Health Care, has received funding through the Department of

occur in response to stressors such as high fat diets and aging.

Veterans Affairs (VA) Quality Enhancement Research Initiative (QUERI). The QUERI program’s goal is to improve the

Chioma Okeoma, PhD, Assistant Professor of

health and care of Veterans by implementing effective clinical

Microbiology, received a five-year $3.1 million

practices into routine care. The CARRIAGE QUERI aims

National Institutes of Health R01 research grant

to address the growing concern of antimicrobial resistance

entitled “The effect of HIV and cocaine abuse on

through strategies targeting improved use of antibiotics

semen exosome composition and function”.

and prevention of transmission of antimicrobial resistant bacteria within VA patient care settings.

François Abboud, MD, Professor of Internal Medicine, has received a five-year renewal of the T-32 grant that

Eric Hoffman, PhD, Professor of Radiology, received a five-

has funded the Cardiovascular Institutional Research

year $4.3 million National Institutes of Health R01 research

Fellowship, a postdoctoral program providing mentorship,

grant entitled “Functional CT Assessment of Pulmonary

guidance, and support to physicians and basic scientists in

Arterial Dysfunction in Smoking Associated Emphysema”.

cardiovascular medicine. Graduates of this fellowship are distinguished investigators and leaders at the University

Stanley Perlman, MD, PhD, Professor of Microbiology,

of Iowa and around the world. Their discoveries have led

received a five-year $2.7 million National Institutes

to a greater understanding and more skillful management

of Health R01 research grant entitled “Role of

of heart disease and stroke. This program has been

eicosanoids in pathogenic human CoV infections”.

continuously funded by the National Heart, Lung, and Blood Institute since 1975 and will continue through 2021.

Val Sheffield, MD, PhD, Professor of Pediatrics, received a five-year $2.6 million National Institutes of Health P30 award

Paloma Giangrande, PhD, Professor of Hematology,

to provide infrastructure for the inherited eye disease research

Oncology, and Blood and Marrow Transplantation, has

performed in their laboratories and the laboratories of their

been awarded a two-year, $1.1 million grant from the

collaborators in the Wynn Institute for Vision Research.

Department of Defense to study potential treatments for certain instances of blunt chest traumas. In some cases,

Curt Sigmund, PhD, Professor and Chair of Pharmacology,

two resulting syndromes—multiorgan dysfunction and

received a four-year $3.0 million National Institutes of Health

acute respiratory distress—produce toxic extracellular

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histones, which contribute to patients’ high morbidity

Protocol and the previously developed Swank diet to

and mortality rates. Previously developed RNA aptamers

treat MSrelated fatigue, a disabling symptom that can

have been shown to bind to and neutralize their effects.

significantly interfere with a person’s ability to function at

Giangrande and her team will evaluate the efficacy of

home and work. The grant is one of the largest financial

these bio-drugs in preventing histone-mediated cell injury

commitments made by the NMSS to research this year.

in cultures and evaluate their efficacy in animal models. George Weiner, MD, Professor and Director of the Nitin Karandikar, MD, PhD, Chair and DEO of

Holden Comprehensive Cancer Center, and Aliasger

Pathology, received grant funding of $3.6 million from the

Salem, PhD, Bighley Professor of Pharmaceutical

National Institute of Health/National Institute of Allergy

Sciences, received a $600,000 grant from the Leukemia

and Infectious Diseases (NIH/NIAID) for a project titled

and Lymphoma Society to further their research to

“Immune Dysregulation During Multiple Sclerosis Relapse”.

develop nanoparticles that can be injected directly into the

Dr. Karandikar’s laboratory is focused on understanding

lymphoma cells to create a type of immunotherapy. The

the immune processes that underlie the pathogenesis and

research has two specific aims: develop the nanoparticles

regulation of multiple sclerosis, an immune-mediated

and test nanoparticles’ ability to trigger a response in

demyelinating disease of the central nervous system.

lymphoma when used alone and with other therapies.

The University of Iowa Research Foundation has finalized

Phil Polgreen, MD, Associate Professor of Infectious

a license and sponsored research agreement with Pfizer Inc.

Diseases, has been awarded an exploratory/developmental

to support the development of potential gene therapies for

research (R21) grant from the National Institute of Diabetes,

cystic fibrosis (CF) by the laboratories of John Engelhardt,

Digestive, and Kidney Diseases. These NIH funds will fuel

PhD, Professor and Chair of Anatomy and Cell Biology, and

a novel approach to encouraging individuals with Type 2

Ziying Yan, PhD, Research Associate Professor of Anatomy

diabetes (T2D) and those at risk of acquiring the disease

and Cell Biology. Pfizer will collaborate with the doctors to

to decrease their sedentary time, a cornerstone of T2D

develop a potential unique gene therapy for cystic fibrosis.

prevention and management. Polgreen has been working with Alberto Segre, PhD, Professor of Computer Science, in

The American Heart Association has designated the

developing a phone-based walking game that allows people

University of Iowa as a Strategically Focused Heart Failure

to take a virtual walk through interesting global locations

Network and provided more than $3.8 million for its work.

at the same time as their real-world steps are counted by

Approximately $1 million of that will fund the lab of E. Dale

a wrist-worn accelerometer. Users of this application,

Abel, MD, PhD, Chair and DEO of Internal Medicine, as

called MapTrek, can monitor their own progress and be

members perform the basic science portion in partnership

encouraged to make daily use of it, thus helping reduce

with researchers at the University of Utah. They will test the

their risk of T2D and better manage their disease.

hypothesis that improving the efficiency of cardiac glucose metabolism may predict cardiac recovery in heart failure.

The Leadership Education in Neurodevelopmental

This is the second AHA-funded strategically focused network

and other Related Disabilities (LEND) program

to come to the University of Iowa. Curt Sigmund, PhD,

at the UI Stead Family Children’s Hospital Center for

Chair and DEO of the Department of Pharmacology,

Disabilities and Development has received a five-year, $3.1

leads one of four networks studying hypertension.

million award from the Health Resources and Services Administration. LEND programs prepare trainees from

Terry Wahls, MD, Professor of Internal Medicine,

diverse professional disciplines to assume leadership

knows firsthand what the right diet can do for a person’s

roles and ensure high levels of interdisciplinary clinical

health and well-being; her own diet, the Wahls Protocol,

competence to improve the health of infants, children,

helped her combat the fatigue and physical symptoms of

adolescents, and young adults with neurodevelopmental

multiple sclerosis (MS) nearly 10 years ago. Now, Wahls

and other related disabilities including autism.

has been awarded a $1 million grant from the National Multiple Sclerosis Society (NMSS) to compare the Wahls

2016 HIGHLIGHTS

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PROFESSIONAL AWARDS AND HONORS Christopher Ahern, PhD, Associate Professor of

study section for the National Institutes of Health

Molecular Physiology and Biophysics, was selected as a

(NIH) Center for Scientific Review on July 1.

member of the Biophysics of Neural Systems Study Section, Center for Scientific Review, from the National Institutes

An abstract by Mark Madsen, PhD, Professor of

of Health (NIH) for a six-year term beginning July 1.

Radiology, was chosen to receive the 2016 Henry Wagner, Jr., MD, Best Abstract Award at the Society of Nuclear

Wallace (Lee) L. M. Alward, MD, Frederick C.

Medicine and Molecular Imaging’s annual meeting

Blodi Chair in Ophthalmology, was selected as the inaugural recipient of the 2016 American Glaucoma

Vinit Mahajan, MD, PhD, Clinical Assistant Professor

Society Outstanding Educator Award in recognition of

of Ophthalmology and Visual Sciences, received the 2016

his extraordinary commitment to advancing glaucoma

Alumni Achievement Award from Fight for Sight.

education to residents, fellows, and colleagues. Chioma Okeoma, PhD, Assistant Professor of Michael Anderson, PhD, Associate Professor of

Microbiology, was recognized with the Research Innovation

Ophthalmology and Visual Sciences, will serve as a

and Leadership Award from the Technology Association

member of the Biology and Visual System Study Section,

of Iowa. Dr. Okeoma studies cellular tactics used by our

Center for Scientific Review, from the National Institutes

bodies (and those of other mammals) to fight off viruses.

of Health for a four-year term beginning July 1, 2017.

She was one of eight women leaders in the fields of science, technology, engineering, and math (STEM) to be

Charles Brenner, PhD, Professor and Chair and

honored as winners of the ninth annual Des Moines Area

DEO of Biochemistry, received the 2016 American

Community College Iowa Women of Innovation Awards.

Society for Biochemistry and Molecular Biology Award for Exemplary Contributions to Education.

UI Carver College of Medicine student, Thomas Pak, was chosen by the Howard Hughes Medical Institute

Kevin Campbell, PhD, Professor and Chair and DEO

(HHMI) as a medical fellow. The HHMI awarded

of Molecular Physiology and Biophysics, was elected

fellowships to 66 medical and veterinary students that

as a Lifetime Achievement Fellow by The American

will allow them to spend a year in a laboratory working

Society for Cell Biology (ASCB). Fellows are elected

on a research project that each student proposed.

on the basis of a career of exceptional contributions to science and for their service to the ASCB.

Eli Perencevich, MD, MS, Associate Chair for Clinical and Health Services Research and Professor of

Anil Chauhan, PhD, Associate Professor of Internal

General Internal Medicine and Epidemiology, has been

Medicine, joined the hemostasis and thrombosis

elected to the American Society for Clinical Investigation

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(ASCI). For more than 100 years, the ASCI has counted the most respected leaders in academic medicine in its 3,000-member body. These physician scientists from every medical specialty have achieved membership by virtue of their excellence in biomedical research. Andrew Pieper, MD, PhD, Professor of Psychiatry, was selected for membership in the American Society for Clinical Investigation (ASCI). Dr. Pieper’s election brings the number of UI faculty among the ASCI membership to 34. Kamal Rahmouni, PhD, Professor of Pharmacology and Internal Medicine, received the 2016 Paul Korner Award from the International Society of Hypertension. The award, recognizes scientists who have made outstanding contributions to hypertension research in the field of neuroscience. Richard Shields, PhD, Chair and DEO of Physical Therapy and Rehabilitation Science, is the recipient of the 2017 Mary McMillan Lecture Award from the American Physical Therapy Association. The award is the highest honor provided by the organization to a physical therapist. Edwin M. Stone, MD, PhD, Professor of Ophthalmology and Visual Sciences and Director of the Steven A. Wynn Institute for Vision Research, was named to The Power List 2016 by The Ophthalmologist online publication. The list recognizes the Top 100 most influential people in the world of ophthalmology. George Weiner, MD, Professor of Internal Medicine and Director of the UI Holden Comprehensive Center, and Paul McCray, MD, Professor of Pediatrics and Microbiology, were chosen by the UI Faculty Senate to receive a 2016 Regents Award for Faculty Excellence. Michael Welsh, MD, Professor of Internal Medicine, has been named the recipient of the 2017 Steven C. Beering Award. The Indiana University School of Medicine Steven C. Beering Award honors an internationally recognized individual for outstanding research contributions to the advancement of biomedical or clinical science. Seven of the past Beering Award recipients have become Nobel laureates, either before or after receipt of this award. John Wemmie, MD, PhD, served as President of the Psychiatry Research Society.

2016 HIGHLIGHTS

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NEWLY APPOINTED ENDOWED CHAIRS & PROFESSORSHIPS

Ted Abel, PhD, was named the Roy J. Carver Chair in Neuroscience. Colin Derdeyn, MD, was named the Kenneth L. and Gloria D. Krabbenhoft Chair in Radiology.

ADMINISTRATIVE CHANGES

Jeremy Greenlee, MD, was named the Arnold H. Menezes Chair in Neurosurgery. Scott Larson, MD, was invested as the William E. Scott

James Choi, MD, Associate Professor of Anesthesia, and Amal Shibli-Rahhal, MD, Clinical Associate

Educational Professor in Pediatric Ophthalmology.

Professor of Endocrinology, and were appointed

Mohammed Milhem, MBBS, was named the

Office of Student Affairs and Curriculum.

Holden Family Chair in Experimental Therapeutics. Robert Piper, PhD, was named the Roy J. Carver Professor in Molecular Physiology and Biophysics. Margo Schilling, MD, was named the John McMaster

assistant deans in the UI Carver College of Medicine

Jean E. Robillard, MD, was named Dean of the UI Roy J. and Lucille A. Carver College of Medicine (and maintains his role as Vice President for Medical Affairs), effective Feb. 15, 2016.

Ristine, MD, Geriatric Education Professor.

Marta Van Beek, MD, MPH, FAAD, Clinical

Michael Wagoner, MD, PhD, was invested as

UI Hospitals and Clinics Chief of Staff, and

Beulah and Florence Usher Chair in Cornea/ External Disease and Refractive Surgery. John Wemmie, MD, PhD, was named the Roy J.

Associate Professor of Dermatology, was elected began serving in that role Jan. 11, 2016. Patricia Winokur, MD, was named Executive Dean of the UI Carver College of Medicine, effective Jan. 1, 2016. As

Carver Chair in Psychiatry and Neuroscience.

Executive Dean, Dr. Winokur works closely with Dean Jean E.

Brian Wolf, MD, MS, Professor of Orthopedics

clinical, and academic affairs. In addition, she serves as Senior

and Rehabilitation, has been appointed the John

Robillard in overseeing our missions of research, education, Associate Dean for Clinical and Translational Science.

and Kim Callaghan Chair in Sports Medicine.

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MAJOR PHILANTHROPIC RESEARCH GIFTS*

In February 2017 the University of Iowa and the University of Iowa Foundation announced the successful conclusion of For Iowa. Forever More: The Campaign for the University of Iowa. More than 272,000 UI alumni and friends—enough to fill Kinnick Stadium nearly four times—contributed more than $1.975 billion to help the UI remain at the forefront of education, research and health care. For UI Health Care, more than 146,600 donors contributed over $834 million. Their giving supports students, faculty, facilities, programs, and research in the UI Roy J. and Lucille A. Carver College of Medicine and UI Hospitals and Clinics. The Stephen A. Wynn Institute for Vision Research received significant support for Usher IIIa, Glaucoma, and Macular Degeneration research through commitments of $2 million from the Prince Family Foundation, $1.5 million from Francis and Muriel Jeffries, and $1 million from the Elmer & Sylvia Sramek Charitable Foundation, respectively. Francis and Muriel Jeffries provided additional support through a $1.5 million allocation to cancer research in the Holden Comprehensive Cancer Center. *In addition to those provided by the Roy J. Carver Charitable Trust

2016 HIGHLIGHTS

17


IN MEMORIUM

RICHARD KERBER, MD

REX MONTGOMERY, MD Rex Montgomery, MD, Professor Emeritus of Biochemistry, died July 31, 2016, as he neared his 93 birthday. Dr. rd

Montgomery was the longest serving member of the department of Biochemistry, a great leader at the University of Iowa, an outstanding carbohydrate biochemist, a devoted father and grandfather, and a wonderful human being. Dr. Montgomery joined the Iowa faculty as an Assistant Professor in 1955, and became a full Professor in 1963. He also served as Associate Dean for academic affairs in the Carver College of Medicine from 1974 to 1995, during which time he also held the positions of Associate Dean of research in the CCOM and Interim Vice President of Research for the University. His research and scholarly efforts had a major global impact. Two of his textbooks, described as influential and strikingly important, transformed biochemistry education. He was also admired and appreciated for his remarkable impact as a teacher and mentor. In 1974, Dr. Montgomery established a new physician assistant program at the University of Iowa, and served as its director until 1976.

18

Richard Kerber, MD, Professor of Cardiovascular Medicine, died following a short illness at UI Hospitals and Clinics on Nov. 8, 2016. He was 77. For more than 45 years Dr. Kerber dedicated himself to the University of Iowa, demonstrating a loyalty to and belief in the institution’s mission rarely found in others. He was a distinguished academic physician, and for nearly 30 years served as Associate Director of the Division of Cardiovascular Medicine, only leaving the position to serve as Interim Director of the division from 2009-2012. As a researcher with a focus on cardiac resuscitation, he had few peers, authoring more than 250 articles, two books, more than 330 abstracts, and dozens of chapters of books. His stature in the field was recognized among other honors by his election to the American Association of Physicians in 1988. His influence on cardiovascular research and clinical innovation will be felt for years to come. Dr. Kerber was similarly dedicated to the educational mission, serving as director of the Cardiology Fellowship Program for 17 years, and regularly earning teaching awards, including the Ernest O. Theilen Clinical Teaching and Service Award in 2013.

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PAUL SEEBOHM, MD Paul Seebohm, MD, Professor Emeritus, Internal Medicine, and former Executive Associate Dean, died Jan. 8, 2017, five days short of his 100th birthday. Dr. Seebohm joined the UI Department of Internal Medicine in 1949 as the first director of a new subspecialty—the Division of Allergy and Immunology. The entirety of Dr. Seebohm’s academic career was spent at Iowa where he taught generations of innovative, successful allergists and immunologists. He directed the allergy division from 1949 to 1970, when he became Associate Dean. In 1974 he was named Executive Associate Dean for the UI College of Medicine. As Executive Dean, he was instrumental in the creation of a Family Practice Training Program, which had a major impact on the primary care of patients throughout Iowa. His contributions to academic medicine and the subspecialty of allergy and immunology were many and played out on the national scene as he served in leadership positions of many professional organizations, including the American Academy of Allergy and Immunology, which presented him with its highest honor in 1974 as Distinguished Service Awardee. In the American Medical Association (AMA), he served as secretary of the AMA Section Council on Allergy and was its chairman. He was instrumental in establishing the allergy/immunology certification board and was the first to win board certification. At the National Institutes of Health, he served on the Allergy and Immunology Research Committee and became its chair as well. He also served and eventually chaired the Food and Drug Administration’s panel on Review of Allergenic Extracts. Dr. Seebohm was president of the Iowa Medical Society, president of the Iowa State Board of Health and served on the Board of Directors of the Health Policy Corporation of Iowa and on several Governor’s Advisory boards. Though he officially retired in 1986, Dr. Seebohm remained active in the UI Department of Internal Medicine until shortly before his death.

2016 HIGHLIGHTS

19



ENDOWED CORE FACILITIES

21


The mission of the Iowa Institute of Human Genetics (IIHG) Genomics Division (formerly the DNA Facility) is to support and provide cutting-edge resources and services used in the analysis of DNA and RNA to University

IOWA INSTITUTE OF

of Iowa research and clinical investigators.

HUMAN GENETICS

Funds from the Roy J. Carver Charitable Trust were

GENOMICS DIVISION

used in 2016 to build infrastructure and develop services around the cutting-edge massively parallel or next generation sequencing (NGS) technologies. Specifically, the contributions of the Carver Trust were used to: 1. support the salaries of two technicians, Einat

KEVIN L. KNUDTSON, PHD DIRECTOR

Snir and Mari Eyestone, who prepared and ran samples on the next generation sequencers; 2. support the use of a sample handling robot to set up sequencing reactions for two new clinical tests (KidneySeqTM and drug metabolism); 3. acquire computer equipment used to analyze and store data generated by the NGS instrumentation; and 4. support the service contracts and upgrades for the NGS support instrumentation. An ongoing challenge with NGS platforms is data analysis and storage. Thomas Bair, PhD, leads a team of IIHG bioinformaticians in support of analysis needs of Genomics Division users. While the Carver Trust did not specifically support Dr. Bair this year, his team has been essential in supporting the analysis and data management, using the tools that were acquired with Carver Trust monies. Scientific Advances with the Human Genome The first human genome completed under the Human Genome Project took over ten years to finish at cost of nearly three billion dollars (~ $1 per base). Because of the advances in massively parallel or next generation sequencing (NGS) technologies, a human genome can now be sequenced in under a week at a cost of less than five cents per million bases. These unprecedented low sequencing costs have, in part, enabled investigators to now perform whole genome sequencing-based studies and support clinical precision

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and translational medicine initiatives. Over the last few

novel X-linked deletion that may be associated with autism.

years, the division has emphasized developing workflows

As highlighted by an asterisk in Figure 1, the sequence

and providing services to support investigators that are

coverage of a 10,700 base region of the mother’s X

using these rapidly evolving technologies in their studies.

chromosome was reduced by half indicating a loss of this region in one of her X chromosomes. Her identical twin

The Genomics Division has developed pipelines to

sons inherited the copy of the X chromosome containing the

work with both basic and clinical scientists especially in

deletion. The 10,700 base deletion encompasses regulatory

the area of identifying DNA sequence variations that

regions that are thought to be involved in the regulation

are associated with diseases or disorders. Currently,

of two genes, ZNF81 and ZNF182, implicated as risk

the services and resources provided include:

genes for intellectual disability and autism disorders.

1. DNA sequencing and genotyping;

The division worked closely with Edwin M. Stone, MD, PhD, Seamans-Hauser Chair in Molecular Ophthalmology,

2. custom oligonucleotides;

and Richard Smith, MD, IIHG Director and Professor of Otolaryngology, Pediatrics, Internal Medicine, Molecular

3. DNA microarray;

Physiology, and Biophysics, both in the Carver College of Medicine, to identify sequence variants associated with eye

4. molecular biology computing; 5. real-time PCR; and 6. genome sequencing. In addition, the division also provides the sequencing support for the IIHG’s three clinical NGS-based genetics tests including whole exome sequencing; a newly expanded drug metabolism test to predict tolerance to Plavix®, opioids, many drugs used to treat cancer and psychiatric disorders; and KidneySeqTM (surveys over 170 genes related to over 75 renal diseases). Notably in 2016, the division worked closely with Jacob J. Michaelson, PhD, Department of Psychiatry, to support his large-scale whole genome sequencing project of four hundred individuals to identify genome sequence variations that contribute to speech or communication disorders. Einat Snir and Mari Eyestone, Genome and Sanger Sequencing technicians, lead the efforts to optimize the sample preparation protocols and to translate the optimized workflows to the liquid handling robots to improve the processing throughput of these samples. The division can now process up to 96 genomes at a time and these improved workflows permitted the sequencing of 200 genomes, which was the year 1 aim of Dr. Michaelson’s project. Preliminary analysis of the whole genome sequence data generated for Dr. Michaelson has already uncovered a

E N D O W E D C O R E FAC I L I T I E S

disease and hearing loss, respectively. The Genomics Division has been using a sequence target capture technique that permits the specific isolation of selected genes known to be associated with a given disease from the individual’s entire genome. These “captured” genes were then sequenced and compared to a reference sequence to determine whether a sequence variation might be disease-causing. Improved efficiency of sequence target-capture sample preparation for use in clinical diagnostics was a major emphasis last year. The preparation of these samples can take a week or more to process and that greatly impacts the time it takes for investigators and clinicians to obtain their results. Mari Eyestone, leads the efforts to optimize the liquid handling robots to improve the processing throughput of these samples. In all, we processed over 450 subject samples for sequence variations associated with eye disease and over 1,000 samples for sequence variations associated with hearing loss. In summary, the Genomics Division is grateful to the Carver Trust, as its support has permitted the division to continue to provide cutting edge technologies to the University of Iowa research community. As discussed above, the new advancements in genomics and functional genomics research have similarly created new technical and analysis challenges. The Carver Trust support has been instrumental in providing the resources that permit the Genomics Division to continue to explore the best ways to meet these challenges so that researchers remain at the leading edge of scientific innovation and discovery.

23


Figure 1. Depth of sequence coverage map of the X chromosomes of identical twin sons with autism and their parents. The red dashed rectangle highlights a 10,700 base deletion. The asterisk indicates the region on the mother’s X chromosome that had half the expected depth of sequence coverage.

A novel primate-specific noncoding RNA modulates human embryo-

SELECTED PUBLICATIONS

(from at least 225 publications that involved the use of the Genomics Division in 2016)

and pluripotent stem cell fate. Nature Genetics. 48(1):44-52, 2015.

4. He B, Xing S, Chen C, Gao P, Teng L, Shan Q , Gullicksrud JA, Martin MD, Yu S, Harty JT, Badovinac VP, Tan K, Xue HH. CD8+ T Cells Utilize Highly Dynamic Enhancer Repertoires and Regulatory

1. Briggs D, Yoshida-Moriguchi T, Zheng T, Venzke D, Anderson M, Strazzulli A, Moracci M, Yu, L, Hohenester E, Campbell KP. Structural Basis of Laminin Binding to the LARGE Glycans on Dystroglycan. Nat Chem Biol. 12(10):810-814, 2016.

2. Chu Y, Lund DD, Doshi H, Keen HL, Knudtson KL, Funk ND, Shao JQ , Cheng J, Hajj GP, Zimmerman KA, Davis MK, Brooks RM, Chapleau MW, Sigmund CD, Weiss RM, Heistad DD. Fibrotic Aortic Valve Stenosis in Hypercholesterolemic/ Hypertensive Mice. Arterioscler Thromb Vasc Biol. 36(3):466-74, 2016.

3. Durruthy-Durruthy J, Sebastiano V, Wossidlo M, Cepeda D., Cui J., Grow E.J., Davila J., Mall M., Wing W.H., Wysocka J., Au, K.F., Pera, R.R.

24

Circuitry in Response to Infections. Immunity, 45(6):1341-1354, 2016.

5. Hornick AL, Li N, Oakland M, McCray PB Jr, Sinn PL. Human, Pig, and Mouse Interferon-Induced Transmembrane Proteins Partially Restrict Pseudotyped Lentiviral Vectors. Hum Gene Ther, 27(5):354-62, 2016.

6. Mishanina TV, Yu L, Karunaratne K, Mondal D, Corcoran JM, Choi MA, Kohen A. An unprecedented mechanism of nucleotide methylation in organisms containing thyX. Science, 351: 507-510, 2016.

7. Schaut, RG, Lamb, IM, Toepp, AJ, Scott, B, Jeronimo, SMB, Harty, J, Waldschmidt, T., Wilson, ME, Petersen, CA. A Novel IgDhi B Cell Population Suppresses T cell Function via IL-

2 0 1 7 A N N U A L R E P O R T TO T H E C A R V E R T R U S T


January 1, 2016 — December 31, 2016 FINANCIAL SUMMARY IOWA INSTITUTE OF HUMAN GENETICS GENOMICS DIVISION Balance Forward Revenue Disbursements to UI Total Revenue

($58,379)

Equipment Maintenance (5.6%)

$188,730

Expenses Personnel $118,470 Equipment Maintenance $7,037 Total Expenses 125,507 Ending Balance

Personnel (94.4%)

$4,844

10 and PD-L1 that Correlates with Disease Progression During Leishmania infantum Infection. J Immunol, 196(10): 4100-9, 2016.

12. Tian Y, Aursnes M, Hansen TV, Tungen JE, Galpin JG, Leisle L, Ahern CA, Xua R, Heinemann SH, and Toshinori H. Atomic determinants of BK channel activation by polyunsaturated fatty

8. Schulmeyer KH, Diaz MR, Bair TB, Sanders W, Gode CJ, Laederach

acids. Proc. Natl. Acad. Sci. 113(48):13905-13910, 2016.

A, Wolfgang MC, Yahr TL. Primary and Secondary Sequence Structure Requirements for Recognition and Discrimination of Target RNAs by Pseudomonas aeruginosa RsmA and RsmF. J Bacteriol. 25;198(18):2458-69, 2016.

13. Ulland TK, Jain N, Hornick EE, Elliott EI, Clay GM, Sadler JJ, Mills KAM, Janowski AM, Paige A, Volk D, Wang K, Legge KL, Gakhar L, Bourdi M, Ferguson PJ, Wilson ME, Cassel SL, and Sutterwala FS. Nlrp12

9. Shah VS, Meyerholz DK, Tang XX, Reznikov L, Abou Alaiwa M,

mutation causes C57BL/6J strain-specific defect in neutrophil recruitment

Ernst SE, Karp PH, Wohlford-Lenane CL, Heilmann KP, Leidinger MR, Allen PD, Zabner J, McCray PB Jr, Ostedgaard LS, Stoltz DA, Randak CO, and Welsh MJ. Airway Acidification Initiates Host Defense

14. Nature Communications 7, Article number: 13180, 2016. doi:10.1038/ncomms13180

Abnormalities in Cystic Fibrosis Mice. Science, 351(6272):503-507, 2016. 15. Zeng S, Whitmore SS, Sohn EH, Riker MJ, Wiley LA, Scheetz 10. Shao P, Liu Q , Maina PK, Cui J, Bair TB, Li T, Umesalma S,

TE, Stone EM, Tucker BA, Mullins RF. Molecular Response of

Zhang W, Qi HH. Histone demethylase PHF8 promotes epithelial

Chorioretinal Endothelial Cells to Complement Injury: Implications

to mesenchymal transition and breast tumorigenesis. Nucleic Acids

for Macular Degeneration. J Pathol 238(3):446-56, 2016

Res. 2016, Nov 29. pii: gkw1093. [Epub ahead of print]

11. Sloan-Heggen CM, Bierer AO, Shearer AE, Kolbe DL, Nishimura CJ, Frees KL, Ephraim SS, Shibata SB, Booth KT, Campbell CA, Ranum PT, Weaver AE, Black-Ziegelbein EA, Wang D, Azaiez H, Smith RJ. Comprehensive genetic testing in the clinical evaluation of 1119 patients with hearing loss. Hum Genet,135(4):441-50, 2016.

E N D O W E D C O R E FAC I L I T I E S

25


PROTEOMICS CORE FACILITY The mission of the Proteomics Core Facility

R. MARSHALL POPE, PHD

(PCF) is to provide mass-spectrometry

DIRECTOR

techniques to identify and quantify proteins and how they may be altered or regulated by post-translational modifications. Progress was made on two fronts. One was full implementation of the new Orbitrap LUMOS Mass spectrometer (ThermoFisher), awarded to Iowa through an HHMI technology grant (K. Campbell, PI). The second was implementing new bioinformatic workflow and analysis techniques aimed at providing streamlined quantitative proteomics analysis.

The LUMOS has millisecond sequencing speed and unmatched sensitivity. Most importantly, the LUMOS has measure masses with extreme accuracy while also having the ability to fragment peptides in a number of ways in parallel. These qualities combined allow the LUMOS to perform very well at identifying proteins in complex samples with high confidence. In addition, it also allows for different types of quantitation of proteins as well. With the LUMOS, UI researchers were able to1) track dysfunctional protein modifications in mouse heart mitochondria, 2) study exosomes secreted from human macrophages with and without parasite infection, 3) validate new technology to characterize targets of ubiquitin E3 ligases and 4) and perform structure-function studies on ion channels.

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An example of quantitative proteomics put into practice was work with Dr. Piper. They study the role of ubiquitin, which is a peptide that is covalently attached to a wide variety of substrates that in turn elicits a variety of responses. Covalent attachment of Ubiquitin to proteins is accomplished by a large array of Ubiquitin E3 ligase complexes (~240) and determines the fate for most proteins.. The Piper lab invented a way to create specific ‘reversal’ enzymes that inhibit ubiquitination events in dominant-negative way. They then purified all the ubiquitinated proteins from control and experimental samples to find specific proteins that were affected upon expression of a particular ‘reversal’ enzyme. Samples were then differentially labled using a method that the facility published last year and that we reviewed this year (3). We then implemented a new data processing program, MassQuant, that can then take complete spectra and calculate relative levels of each peptide and thus each protein between the differentially labeled samples. This work was recently submitted as a revised manuscript describing an extensive set of methods and tools for researchers interested in the function of any ubiquitin ligase (1). Going forward, the facility can now perform all aspects of sample processing, data collection, and data analysis to enable researchers to compare proteins, peptides, and post-translational modifications amongst several different samples allowing better protein discovery experiments and also enabling global analysis of changes in whole cell proteomes, signal transduction pathways, or protein complex formation. We also used the LUMOS and a different set of new bioinformatics tools to monitor biochemical changes within ion channels that could lead to a novel treatment for cystic fibrosis. In The PCF worked with the McCray and Ahern labs to suppress mutations in the CFTR gene by manipulating the protein translation machinery to ignore these mutations. In particular, we helped validate a novel tRNA engineering approach where tRNA sequences are customized to no longer recognize and suppress their cognate DNA codon sequences, but instead guide tRNA (and a cognate amino acid) to the disease missense codon, thus allowing the mutant gene to produce a normal protein. We implemented a proteomics approach to show the specificity of for how the engineered tRNA rescued CFTR activity. This approach allowed us to detect of 80% of the CFTR protein and definitively show the correct amino acid substitution in the sole targeted site of the mutant CFTR gene, representing the first successful and specific rescue of a missense stop mutation in CFTR.

E N D O W E D C O R E FAC I L I T I E S

27


Future The new LUMOS mass spectrometer and the recent implementation of new qualitative and quantitative data analysis tools has provided a strong infrastructure that can support a wide range of proteomic projects. We look forward to running more challenging projects like these and applying the differential comparisons in more novel applications. Currently, the LUMOS is already running near capacity and we hope to find funds to purchase a complementary instrument that can not only enhance the repertoire of analyses we can perform but also make it possible to retire 3 older instrument and consolidate costs. Recently, we moved our instrumentation to temporary space to allow the CCOM to renovate the proteomics core lab. This new design features a purposebuilt instrumentation room specifically engineered to support these sensitive pieces of equipment. It also features separate lab space and clean rooms for sample preparation. Finally, it features a data processing/analysis hub where investigators and facility personnel can perform and discuss data analysis and experimental protocols. Routine projects: • The PCF completed 2334 MALDI analyses for 32 investigators across 16 departments. • The PCF also completed 1116 LCMS projects for protein identification or quantification.

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January 1, 2016 — December 31, 2016 FINANCIAL SUMMARY

PROTEOMICS CORE FACILITY

Balance Forward Revenue Disbursements to UI Total Revenue

($35,273) Equipment Maintenance (9.8%)

Other (2.2%)

$145,746

Expenses Personnel $88,778 Equipment Lab Supplies $469 4.6% Equipment $4,862 Equipment Maintenance $10,437 Other $2,306 Lab Supplies (0.4%) Total Expenses $106,851 Ending Balance

Personnel (83.1%)

$3,621

PUBLICATIONS

PATENTS

1. Chris MacDonald, Stanley Winistorfer, R. Marshall Pope, Michael

1. Yu, Chi-Li and Pope, Robert: SAMPLING SYSTEMS,

E. Wright, and Robert C. Piper, (2017) Enzyme reversal to explore the

DEVICES, AND METHODS FOR ANALYZING SAMPLES,

function of yeast E3 ubiquitin-ligases, re-submitted to Traffic Jan 28, 2017.

US Provisional Patent Application No. 62/329,715 filed through the University of Iowa Research Foundation

2. John D. Lueck, Daniel T. Infield, Adam L. Mackey, R. Marshall Pope, Paul B. McCray and Christopher A. Ahern, (2016) Engineered tRNA

2. Li, Yalan and Pope, Robert: ACCELERATED SILVER

suppression of a CFTR nonsense mutation, preprint on bioRxiv, Nov

STAINING COMPATIBLE WITH MASS SPECTROMETRY,

20 2016, http://biorxiv.org/content/early/2016/11/20/088690.

patent application in preparation, Ballard Spahr LLP, Atlanta, GA. Attorney Docket No. 21087.0095U1

3. Yu, Brooks, Li, Subramanian, Summers, Pope, Rapid Proteomics to Prospect and Validate Novel Bacterial Metabolism Induced by Environmental Burden. In Arun K. Shukla, editor: Proteomics in Biology, Part B, Vol 586, MIE, UK: Academic Press, 2017, pp. 379-411.

PATENTS

E N D O W E D C O R E FAC I L I T I E S

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TISSUE PROCUREMENT CORE The mission of the Tissue Procurement

C. MICHAEL KNUDSON, MD, PHD CLINICAL ASSOCIATE PROFESSOR, DEPARTMENT OF PATHOLOGY—TRANSFUSION MEDICINE CLINICAL PROFESSOR, DEPARTMENT OF RADIATION ONCOLOGY

Core (TPC) is to facilitate basic and clinical biomedical research through the collection, storage, and distribution of human blood, human tissue, and tissue derivatives while assuring patient care and confidentiality are not compromised. The TPC currently promotes this function by supporting investigator projects directly and by providing collection and distribution services to various tissue specific repositories within the Carver College of Medicine and the Holden Comprehensive Cancer Center. Funds from the Roy J. Carver Charitable Trust were used in 2016 to support TPC personnel, primarily to allow the TPC to maintain sufficient staff and equipment to keep up with the core’s continued increase in utilization and expansion of services over the past few years. TPC Support for the Carver College of Medicine Research Enterprise Working in collaboration with the Holden Comprehensive Cancer Center (HCCC) and the Carver College of Medicine, the TPC functions as a key player in the federated biorepository model at the University of Iowa. In this model, the TPC is responsible for collecting, cryopreserving, and distributing biospecimens for research purposes. Other participants in the federated biorepository model include

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Figure 1. Venn Diagram representing the intersections of the various components of the federation of biorepositories. Figure 2. As a first step investigators should consult with TPC about biospecimen requirements for a research project. If frozen or fresh tissue, blood or a derivative from frozen tissue or blood is required they will generally work with TPC and MERs to obtain the required specimens. If the project can be addressed with FFPE blocks (TMA or IIHC slides for instance) they are generally referrred to the Population Research Core and specimens can be obtained from the Path Archives or VTR/LTR.

tissue-specific repositories (MERs) that focus on a variety

As reported last year the TPC was a significant player

of tumors including Breast, Colon, Pancreas, Melanoma,

in the selection, acquisition, and implementation of a

Sarcoma, Lymphoma, Renal and Bladder malignancies.

software platform (Labmatrix from Biofortis) and the

The TPC also collaborates with another major research

accompanying service (UIBioshare) to support the

initiative—the neuroendocrine tumor (NET) SPORE

federated biorepositories. The TPC works in collaboration

repository. TPC is currently working to help develop the

with UIBioshare to triage requests from investigators

Peds Oncology Bank in conjunction with researchers and

to funnel requests to the most appropriate repositories.

physicians from the UI Stead Family Children’s Hospital

The triaging scheme is shown in Figure 2.

Figure 1. Thus, by supporting the TPC, the Carver Trust indirectly supports each of these federated repositories.

In addition to coordinating with the MERs in the HCCC, the TPC has now fully integrated the services initially

E N D O W E D C O R E FAC I L I T I E S

31


offered by the UI Biobank to isolate DNA from blood

distribute tissue is significantly increased. Also the demands

and saliva for the purpose of supporting genetic studies.

have significantly shifted from whole tissues to the realm of

The UI Biobank consent is open-ended and allows

molecular biomaterials (DNA, RNA and protein). Addition-

investigators to contact participants later to obtain outcome

al services have been added, including in-facility specimen

information as well as information that might not be

sectioning, high quality nucleic acid extraction, and enhanced

available in the medical record. The incorporation of the

prospective collections. The expanded services along with ad-

UIBB DNA core into the TPC significantly expanded the

ditional outreach efforts have improved utilization of the TPC.

DNA processing capability of the TPC and brought a

Utilization has significantly increased over the last five years

number of additional investigators and projects under the

as demonstrated by the number of Carver College of Medi-

TPC umbrella. See TPC Utilization for more details.

cine investigators served Figure 3 and specimens distributed

TPC Utilization One of the goals of the TPC has been to grow utilization. With the rapidly evolving research environment the demand for biomaterials has shifted away from archived frozen tissue from a wide array of situations toward procurement of fresh specimens that meet very specific requirements. The shift to prospectively collected fresh specimens results in fewer numbers of specimens to be distributed but the effort to identify appropriate patients, obtain consent, and acquire and

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Figure 4. Support from the Carver Trust has allowed the TPC to: have sufficient staff and equipment to keep up with the increase in utilization over time; to adapt to the changing requirement for fresh biospecimens; and to participate in special projects described below. The slight dip in specimen distribution in 2016 largely reflects the completion of one project which used large numbers of individual frozen sections. The contribution

2 0 1 7 A N N U A L R E P O R T TO T H E C A R V E R T R U S T


January 1, 2016 — December 31, 2016 FINANCIAL SUMMARY

TISSUE PROCUREMENT CORE Balance Forward

($5,947)

Revenue Disbursements to UI Total Revenue

$138,100

Other (7.7%) Equipment Maintenance (4.8%)

Expenses Personnel $93,493 Equipment Lab Supplies $6,140 (0.7%) Travel $2,808 Equipment $874 Equipment Maintenance $5,687 Travel Other $9,125 (2.4%) Lab Supplies Total Expenses $118,127

Personnel (79.1%)

(5.2%)

Ending Balance

$14,025

of the DNA processing core over the last two years is shown in

Another service the TPC has added is the ability to isolate

Figures 5 and 6.

RNA from blood specimens. This service was added to support a clinical trial by a Carver College of Medicine

New Initiatives The TPC/Federated system collaborated with the HCCC to apply for a supplemental cancer center grant to provide fresh tissue to the National Cancer Institute (NCI) for the production of patient derived xenographs (PDX), a special type of human cell culture maintained in an animal, usually a mouse. This project is a nation-wide collaboration across a number of institutions and requires efficient collection and distribution in order to be successful. The UI TPC was the first center to be certified for collections and the first center to complete the initial contract with this program. Because of the successful partnership with the TPC, the NCI extended and expanded the supplement not once but three times and the possibility exists for further extension. The PDXs and cell lines that are created by the NCI from tissues and blood collected by the TPC will be used to form a national repository of readily available biological material to advance the understanding and treatment of a variety of cancers. Our experience with this project has allowed us to support a number of Carver College of Medicine researchers who are

investigator in the area of multiple sclerosis. The addition of this service positions the TPC to support other projects by Carver College of Medicine investigators. Conclusions and Future Directions The role of the TPC in supporting biomedical research within the Carver College of Medicine and the HCCC has changed substantially over the last several years. The TPC has increased its visibility and developed a reputation for providing high quality services at a very reasonable cost. As shown above the result has been a substantial increase in utilization and the TPC has been able to participate in key national initiatives and clinical trials. The TPC and the Federated repository system plan to continue to make affordable, high-quality human tissues available to all UI investigators performing high-quality research and the service is looking to expand the horizon outside the institutional boundaries. We believe that use of these biospecimens will facilitate the publication of high impact manuscripts and implementation of “personalized” medicine at UI Hospitals and Clinics and beyond.

also creating PDX’s with fresh tissues provided by the TPC.

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ENDOWED CHAIRS AND PROFESSORSHIPS

35


Dr. Charles Brenner’s research focuses on how different environmental and

CHARLES M. BRENNER, PHD

nutritional conditions lead to changes in an organism’s DNA and proteins. He discovered nicotinamide riboside (NR) as a vitamin and conducts experiments to determine how NR boosts metabolism in rodents and in people.

ROY J. CARVER CHAIR IN BIOCHEMISTRY CHAIR AND DEPARTMENT EXECUTIVE OFFICER, DEPARTMENT OF BIOCHEMISTRY

In 2016, funds from the Roy J. Carver Charitable Trust were used to perform the Brenner laboratory’s first animal experiments with NR and to perform the first clinical trial of NR, which showed that NR safely elevates nicotinamide adenine dinucleotide (NAD) metabolism in people.

PROFESSOR, DEPARTMENT OF BIOCHEMISTRY PROFESSOR, DEPARTMENT OF INTERNAL MEDICINE

Twelve years ago, in the course of doing experiments in yeast, Dr. Brenner’s research group discovered NR as a vitamin precursor of NAD, the central coenzyme of metabolism. Remarkably, this vitamin is now in common use by people: an estimated 200,000 US citizens take NR on a daily basis. NAD is required for human beings to convert fuels such as fats, proteins and carbohydrates into energy; to detoxify reactive oxygen species; to repair DNA; and to build and maintain healthy skin and musculature. Because NAD and each of these processes decline in aging, boosting NAD with NR has emerged as an approach to maintain optimum health. NR technologies have been licensed and commercialized. The main challenge in the science of NR is to determine the types of diseases and conditions that involve the greatest dysregulation of NAD that can be prevented or corrected by NR. To determine how NAD metabolism changes as a function of aging and disease, the Brenner laboratory developed quantitative targeted NAD metabolomics, a method to determine the levels of the full set of NAD metabolites in a tissue, blood sample, or food source. In 2016, targeted metabolomic technologies were used to determine the levels of NAD metabolites including NR in cow’s milk; to determine the effect of obesity and type 2 diabetes on NAD metabolism; to determine if NR is orally available to people; and to determine how a mouse with a gene deletion in the NR utilization pathway would behave.

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

CHARLES M BRENNER, PHD Balance Forward

($18,443)

Revenue Disbursements to UI Total Revenue

Equipment Maintenance (0.4%)

$97,282 $97,282 Personnel (99.6%)

Expenses Personnel $131,442 Other $502 Total Expenses $131,944 Ending Balance

($53,104)

The publication in the Journal of Nutrition was important

Investigaciones Oncologicas. In December, he was the keynote

because it established that NR is a natural product that is

speaker at Des Moines University Research Day.

present in our food supply. The publication in Scientific Reports was important because it showed that NR protects overfed and type 2 diabetic mice from fatty liver; aids in

SELECTED PUBLICATIONS

glycemic control; and also provides almost complete protection from prediabetic and type 2 diabetic neuropathy in mice. These data are being used to design human clinical trials to test the activity of NR in diabetic and chemotherapeutic neuropathy. Indeed, one such trial to test NR as a protective agent against paclitaxel-induced peripheral neuropathy in breast cancer patients has already been funded by the National Cancer Institute (Donna Hammond, PI). The first article in Nature Communications showed that NR safely boosts NAD metabolism in people. In the course of doing this work, the Brenner laboratory discovered a highly sensitive biomarker of boosting NAD. The

1. Trammell SA, Yu L, Redpath P, Migaud ME, Brenner C. Nicotinamide Riboside Is a Major NAD+ Precursor Vitamin in Cow Milk. J Nutr. 2016 May 146(5):957-63. doi: 10.3945/jn.116.230078.

2. Trammell SA, Weidemann BJ, Chadda A, Yorek MS, Holmes A, Coppey LJ, Obrosov A, Kardon RH, Yorek MA, Brenner C. Nicotinamide Riboside Opposes Type 2 Diabetes and Neuropathy in Mice. Sci Rep. 2016 May 27;6:26933. doi: 10.1038/srep26933.

3. Trammell SA, Schmidt MS, Weidemann BJ, Redpath P, Jaksch F,

second article in Nature Communications showed that a

Dellinger RW, Li Z, Abel ED, Migaud ME, Brenner C. Nicotinamide

mouse with a gene deletion in nmrk1 cannot use NR or

riboside is uniquely and orally bioavailable in mice and humans. Nat

a related compound, NMN, to boost NAD metabolism.

Commun. 2016 Oct 10;7:12948. doi: 10.1038/ncomms12948.

Work performed in 2016 will enable further mechanistic and translational experiments with NR in 2017.

4. Ratajczak J, Joffraud M, Trammell SA, Ras R, Canela N, Boutant M, Kulkarni SS, Rodrigues M, Redpath P, Migaud ME, Auwerx J, Yanes O,

During 2016, Dr. Brenner was honored with two major

Brenner C. CantĂł C. NRK1 controls nicotinamide mononucleotide

lectureships. In October, he was a distinguished speaker

and nicotinamide riboside metabolism in mammalian cells. Nat

at the Spanish cancer center in Madrid, Centro Nacional

Commun. 2016 Oct 11;7:13103. doi: 10.1038/ncomms13103.

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KEVIN P. CAMPBELL, PHD Dr. Kevin Campbell’s research is aimed at:

ROY J. CARVER BIOMEDICAL RESEARCH CHAIR IN MOLECULAR

identifying and defining mechanisms that

PHYSIOLOGY AND BIOPHYSICS

generating new knowledge regarding

CHAIR AND DEPARTMENT EXECUTIVE OFFICER,

the function of the protein dystroglycan,

DEPARTMENT OF MOLECULAR PHYSIOLOGY AND BIOPHYSICS PROFESSOR , DEPARTMENT OF MOLECULAR

cause various forms of muscular dystrophy;

PHYSIOLOGY AND BIOPHYSICS

which is an important component of muscle and defective in several forms of muscular dystrophy; and developing

PROFESSOR, DEPARTMENT OF NEUROLOGY

therapeutic strategies for the treatment

PROFESSOR, DEPARTMENT OF INTERNAL MEDICINE

of these diseases. During the past year,

INVESTIGATOR, HOWARD HUGHES MEDICAL INSTITUTE

Dr. Campbell’s laboratory has focused on determining the structural basis of the interaction between dystroglycan in muscle and the underlying support structure known as the extracellular matrix, and how abnormalities in this interaction causes muscular dystrophy. Support from the Roy J. Carver Charitable Trust Endowed Chair has had a resounding impact on the research programs and activities in Dr. Campbell’s laboratory over the past year. The funds have been used to: support one postdoctoral fellow and several undergraduate research assistants; purchase supplies and maintain the colonies of mice that are essential for this group’s cutting-edge research toward discoveries in the field of muscular dystrophy; and support projects with the Carver College of Medicine NMR Core Facility that have greatly facilitated Dr. Campbell’s structural biology research. Like many other proteins, dystroglycan is modified by the addition of particular sugar molecules, and this “glycosylation” is required for it to function correctly. The proper assembly of sugars on this protein depends on the coordinated activities of a number of enzymes known as glycosyltransferases, which sequentially assemble a complex

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2 0 1 7 A N N U A L R E P O R T TO T H E C A R V E R T R U S T


array of sugar molecules on dystroglycan. This modification

Dr. Campbell was elected a Lifetime Achievement Fellow

of dystroglycan is essential for making it “sticky” so that it

by The American Society for Cell Biology (ASCB). These

can bind to components of the extracellular matrix. However,

Fellows are elected on the basis of a career of exceptional

stickiness is not sufficient for proper function; the sugars must

contributions to science and for their service to the ASCB.

be placed in a specific sequence and orientation. Indeed, errors in their placement can cause dystroglycan to lose its

Dr. Campbell was presented with the Greg Marzolf Jr.

ability to bind its partner proteins in the extracellular matrix,

Foundation Symposium Visiting Lecture Award at the

and this in turn causes congenital muscular dystrophies—

University of Minnesota. Founded in 2002 to commemorate

neuromuscular diseases characterized by a progressive

the life of Greg Marzolf Jr., the Foundation raises awareness

loss of muscle function. However, the basis of recognition

of muscular dystrophy through philanthropic activities, and

of this unusual carbohydrate structure by laminin and

supports research toward a cure for muscular dystrophy. The

other similar proteins remained a mystery until now.

Foundation is also dedicated to funding education and research programs that benefit children and families suffering from

Funding from the Carver Trust directly supported all of the

muscular dystrophy, and to providing support for patient care.

research projects in Dr. Campbell’s laboratory, which rely heavily on mouse models and laboratory supplies. In the past

In addition to heading his laboratory and serving as Chair

year Dr. Campbell’s research program was very successful,

and Department Executive Officer of the Department of

publishing three high-impact papers in Nature Chemical Biology,

Molecular Physiology and Biophysics, Dr. Campbell directs

eLife and Proceedings of the National Academy of Sciences. The

the Paul D. Wellstone Muscular Dystrophy Cooperative

study reported in Nature Chemical Biology (2016; 12(10):810-

Research Center (MDRC) at the Carver College of Medicine.

4) was particularly exciting. Entitled “Structural basis of

Established in 2005, this Center brings together researchers

laminin binding to the LARGE glycans on dystroglycan,” this

and clinicians to facilitate laboratory discoveries and

article reviewed the mechanism whereby a chain of sugars

translate them into clinical applications for the treatment

that is critical for proper function of dystroglycan in many

of congenital and limb-girdle muscular dystrophies.

tissues, especially muscle and brain, binds to the extracellular protein laminin. It reports on NMR binding studies and the analysis of crystal structures of key parts of laminin bound to a sugar chain that is synthesized by an enzyme known as LARGE. The results reveal that regions of laminin are Ca2+-dependent lectins that bind specifically to sugars known as glucuronic acid-xylose disaccharides. This mode of sugar-chain recognition is unprecedented, and explains the strength of binding measured by NMR. The reason this sugar group is so important is that, through dystroglycan, it links internal components of the cell to the extracellular matrix, a structure that fills the space between the cells. This matrix serves as a scaffold, holding the cells together. It is well established that the interaction of the cell with this component of its environment is crucial for tissue architecture, cell maturation, and cell migration. Thus, Dr. Campbell’s work provides the first atomic-level insights into the structure and function of a unique interaction between proteins and sugars that is perturbed in muscular dystrophies. Collectively, his results represent a major advance in our understanding of the mechanisms that underlie the dystroglycanopathies.

E N D OW E D C H A I R S A N D P RO F ESS O R S H I P S

39


PUBLICATIONS

1. de Greef J.C., Hamlyn R., Jensen B.S., O’Campo Landa, R., Levy,

7. Briggs, D., Yoshida-Moriguchi, T., Zheng, T., Venzke, D., Anderson,

J.R., Kobuke, K., Campbell, K.P. Collagen VI Deficiency Improves

M., Strazzulli, A., Moracci, M., Yu, L., Hohenester, E., Campbell,

Muscle Pathology, but Not Muscle Function, in the Y-Sarcoglycan-Null

K.P. Structural Basis of Laminin Binding to the LARGE Glycans on

Mouse. Human Molecular Genetics 2016; 25(7):1357-1369. doi:10.1093/

Dystroglycan. Nature Chemical Biology 2016; 12(10):810-814. doi:10.1038/

hmg/ddw018 PMID: 26908621 PMCID: PMC4787905

nchembio.2146 PMID: 27526028 PMCID: PMC5030134

2. Agre, P., Bertozzi, C., Bissell, M, Campbell, K.P., Cummings, R.,

8. Inamori, K., Beedle, A.M., Beltrán-Valero de Bernabé, D., Wright, M.E.,

Desai, U., Estes, M., Flotte, T., Fogleman, G., Gage, F., Ginsburg, D.,

Campbell, K.P. LARGE2-dependent Glycosylation Confers Laminin-

Gordon, J., Hart, G., Hascall, V., Kiessling, L., Kornfeld, S., Lowe, J.,

Binding Ability on Proteoglycans. Glycobiology 2016, 26(12): 1284-1296.

Magnani, J., Mahal, L., Medzhitov, R., Roberts, R., Sackstein, R., Sarker,

doi: 10.1093/glycob/cww075. PMID: 27496765 PMCID: PMC5137251

R., Schnaar, R., Schwartz, N., Varki, A., Weissman, I. Training the Next Generation of Biomedical Investigators in Glycosciences. Journal of Clinical Investigation 2016;126(2):405-408. doi:10.1172/JCI85905

9. Rader, E., Turk, R., Willer, T., Beltrán, D., Inamori, K., Peterson, T.A., Engle, J., Prouty, S., Matsumura, K., Saito, F., Anderson, M.E., Campbell, K.P. Role of dystroglycan in limiting contraction-induced

3. Blaeser A., Harper A., Campbell K.P., Lu Q. L. Report:

injury to the sarcomeric cytoskeleton of mature skeletal muscle. Proceedings

Fourth International Workshop for Glycosylation Defects

of the National Academy of Sciences U S A 2016 Sep 27;113(39):10992-7.

in Muscular Dystrophies. Journal of Genetic Syndromes & Gene

doi:10.1073/pnas.1605265113 PMID: 27625424 PMCID: PMC5047148

Therapy 2016, 7:1. doi:10.4172/2157-7412.1000286 10. Frϋh, S., Romanos, J., Panzanelli, P., Bϋrgisser, D., Tyagaragan, 4. Allamand, V., Bonaldo, P., Bӧnnemann, C., Brown, S.C., Burkin,

S.K., Campbell, K.P., Santello, M., Fritschy, JM. Neuronal

D., Campbell, K.P., Durbeej-Hjalt, M., Girgenrath, M., Nevo,Y.,

Dystroglycan is Necessary for Formation and Maintenance of

Ruegg, M., Saunier, M., Toda, T., Willmann, R. 212th ENMC

Functional CCK-Positive Basket Cell Terminals on Pyramidal Cells.

workshop: Animal models of Congenital Muscular Dystrophies,

The Journal of Neuroscience 5 October 2016, 36(40): 10296-10313.

Naarden, The Netherlands, May 29th-31st, 2015. Neuromuscular

doi:10.1523/JNEUROSCI.1823-16.2016 PMID: 27707967

Disorders 2016; 26(3): 252-259. doi:10.1016/j.nmd.2016.02.002 11. Jerber, J., Zaki, M.S., Al-Aama, J.Y., Ozgur Rosti, R., Ben-Omran, 5. Praissman, J.L., Willer, T., Sheikh, M. O., Toi, A, Chitayat, D, Lin, Y,

T., Dikoglu, E., Silhavy, J. L., Caglar, C., Musaev, D., Albrecht, B.,

Lee, H, Stalnkaker, S, Wang, S, Prabhakar, P, Nelson, SF, Stemple, D.L.,

Campbell, K.P., Willer, T., Almuriekhi, M., Okay Çağlayan, A., Vajsar,

Moore, S.A., Moremen, K.W., Campbell, K.P., Wells, L. The Functional

J., Bilgϋvar, K., Ogur, G., Abou Jamra, R., Gϋnel, M., Gleeson, J.G.

O-Mannose Glycan on α-Dystroglycan Contains a Phospho-ribitol Primed

Biallelic Mutations in TMTC3, Encoding a Transmembrane and TPR-

for Matriglycan Addition. eLife 2016, 29;5:e14473. doi:http://dx.doi.

Containing Protein, Lead to Cobblestone Lissencephaly. American Journal

org/10.7554/eLife.14473 PMID: 27130732 PMCID: PMC4924997

of Human Genetics 3 November 2016; 99(5): 1181–1189. doi:10.1016/j. ajhg.2016.09.007 PMID: 27773428 PMCID: PMC5097947

6. Turk R., Hsiao J.J., Smits, M.M., Ng, B.H., Pospisil, T.C., Jones, K.S., Campbell, K.P., Wright, M.E. Molecular Dissection of

12. Zhu, Q., Venzke, D., Walimbe, A.S., Anderson, M.E., Fu, Q.,

Membrane Protein Complexes Associated with Muscular Dystrophy.

Kinch, L., Wang,W., Chen, X., Grishin, N., Huang, N., Yu, L.,

Molecular Cell Proteomics 2016 Jun;15(6):2169-85. doi:10.1074/

Dixon, J.E., Campbell, K.P., Xiao, J. Structure of Protein

mcp.M116.059188 PMID: 27099343 PMCID: PMC5083101

O-Mannose Kinase Reveals a Unique Active Site Architecture. eLife 2016;10.7554/eLife.22238. doi:http://dx.doi.org/10.7554/ eLife.22238 PMID: 27879205 PMCID: PMC5142810

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

KEVIN P. CAMPBELL, PHD Balance Forward Revenue Disbursements to UI Total Revenue Expenses Lab Supplies Total Expenses Ending Balance

($80,215)

Lab Supplies (100%)

$69,301

$61,756 $61,756 ($72,670)

E N D OW E D C H A I R S A N D P RO F ESS O R S H I P S

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JOHN F. ENGELHARDT, PHD Dr. John Engelhardt’s research focuses on developing gene therapies for inherited and

ROY J. CARVER CHAIR IN MOLECULAR MEDICINE CHAIR AND DEPARTMENT EXECUTIVE OFFICER,

environmentally induced diseases. Included are two major fields of research emphasis:

1) the study of lung molecular and cellular

DEPARTMENT OF ANATOMY AND CELL BIOLOGY

biology as it relates to cystic fibrosis lung

DIRECTOR, CENTER FOR CYSTIC FIBROSIS AND

disease and treatments using gene therapy;

OTHER GENETIC DISEASES PROFESSOR, DEPARTMENT OF ANATOMY AND CELL BIOLOGY PROFESSOR, DEPARTMENT OF INTERNAL MEDICINE

and 2) the pathogenesis and treatment of cystic fibrosis related diabetes through studies in animal models and humans.

Funds from the Roy J. Carver Charitable Trust were used in 2016 for research projects studying: 1) stem cell responses to injury in the lung and pancreas as they relate to cystic fibrosis disease processes; and 2) the development of improved viral vectors for gene therapy of cystic fibrosis lung disease. Identification of novel stem cell compartments in airway submucosal glands: Submucosal glands in the airway have been hypothesized to contain stems cells that assist in repairing the airways following injury. Submucosal glands also express the highest levels of cystic fibrosis transmembrane conductance regulator (CFTR) protein in the lung—the protein that is defective in cystic fibrosis (CF). We have previously shown that submucosal glands have defective secretions in CF and altered regenerative responses following airway injury. During the past year, we discovered a link between the biology of primordial glandular stem cells (i.e., those that give rise to submucosal glands during development) and stem cells that reside in adult submucosal glands. Both primordial glandular stem cells and adult glandular stem cells utilize secreted molecules called Wnts to facilitate stem cell expansion during development and following injury. We discovered that components of the glandular stem cell niche (i.e., the neighborhood where stem cells reside) are localized to regions of submucosal glands

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where specific Wnt-active supporting cells reside. Isolation

humanized virus called AV2.5T generated by Dr. Zabner.

of these adult glandular stem cells demonstrated that they

Interestingly, this virus infects cultured differentiated human

have a unique capacity to differentiate into all components of

airway epithelia very well, but does not infect the pig airway.

the airway epithelium and have a greater capacity to expand

We tested AV2.5T in ferrets and find that like humans, the

in culture. These findings may assist in the development of

virus infects the ferret airway very well. We are completing

stem cell therapies for cystic fibrosis and other lung diseases.

the preclinical testing of AV2.5T virus in ferrets and are planning to move the recombinant virus into a clinical trial

Mechanism of Diabetes in Cystic Fibrosis: Cystic fibrosis related

for CF lung disease. A second area of growth has involved

diabetes (CFRD) is the most common complication of cystic

a collaborative effort between the Engelhardt group and

fibrosis (CF) and is associated with increased morbidity

Pfizer to evaluate rAAV/HBoV1 chimeric virus for gene

and mortality. At this time last year, Dr. Engelhardt and his

therapy of CF lung disease. Studies over the past year have

collaborators on an NIH-funded R24 Team Science grant had

demonstrated rAAV/HBoV1 virus can be repetitively given

uncovered a novel mechanism through which the CF pancreas

to the lungs of ferrets without an immune response that

rebuilds islets following exocrine pancreas destruction. These

prevents second infection. An advantage of the rAAV/HBoV1

studies utilized a novel CF ferret model. More recently, they

virus is that the capsid is large enough to package the entire

obtained evidence that new beta cells (i.e., the cells that

CFTR cDNA with a strong promoter, unlike rAAV vectors

secrete insulin to regulate blood glucose) form during a stage

that require a CFTR minigene due to their smaller size. The

of pancreatic remodeling associated with improved glucose

collaboration with Pfizer has the goal to move this vector

tolerance. Evidence that beta cell precursors replicate during

forward to a CF lung clinical trial over the next three years.

this stage is not only important for therapies targeting CFRD, but also other forms of diabetes. During the last year, this team

During the past year, Dr. Engelhardt has remained

has also demonstrated that similar alterations to the pancreas

administratively active as Chair of the Department of

likely occur in very young children with CF. This clinical trial

Anatomy and Cell Biology. In this context, he recently

in CF children 3 months to 6 years of age, has demonstrated

recruited Dr. Sam Young from the Max Plank Institute. Dr.

that a significant portion of children 2 to 4 years of age have

Young’s research focuses on the mechanisms of synaptic

abnormal glucose tolerance. Importantly, by the time these

transmission and neuronal circuitry and their dysfunctional

children are 5 to 6 years of age, glucose tolerance improves

in hearing loss. His group is also developing gene therapies

in a manner similar to CF ferrets. These findings suggest that

for hearing loss. He will reside within the Auditory

CF young children can likely also regenerate islets following

Neuroscience Group of the new Neuroscience Institute. Dr.

early inflammation that destroys the exocrine pancreas.

Engelhardt also participated in strategic planning for research

Current efforts are targeted at identifying the islet stem

programs within the CCOM and will continue playing a

cell precursors and understanding the biology that controls

role in small groups as the strategic plan is implemented.

this regenerative process. Such efforts may lead to novel

His research has been productive over the past year with

approaches for treating CFRD and other forms of diabetes.

15 manuscripts, three new patent applications, and the development of new sponsored research with Pfizer. Dr.

Gene Therapy for CF Lung Disease. The Engelhardt laboratory

Engelhardt also continues to mentor a large number of

has a long-standing commitment to the development of gene

students including five graduate students and five post-docs.

therapy for CF lung disease. In this context, they have been working with two types of viral vectors: recombinant adenoassociated viral (rAAV) and a recently engineered chimeric virus composed of rAAV genome and human Boca Virus (HBoV1) capsid (called rAAV/HBoV1). At this time last year, they had solved some major issues with rAAV vector design and during the present year have been working to test rAAV vectors in CF ferrets and pigs with collaborators (Welsh, Zabner, and McCray). Their lead rAAV vector includes a

E N D OW E D C H A I R S A N D P RO F ESS O R S H I P S

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PUBLICATIONS

1. Deng, X., Yan, Z., Cheng, F., Engelhardt, J.F., and Qiu, J. 2016. Replication of an Autonomous Human Parvovirus in Non-dividing Human Airway Epithelium Is Facilitated through the DNA Damage and Repair Pathways. PLoS Pathog 12:e1005399. pmcid:PMC4713420.

2. Evans, T.I., Joo, N.S., Keiser, N.W., Yan, Z., Tyler, S.R., Xie, W., Zhang, Y., Hsiao, J.J., Cho, H.J., Wright, M.E., Wine, J.J., and Engelhardt, J.F. 2016. Glandular Proteome Identifies Antiprotease Cystatin C as a Critical Modulator of Airway Hydration and Clearance. Am J Respir Cell Mol Biol 54:469-481. pmcid:PMC4821051.

3. Gibson-Corley, K.N., Meyerholz, D.K., and Engelhardt, J.F. 2016. Pancreatic pathophysiology in cystic fibrosis. J Pathol 238:311-320. pmcid:PMC4699289.

4. Hackett, T.L., Ferrante, S.C., Hoptay, C.E., Engelhardt, J.F., Ingram, J.L., Zhang, Y., Alcala, S.E., Shaheen, F., Matz, E., Pillai, D.K., and Freishtat, R.J. 2016. A Heterotopic Xenograft Model of Human Airways for Investigating Fibrosis in Asthma. Am J Respir Cell Mol Biol. (In Press)

5. Lynch, T.J., Anderson, P.J., Xie, W., Crooke, A.K., Liu, X., Tyler, S.R., Luo, M., Kusner, D.M., Zhang, Y., Neff, T., Burnette, D.C., Walters, K.S., Goodheart, M.J., Parekh, K.R., and Engelhardt, J.F. 2016. Wnt Signaling Regulates Airway Epithelial Stem Cells in Adult Murine Submucosal Glands. Stem Cells. (In Press)

6. Mou, H., Vinarsky, V., Tata, P.R., Brazauskas, K., Choi, S.H., Crooke, A.K., Zhang, B., Solomon, G.M., Turner, B., Bihler, H., Harrington, J., Lapey, A., Channick, C., Keyes, C., Freund, A., Artandi, S., Mense, M., Rowe, S., Engelhardt, J.F., Hsu, Y.C., and Rajagopal, J. 2016. Dual SMAD Signaling Inhibition Enables Long-Term Expansion of Diverse Epithelial Basal Cells. Cell Stem Cell 19:217-231. pmcid:PMC4975684.

7. Shen, W., Deng, X., Zou, W., Engelhardt, J.F., Yan, Z., and Qiu, J. 2016. Analysis of cis and trans Requirements for DNA Replication at the Right-End Hairpin of the Human Bocavirus 1 Genome. J Virol 90:7761-7777. pmcid:PMC4988151.

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

JOHN ENGELHARDT, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$167,870

Other (19.8%)

$93,475

Expenses Personnel $51,435 Lab Supplies/Animals $34,919 Other $21,269 Total Expenses $107,623 Ending Balance

$153,723

8. Spencer, N.Y., Yan, Z., Cong, L., Zhang, Y., Engelhardt, J.F., and

Personnel (47.8%)

Lab Supplies/Animals (32.4%)

Century - Research Challenges and Opportunities: Summary of a

Stanton, R.C. 2016. Definitive localization of intracellular proteins: Novel

National Institute of Diabetes and Digestive and Kidney Diseases

approach using CRISPR-Cas9 genome editing, with glucose 6-phosphate

Workshop. Pancreas 45:1365-1375. pmcid:PMC5117429.

dehydrogenase as a model. Anal Biochem 494:55-67. pmcid:PMC4695245. 12. Xu, Q., Choksi, S., Qu, J., Jang, J., Choe, M., Banfi, B., Engelhardt, 9. Steines, B., Dickey, D.D., Bergen, J., Excoffon, K.J., Weinstein, J.R., Li, X., Yan, Z., Alaiwa, M.H., Shah, V.S., Bouzek, D.C.,

J.F., and Liu, Z.G. 2016. NADPH Oxidases Are Essential for Macrophage Differentiation. J Biol Chem 291:20030-20041. pmcid:PMC5025689.

Powers, L.S., Gansemer, N.D., Ostedgaard, L.S., Engelhardt, J.F., Stoltz, D.A., Welsh, M.J., Sinn, P.L., Schaffer, D.V., and Zabner, J.

13. Yi, Y., Norris, A.W., Wang, K., Sun, X., Uc, A., Moran, A.,

2016. CFTR gene transfer with AAV improves early cystic fibrosis

Engelhardt, J.F., and Ode, K.L. 2016. Abnormal Glucose

pig phenotypes. JCI Insight 1:e88728. pmcid:PMC5033908.

Tolerance in Infants and Young Children with Cystic Fibrosis. Am J Respir Crit Care Med 194:974-980. pmcid:PMC5067820.

10. Sun, Z., Yu, W., Sanz Navarro, M., Sweat, M., Eliason, S., Sharp, T., Liu, H., Seidel, K., Zhang, L., Moreno, M., Lynch, T., Holton, N.E.,

14. Yi, Y., Sun, X., Gibson-Corley, K., Xie, W., Liang, B., He, N., Tyler,

Rogers, L., Neff, T., Goodheart, M.J., Michon, F., Klein, O.D., Chai,

S.R., Uc, A., Philipson, L.H., Wang, K., Hara, M., Ode, K.L., Norris,

Y., Dupuy, A., Engelhardt, J.F., Chen, Z., and Amendt, B.A. 2016.

A.W., and Engelhardt, J.F. 2016. A Transient Metabolic Recovery from

Sox2 and Lef-1 interact with Pitx2 to regulate incisor development and

Early Life Glucose Intolerance in Cystic Fibrosis Ferrets Occurs During

stem cell renewal. Development 143:4115-4126. pmcid:PMC5117215.

Pancreatic Remodeling. Endocrinology 157:1852-1865. pmcid:PMC4870869.

11. Uc, A., Andersen, D.K., Bellin, M.D., Bruce, J.I., Drewes, A.M.,

15. Zou, W., Cheng, F., Shen, W., Engelhardt, J.F., Yan, Z., and

Engelhardt, J.F., Forsmark, C.E., Lerch, M.M., Lowe, M.E.,

Qiu, J. 2016. Nonstructural Protein NP1 of Human Bocavirus

Neuschwander-Tetri, B.A., O’Keefe, S.J., Palermo, T.M., Pasricha,

1 Plays a Critical Role in the Expression of Viral Capsid

P., Saluja, A.K., Singh, V.K., Szigethy, E.M., Whitcomb, D.C.,

Proteins. J Virol 90:4658-4669. pmcid:PMC4836317.

Yadav, D., and Conwell, D.L. 2016. Chronic Pancreatitis in the 21st

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PAUL B. MCCRAY, JR., MD Dr. Paul McCray is a pediatric pulmonologist with long-standing interests in cystic

ROY J. CARVER CHAIR IN PULMONARY RESEARCH

fibrosis, epithelial biology, innate immunity, host-pathogen interactions, genomics,

INTERIM DIRECTOR, ALLERGY/PULLMONARY

DIVISION, DEPARTMENT OF PEDIATRICS EXECUTIVE VICE CHAIR, DEPARTMENT OF

PEDIATRICS PROFESSOR, DEPARTMENT OF PEDIATRICS PROFESSOR, DEPARTMENT OF MICROBIOLOGY

and the applications of gene therapy for inherited diseases. One goal of Dr. McCray’s work is to develop better treatments for genetic diseases affecting the pediatric population, through basic and translational research, teaching, and the training of new investigators. In 2016, support from the Carver Trust has helped support a new gene editing project in the lab. Funds from the Carver trust were used to further advance a gene editing project. The goal of this project is to apply CRISPR/Cas9 gene editing tools towards the correction of mutations in the CFTR gene, mutations that cause cystic fibrosis. GENE THERAPY FOR PEDIATRIC GENETIC DISEASES Dr. McCray’s laboratory continues to work to advance gene therapy to correct genetic diseases including cystic fibrosis and hemophilia A (a common bleeding disorder where patients are deficient in factor VIII, a protein required for blood clotting). He continues to work on the development of gene delivery vehicles (vectors) that can insert a copy of a therapeutic gene into the host cell DNA. This approach is called “gene addition”. If the therapeutic gene can be introduced into a population of cells that persist, the therapeutic correction could be long lasting. In these studies there are two different kinds of vectors, one called piggyBac and the other a lentivirus.

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Gene Editing as an approach to repair mutations

also being produced for testing. For the F508del mutation we

in the CFTR gene.

generated AAV2.5T and bocavirus vectors targeting the human

In 2015, Dr. McCray’s laboratory initiated a new high risk project to investigate a “gene editing” approach to addressing the problems caused by mutations in the CFTR gene that are causative for cystic fibrosis. The rapidly expanding CRISPRCas9 toolbox has increased access to gene editing technology and accelerated the pace of discovery. Dr. McCray’s focus is to advance the CRISPR-Cas9 system to modify the CFTR gene in somatic cells. The overall concept is that it is possible to target a population of cells in the airway with progenitor capacity and achieve persistent CFTR gene editing. The experiments underway will rigorously test the feasibility of CRISPR-Cas9 mediated gene editing in somatic cells and set the stage for future experiments in the CF pig model. In the past year, significant advances have occurred in building and testing the necessary tools for the studies proposed. To date we have begun to evaluate the efficiency of Streptococcus pyogenes (Sp) and Staphylococcus aureus (Sp) Cas9 in creating double strand breaks (DSBs) and insertions or deletions (indels) at specific DNA loci in model cells. Using human HEK293 cells and pig PK15 cells, we screened many guide RNAs for efficiency of creating DSBs. From these screens, we identified guide RNAs that introduce indels at a frequency of 30-50%. The guide RNAs with the best indel frequencies were subsequently moved into viral vectors for further testing in primary human cells. Identify the optimal delivery strategy to achieve efficient CFTR locus editing We are focusing on the use of adeno-associated virus (AAV), human bocavirus 1 (HBoV1), and adenovirus (Ad) to deliver the gene editing components. The AAV capsid 2.5T was engineered to efficiently enter human airway cells. We anticipate further optimizing delivery strategies for candidate CRISPR-Cas9, guide RNAs, and HR templates based on experiments using in vitro model systems. To date we have prepared AAV and bocavirus vectors to deliver gene editing reagents. The vectors were first tested in simple cell models. For the 3849+10 C>T mutation, we evaluated the AAV2.5T vector expressing both the 2 guide RNAs and the Sa Cas9 cDNA from a single vector. In HT1080 cells with an ~90 percent transduction efficiency, we observed an indel frequency of ~55 percent. The bocavirus vectors are

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mutation. The AAV2.5T delivery of guide RNA, Sp Cas9 cDNA, and a homologous recombination template requires 2 vectors. For bocavirus, using the Sa Cas9 cDNA, we deliver all 3 components a single vector. This is because the packaging capacity of the bocavirus vector is ~1 kb greater than that of a standard AAV vector. We evaluated the AAV2.5T vector in HT1080 cells and observed an indel frequency >50percent. We recently began further studies in primary welldifferentiated primary cultures of non-CF or CF human airway epithelia. Using these in vitro models, our goal is to determine the efficiency of CRISPR-Cas9 gene editing and the efficiency of HR. We will also investigate the efficacy of gene editing in terminally differentiated cells as well as progenitor cell types. We hypothesize that therapeutically relevant levels of gene editing can be achieved in airway epithelia in vitro. We first evaluated the vectors targeting the 3849+10 C>T mutation. We tested the efficacy of introducing DSBs in CFTR intron 22-23 Figure 1. At 7 and 28 days post transduction, indels were observed and the frequency increased over time. At 28 days post transduction, in 4 independent samples, we observed an indel frequency of 43 to 50percent. To further document the deletion of the deleterious sequence from intron 22-23, we performed additional studies as outlined in Figure 2. Following the vector treatment, the aberrant transcript was no longer detected. This result has been further validated using droplet digital PCR. To date we have only been able to evaluate this effect in cells from a single human donor. Our next goals include testing the vector in additional human CF cells and evaluating for functional correction of anion transport. Using 2 AAV2.5T vectors we have evaluated the efficiency of introducing DSBs at the F508del locus, and the efficiency of homologous recombination (HR). The cells evaluated were homozygous for the F508del mutation. As shown in Figure 3, following surveyor assay, sequencing of PCR amplicons, and RFLP, we observed an indel percentage of ~35percent and HR frequency of ~3 percent. This is not unexpected since HR is known to be especially inefficient in non-dividing cells. With this level of correction we do not expect to see functional correction. However, we are encouraged that we may be near the threshold for detecting functional correction. It is

47


interesting that the time required for introduction of indels

Here, we plan to utilize single or dual Staphylococcus pyogenes

in primary cells appears to be considerably slower than that

(Sp) Cas9- and/or Staphylococcus aureus (Sa) Cas9-specific

required in simple cell models. We plan to investigate several

single guide RNAs introduced by non-viral or viral delivery

strategies that might increase the HR frequency. These include

to remove and or replace the deleterious mutation and effect

evaluating alternative promoters for expression of the guide

repair by NHEJ or HDR in patient-derived conditionally

RNAs and the Cas9 gene and studies with additional vectors.

reprogrammed or primary airway epithelial cells. We tested the efficacy of different delivery mechanisms by assessing

Use CRISPR-Cas9 gene editing to correct the CF

the nature of indels, the indel efficiency, Cas9 expression,

defects in vitro.

vector genome copy numbers, vector persistence and HDR

By restoring CFTR-mediated anion transport, we have

efficiency in cell lines and primary airway epithelial cells.

the opportunity to repair defects in CF epithelia. For these studies we will ask: Will gene editing in airway epithelia correct the anion transport, ASL pH, and bacterial killing defects? In addition, we will identify the off-target effects of our optimized CFTR-targeted guide RNAs using a genomewide approach to map DSBs at nucleotide resolution using a method termed BLESS (direct in situ breaks labeling, enrichment on streptavidin and next-generation sequencing). We have not yet begun these studies.

Our results thus far demonstrate that both SpCas9 and SaCas9 genome editing can disrupt a mutation and/ or induce correction by HDR at different regions of CFTR locus. These early results suggest that CRISPR/Cas9 genome editing may have utility for the correction of patient-specific CFTR mutations. The availability of Carver Chair funds has been invaluable to Dr. McCray’s ability to develop and advance this

Figure 1. AAV2.5T vector transduction of conditionally reprogrammed human airway epithelial cells, compound heterozygous for F508del/3049+10 C>T. Cells were treated with AAV2.5T vector and evaluated by PCR and sequencing for deletions in intron 2223 at 7 and 28 days post transduction. At 28 days, in 4 independent samples, we observed an indel percentage from 43 to 50 percent indicating efficient deletion of the aberrant splice site.

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project. This support has allowed his lab to pursue new

director of the Allergy/Immunology and Pulmonary Division

areas of investigation, enhance existing programs, advance

in the Department of Pediatrics. He served the Department

high risk concepts, and seek additional extramural grant

of Pediatrics from 2007 to 2015 as Vice Chair for Research,

support. In the recent funding period funds supported

and has served as the departmental Executive Vice Chair

key lab personnel working on the project outlined;

since September 2015. In this role, he serves as an advisor to

fostered trainee education; and purchased necessary

the department chair, vice chairs, and senior administrative

supplies. This support remains vital to their efforts.

personnel on a broad array of matters of importance to the department including faculty recruitment and mentoring.

Dr. McCray is a practicing pediatric physician-scientist who staffs the Pediatric Pulmonary service, sees patients in clinic,

Dr. McCray’s most extensive and outstanding teaching

and takes night call. From 2010-2016 he served as interim

comes in the laboratory and his teaching of graduate and

Figure 2. Molecular correction of 3849+10 C>T in conditionally reprogrammed human CF airway epithelia. PCR evidence: top panel indicates the PCR primer sites. In middle panel “Wild” indicates non-CF cells, “3849” denotes control untreated reprogrammed cells as noted in text. “AAV” denotes mutant cells treated with the AAV2.5T vector encoding sgRNAs and the Sa Cas9 gene editing tools. The lower panel shows bioanalyzer results quantifying the abundance of the PCR products. Arrows indicate where the aberrant mutation product is expected to elute. Following the vector treatment, the aberrant transcript was no longer detected.

Figure 3. Primary human CF airway epithelial cells, F508del/F508del, were treated with dual AAV2.5T vectors delivering gene editing reagents and a HR template. Results shown include Surveyor (Cel I) nuclease assay (left side of upper left panel), RFLP (upper left, right side, white arrow denotes indel), and direct sequencing for insertion of CTT and a silent nucleotide modification that introduces a BsiW1 restriction enzyme site (lower panel). The bar graph summarizes the data.

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postdoctoral students. His curriculum vitae lists 12 undergraduate

His scientific career has been devoted to explaining the

students, 34 predoctoral students, 43 graduate students,

biological basis of cystic fibrosis and developing innovative

and 22 postdoctoral trainees whom he has supervised

ways to prevent or treat the associated lung disease.

or mentored. Many of the students and trainees he has mentored are now faculty members at the University of

His work in this area has led also to important discoveries

Iowa or other institutions. At the national level, he reviews

about host-pathogen interactions in the lung and about

grant applications for the NIH, Cystic Fibrosis Foundation,

the general topic of gene therapy for human diseases.

March of Dimes, and the national research organizations of England, Ireland, and the Netherlands. He reviews manuscripts for many of the best journals in biology and

MANUSCRIPTS

medicine. He serves on the Research and Research Training Committee of the Cystic Fibrosis Foundation, and he has served in the past on several committees for the American Society of Gene Therapy. Dr. McCray was awarded the Regents Award for Faculty Excellence in 2016.

1. Shah VS, Meyerholz DK, Tang XX, Reznikov L, Abou Alaiwa M, Ernst SE, Karp PH, Wohlford-Lenane CL, Heilmann KP, Leidinger MR, Allen PD, Zabner J, McCray PB Jr, Ostedgaard LS, Stoltz DA, Randak CO, Welsh MJ. Airway acidification initiates

Dr. McCray’s most influential contributions have been

host defense abnormalities in cystic fibrosis mice. Science. 2016 Jan

as a scientist. He is indisputably one of our country’s

29;351(6272):503-7. doi: 10.1126/science.aad5589. PMID: 26823428

leading investigators in the area of the pathogenesis and therapy of cystic fibrosis in infants and children.

2. Bartlett JA, Ramachandran S, Wohlford-Lenane CL, Barker CK, Pezzulo AA, Zabner J, Welsh MJ, Meyerholz DK, Stoltz DA, McCray PB Jr. Newborn Cystic Fibrosis Pigs Have a Blunted Early Response to an Inflammatory Stimulus. Am J Respir Crit Care Med. 2016 Mar 30. [Epub ahead of print] PMID: 27027566

3. Hornick AL, Li N, Oakland M, McCray PB Jr, Sinn PL. Human, Pig, and Mouse Interferon-Induced Transmembrane Proteins Partially Restrict Pseudotyped Lentiviral Vectors. Hum Gene Ther. 2016 May;27(5):354-62. doi: 10.1089/hum.2015.156. PMID: 27004832

4. Cooney AL, Abou Alaiwa MH, Shah VS, Bouzek DC, Stroik MR, Powers LS, Gansemer ND, Meyerholz DK, Welsh MJ, Stoltz DA, Sinn PL, McCray PB Jr. Lentiviral-mediated phenotypic correction of cystic fibrosis pigs. JCI Insight, 2016;1(14):e88730. doi:10.1172/jci.insight.88730.

5. Park JE , Kun Li K, Barlan A, Fehr AR, Perlman S, McCray PB Jr., Gallagher T. Proteolytic Processing of Middle East Respiratory Syndrome Coronavirus Spikes Expands Virus Tropism. PNAS, 2016 Oct 25;113(43):12262-12267. PMID: 27791014

6. Ramachandran S, Osterhaus SR, Parekh KR, Jacobi AM, Behlke MA, McCray PB Jr. SYVN1, NEDD8, and FBXO2 Regulate ΔF508-CFTR Ubiquitin-Mediated Proteasomal Degradation. J Biol Chem. 2016 Oct 18. pii: jbc.M116.754283. [Epub ahead of print] PMID: 27756846

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January 1, 2016 — December 31, 2016 FINANCIAL SUMMARY PAUL B. MCCRAY, JR., MD Balance Forward Revenue Disbursements to UI Total Revenue

$123,492 Travel 4.5%)

$85,973 Equipment (3%) Equipment Maintenance (13.3%)

Expenses Personnel $8,044 Lap Supplies/Animals $31,238 Travel 2,600 Equipment $1,718 Equipment Maintenance $7,683 Other $6,594 Total Expenses $57,877 Ending Balance

Other(11.4%) Personnel (13.9%) Lab Supplies Animals (54%)

$151,588

7. Li K, Wohlford-Lenane CL, Channappanavar R, Park J-E, Bair TB,

4. Krishnamurthy S, Locke T, Li X, Yan WX, Ran FA, Zhang F. Zabner J,

Flaherty HA, Gallagher T, Meyerholz DK, Perlman S, McCray PB Jr.

McCray P. Characterization of CRISPR/Cas9 Editing of Two Different

Mouse Adapted MERSCoronavirus Causes Lethal Lung Disease in Human

Classes of Cystic Fibrosis Mutations and their Repair by Two Distinct

DPP4 Knock-in Mice. PNAS, in revision.

Repair Mechanisms. Pediatr Pulmonol. Suppl 45: 198 [Abstract #12], 2016.

ABSTRACTS

5. Strub MD, Hodos RA, Meleshkevitch EA, Boudko DY, Bridges RJ, Dudley JT, McCray P. Integrative Genomic SignatureBased Approach to Discover Drugs for DF508-CFTR Rescue.

1. Li K, Wohlford-Lenane CL, McCray PB. The Middle East Respiratory

Pediatr Pulmonol. Suppl 45:216 [Abstract #62], 2016.

Syndrome Coronavirus (MERS-CoV) Exhibits Bilateral Entry and Exclusively Apical Egress in Well-Differentiated Human Airway Epithelial Cells. Am J Resp Crit Care Med.193: A4457 [Abstract #907], 2016.

6. Bartlett JA, Berkebile A, Rogan MP, Weldon S, Abou Alaiwa MH, Welsh M, Taggart C, McCray P. Impaired Antiviral Activity and Increased Cathepsin Activity in Airway Secretions from Newborn

2. Wohlford-Lenane CL, Li K, Meyerholz DK, Perlman S,

CF Pigs. Pediatr Pulmonol. Suppl 45: 237 [Abstract #120], 2016.

McCray PB. Mouse Models to Study Middle East Respiratory Syndrome Disease Pathogenesis and Treatment. Am J Resp Crit Care Med.193:A4458 [Abstract #908], 2016.

3. Sinn P, Cooney A, Abou Alaiwa, MH, Thornell I, Zabner J, Meyerholz DK, Welsh M, Stoltz DA, McCray P. Integrating Viral Vectors for Cystic Fibrosis Gene Therapy in CF Pigs. Pediatr Pulmonol. Suppl 45: 266 [Abstract #196], 2016.

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JANE S. PAULSEN, PHD Dr. Jane Paulsen’s research focuses on the

ROY J. CARVER CHAIR IN NEUROSCIENCE

translation of findings in brain research to

DIRECTOR, UNIVERSITY OF IOWA HUNTINGTON

Huntington’s disease.

application for persons living with brain diseases, such as Alzheimer’s, Parkinson’s or

DISEASE CENTER OF EXCELLENCE DIRECTOR, DIVISION OF NEUROPSYCHOLOGY PROFESSOR, DEPARTMENT OF PSYCHIATRY

Funds from the Roy J. Carver Charitable Trust were

PROFESSOR, DEPARTMENT OF NEUROLOGY

used in 2016 to cover the salary cap for Dr. Paulsen. To clarify, Dr. Paulsen had several active research grants in 2016. As a consequence her time is tightly committed to research progress funded through her grants. Support from the Carver Trust allows Dr. Paulsen to facilitate additional research networks and infrastructure. Dr. Paulsen utilized her Carver Trust Support in 2016 to embark upon a new area of research, CADASIL. Cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL) is the most common heritable cause of stroke and vascular dementia in adults. Vascular dementia is the second most common type of cognitive disorders in mid and late life (following Alzheimer’s disease). Learning how to prevent or slow cognitive losses would benefit from advanced understanding of the causes of this dementia. Study of persons at 100 percent risk of a known vascular dementia could help us learn how the disease starts and how it progresses. Knowledge of these aspects of the disease could lead to treatments to delay onset of the cognitive decline or prevent the disease from leading to dementia. Dr. Paulsen’s new interest in CADASIL has the unique potential to provide knowledge for the fields of stroke and vascular dementia. Although this new research area is just beginning, Dr. Paulsen has already recruited colleagues from the intramural research program of the National Institute of Heart Lung and Blood to work with her.

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In 2016, the Paulsen lab had ten collaborative research grants (Scherzer, Gusella, Wang, Chelsky, Turner, Potkin, Carlozzi, Miller, Westin, Max), four Iowa only HD research grants (Enroll, COE, Proteomics, Exercise) and four clinical trials of experimental therapeutics (called HART, ARC, SIGNAL, and LEGATO), for a total of 18 active grants. Additionally, Dr. Paulsen mentored four graduate students/fellows (Misuri, Esperanza, Ciarochi, Caprehan) and ten junior faculty members (Downing, Turner, Rao, Ipek, Sui, Kim, Westerfeld, Lourens, Schaffer, Schultz). Dr. Paulsen was invited as a keynote speaker by two Midwest Colleges of Medicine in 2016. She presented her neuroscience research in Huntington’s disease at the University of Nebraska in Omaha and the Univeristy of South Dakota in Vermillion and Sioux Falls, South Dakota. While on these travels to present to the professors, fellows, and medical students at these schools, she also made time to meet with HD families and present workshops on addressing difficult behaviors in HD Dementia. Dr. Paulsen was invited to be the keynote speaker for the Tawain Movement Disorder Society and was invited to teach at the National Taiwan University for four days. She was also invited by the European HD Network to present her work at Den Hague in the Netherlands where she educated colleagues on the psychiatric aspects of HD. Dr. Paulsen was invited to become a member of the Clinical Neuroscience and Neurodegeneration Study Section of the Brain Disorders and Clinical Neuroscience (BDCN) Center for Scientific Review at the National Institutes of Health. Additionally she was invited to participate as a speaker and panel member for 2016 Alzheimer’s Disease and Related Disorders meeting which emphasized Lewy Body diseases and Frontotemporal Dementias.

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6. Harrington, D. L., et al. (2016). “Cross-sectional and longitudinal multimodal structural imaging in prodromal Huntington’s disease.” Mov Disord 31(11): 1664-1675.

7. Forbes, J. L., et al. (2016). “An Open-Source Label Atlas Correction Tool and Preliminary Results on Huntingtons Disease Whole-Brain MRI Atlases.” Front Neuroinform 10: 29.

8. Huntington Study, G., et al. (2016). “Effect of Deutetrabenazine on Chorea Among Patients With Huntington Disease: A Randomized Clinical Trial.” JAMA 316(1): 40-50.

9. Holtbernd, F., et al. (2016). “Longitudinal Changes in the Motor Learning-Related Brain Activation Response in Presymptomatic Huntington’s Disease.” PLoS One 11(5): e0154742.

10. Faria, A. V., et al. (2016). “Linking white matter and deep gray matter alterations in premanifest Huntington

PUBLICATIONS

1. Muralidharan, P., et al. (2016). “Bayesian Covariate Selection in Mixed-Effects Models For Longitudinal Shape Analysis.” Proc IEEE Int Symp Biomed Imaging 2016: 656-659.

2. Shaffer, J. J., et al. (2017). “Longitudinal diffusion changes in prodromal and early HD: Evidence of white-matter tract deterioration.” Hum Brain Mapp.

3. Downing, N. R., et al. (2016). “Phenotype Characterization of HD Intermediate Alleles in PREDICT-HD.” J Huntingtons Dis 5(4): 357-368.

4. Long, J. D., et al. (2016). “Validation of a prognostic index for Huntington’s disease.” Mov Disord.

5. Ciarochi, J. A., et al. (2016). “Patterns of Co-Occurring Gray Matter Concentration Loss across the Huntington

disease.” Neuroimage Clin 11: 450-460

11. Dixon, B. S., et al. (2016). “Cognitive changes associated with switching to frequent nocturnal hemodialysis or renal transplantation.” BMC Nephrol 17: 12.

12. Carvalho, J. O., et al. (2016). “The impact of oculomotor functioning on neuropsychological performance in Huntington disease.” J Clin Exp Neuropsychol 38(2): 217-226.

13. Epping, E. A., et al. (2016). “Longitudinal Psychiatric Symptoms in Prodromal Huntington’s Disease: A Decade of Data.” Am J Psychiatry 173(2): 184-192.

14. Beglinger, L. J., et al. (2015). “Practice Effects and Stability of Neuropsychological and UHDRS Tests Over Short Retest Intervals in Huntington Disease.” J Huntingtons Dis 4(3): 251-260.

Disease Prodrome.” Front Neurol 7: 147.

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

JANE S. PAULSEN, PHD Balance Forward

$277,158

Revenue Disbursements to UI Total Revenue

$93,810

Other (0.1%)

Expenses Personnel $5,260 Other $7 Total Expenses $5,267 Ending Balance

Personnel (99.9%)

$365,701

15. Liu, D., et al. (2015). “Motor onset and diagnosis in Huntington disease

21. Kim, J. I., et al. (2015). “Multivariate clustering of progression

using the diagnostic confidence level.” J Neurol 262(12): 2691-2698.

profiles reveals different depression patterns in prodromal Huntington disease.” Neuropsychology 29(6): 949-960.

16. Long, J. D., et al. (2015). “Multivariate prediction of motor diagnosis in Huntington’s disease: 12 years of PREDICT-HD.” Mov Disord 30(12): 1664-1672.

22. Wassef, S. N., et al. (2015). “T1rho imaging in premanifest Huntington disease reveals changes associated with disease progression.” Mov Disord 30(8): 1107-1114.

17. Musso, M., et al. (2015). “Intra-individual Variability in Prodromal Huntington Disease and Its Relationship to Genetic Burden.” J Int Neuropsychol Soc 21(1): 8-21.

23. Kim, J. I., et al. (2015). “Performance of the 12-item WHODAS 2.0 in prodromal Huntington disease.” Eur J Hum Genet 23(11): 1584-1587

18. Kim, R. E., et al. (2015). “Preliminary analysis using multi-atlas labeling

24. Carlozzi, N. E., et al. (2015). “Validity of the 12-item World

algorithms for tracing longitudinal change.” Front Neurosci 9: 242.

Health Organization Disability Assessment Schedule 2.0

19. Matsui, J. T., et al. (2015). “Prefrontal cortex white matter tracts in prodromal Huntington disease.” Hum Brain Mapp 36(10): 3717-3732.

20. Harrington, D. L., et al. (2015). “Network topology and

25. Williams, J. K., et al. (2015). “Everyday cognition in prodromal Huntington disease.” Neuropsychology 29(2): 255-267.

26. Panegyres, P. K., et al. (2015). “Factors influencing the clinical

functional connectivity disturbances precede the onset of

expression of intermediate CAG repeat length mutations of

Huntington’s disease.” Brain 138(Pt 8): 2332-2346.

the Huntington’s disease gene.” J Neurol 262(2): 277-284.

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GEORGE B. RICHERSON, MD, PHD The overall goal of Dr. George Richerson’s

ROY J. CARVER CHAIR IN NEUROSCIENCE CHAIR AND DEPARTMENT EXECUTIVE OFFICER,

research is to study how the brain controls breathing and how abnormal breathing can contribute to various diseases. For example,

DEPARTMENT OF NEUROLOGY DIRECTOR, CLINICAL NEUROSCIENTIST TRAINING PROGRAM IN NEUROLOGY

sudden unexpected death in epilepsy (SUDEP) and sudden infant death syndrome (SIDS) are both linked to defects in neurons

PROFESSOR, DEPARTMENT OF NEUROLOGY

that supply the neurotransmitter serotonin

PROFESSOR, DEPARTMENT OF MOLECULAR

(5-HT) to the brain. He has shown that 5-HT

PHYSIOLOGY AND BIOPHYSICS

neurons are essential to control breathing to maintain normal levels of carbon dioxide (CO2) and pH.

Funds from the Roy J. Carver Charitable Trust supporting the Roy J. Carver Chair in Neuroscience were used to provide salary and fringe support to Dr. George Richerson.This support allowed Dr. Richerson protected time to continue his research throughout the year. SUDEP and SIDS are major public health problems. Patients with epilepsy have a risk of sudden unexpected death that is more than 20 times greater than the general population. In the state of Iowa, it is estimated that there are 45,000 people with epilepsy, and 15,000 of them are refractory to treatment (i.e. they continue to have frequent seizures even though they are taking two or more anticonvulsant drugs). Refractory epilepsy patients have the highest risk of SUDEP, with as many as 50 percent dying from this cause. Many of these deaths occur in otherwise healthy young people. SIDS is also very common. Despite the prevalence having decreased in half by the “Back to Sleep” campaign, it is still the most common cause of postneonatal infant death in industrialized countries, occurring in 0.6 per 1000 live births. It too occurs in otherwise seemingly healthy individuals.

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In the last year, Dr. Richerson has made significant

Experiments have been performed studying the effect of diet

progress in his scientific goals. He continues to work on

on seizures and SUDEP in mice. It is known that some patients

a multicenter NIH/NINDS grant to study SUDEP with

can benefit from a diet that is high in fat, low in carbohydrates

investigators at 14 institutions across the world. This

and adequate in protein. This is called a ketogenic diet because

award is being used to perform research in rodents and in

it generates ketone bodies (acetone, beta-hydroxybutyrate and

humans to understand the causes of SUDEP, to identify

acetoacetate). However, this diet does not work for everyone

biomarkers of high risk patients, and to find ways to

and it is very difficult to stay compliant with the diet as it is not

prevent death. The work is being done in collaboration

palatable for many patients. Preliminary data indicate that a

with members of the Departments of Neurology, Internal

diet with a limited subset of components of a ketogenic diet

Medicine (Pulmonary), Psychiatry and Neurosurgery at the

can be as effective in preventing seizures and SUDEP in mice,

University of Iowa, and with neurologists at Northwestern

even when it does not cause ketosis. The components of this

University and University of Southern California.

diet that are most effective are being defined so that a dietary supplement might be formulated for use as an anticonvulsant.

Part of this work focuses on Dravet Syndrome, which is a devastating, untreatable infantile-onset form of epilepsy with

Patients with epilepsy are routinely admitted to Epilepsy

a high rate of SUDEP. Dr. Richerson’s group has determined

Monitoring Units at UI Hospitals and Clinics and

that children with Dravet Syndrome have severe breathing

Northwestern University. Recordings are made of their

abnormalities after seizures, and these are likely to be the cause

EEG, breathing and EKG during seizures. Information is

of death in some cases of SUDEP. Experiments in a mouse

gathered about how their breathing is affected by seizures

model of Dravet Syndrome have shown that seizures cause

and by changes in CO2. Sadly, two of the patients studied

death due to respiratory arrest, and have led to identification

at UI Hospitals and Clinics and Northwestern University

of a dietary treatment that prevents seizures and death in these

in the last 1.5 years have died of SUDEP while at home

mice. This diet may benefit children with Dravet Syndrome.

in bed. Although this is incredibly unfortunate, the data obtained from these patients provides important insight into

In a paper published in Neuroscience, it was shown that 5-HT

why these patients died, and appear to point at a biomarker

neurons do not respond to pH in newborn mice, and mice do

that can be used in the future to identify those epilepsy

not increase their breathing in response to CO2 until they are

patients who are at highest risk of dying from SUDEP.

2-3 weeks old. After that age mice become very sensitive to a rise in CO2. These findings help to interpret experiments on

Dr. Richerson is now widely recognized as a leader in the

newborn rodents, and are relevant to the mechanisms of SIDS.

international epilepsy community and the SUDEP field. Dr. Richerson was Vice-Director of the PAME 2016 meeting in

In a paper published in The Journal of Neuroscience Dr.

Alexandria, VA, and will be the Director of the PAME 2018

Richerson, along with collaborators from Yale University

meeting. These positions reflect the respect of the epilepsy

and the University of Iowa, reported that seizures induced

community for the research performed in his laboratory.

in the hippocampus of rats led to respiratory arrest. This

He is a permanent member of the NIH, NINDS, NST-1

coincided with inhibition of 5-HT neurons in the medulla.

study section responsible for reviewing career development

That observation is consistent with the hypothesis that

(K08 & K23) awards for junior physician-scientists.

seizures cause apnea by inhibiting the respiratory network by suppressing the excitatory drive to breath by 5-HT neurons. Current work is aimed at determining whether seizures inhibit 5-HT neurons by causing activation of descending projections from the amygdala. If so, that would explain observations reported in 2015 from human epilepsy patients showing that activation of the amygdala causes inhibition of breathing.

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GRANTS, PUBLICATIONS, SYMPOSIA

Grants 1. NIH/NINDS – U01 NS090414. 10/1/2014 to 9/30/2019. Title: SUDEP - Respiratory and arousal mechanisms. Direct costs to U. of Iowa: $430,000 per year; $2,150,000 total.

2. NIH/NINDS – R25 NS079173 Residency Training Grant. 4/1/2012 – 6/30/2017. Roy J. & Lucille A. Carver College of Medicine Clinical Neuroscientist Training Program (CNS-TP). Direct costs: $70,000 per year.

3. NIH/NINDS – U01 NS090407. 10/1/2014 – 7/31/2019. Title: Center for SUDEP Research / SUDEP Center Without Walls. Subcontract for Clinical Research on SUDEP. Direct costs: $34,428 per year.

4. NIH/NINDS - K08 NS069667. 7/1/11-6/30/16. Mentor for Gordon Buchanan, MD, PhD

5. NIH/NINDS - K08. 7/1/12-6/30/17. Mentor for Nandakumar Narayanan, MD, PhD

6. NIH/NINDS – K12. 7/1/16-6/30/18. Mentor for Brian Dlouhy, MD.

Original Reports 1. Cerpa, V.J., Y. Wu, E.U. Bravo, F.A. Teran, R.S. Flynn & G.B. Richerson. Medullary 5-HT neurons: Switch from respiratory drive to chemoreception during postnatal development. Neurosci (in press), 2016. [PMID: 27619736]

2. Zhan, Q., G. Buchanan, J. Motelow, J. Andrews, P. Vitkovskiy, W. Chen, F. Serout, A. Gummadavelli, A. Kundishora, M. Furman, W. Li, X. Bo, G.B. Richerson, and H. Blumenfeld. Impaired serotonergic brainstem function during and following seizures. J Neurosci, 36(9):2711-22, 2016. [PMID: 26937010]

Reviews 1. Buchanan, G.F. & G.B. Richerson. Epilepsy: A dietary supplement for SUDEP prevention? Nature Rev Neurol 12(9):495-6, 2016. [PMID: 27514288]

2. Devinsky, O., D.C. Hesdorffer, D. Thurman, S. Lhatoo, G.B. Richerson. Sudden Unexpected Death in Epilepsy: Epidemiology, Mechanisms & Prevention. Lancet Neurology, 15(10):1075-88, 2016. [PMID: 27571159]

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

GEORGE B. RICHERSON, MD, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$64,111

$96,624 Personnel (100%)

Expenses Personnel $125,484 Total Expenses $125,484 Ending Balance

$35,251

3. Richerson, G.B., D. Boison, C. Faingold & P. Ryvlin. SUDEP: From unwitnessed fatality to witnessed rescue: pharmacological

6. Neurology Grand Rounds. “SUDEP: Neither sudden nor unexpected?” Department of Neurology, University of Iowa, Iowa City, IA, 5/2016.

intervention. Epilepsia, 57(Suppl 1):35-45, 2016. [PMID: 26749015]

Symposia and Invited Talks 1. Seminar Speaker. “SUDEP and SIDS: Is there a final common

7. Invited Speaker. “Developing a research career.” In: “Navigating your Career: All Aboard!” American Academy of Neurology Meeting, Vancouver, BC, Canada. 4/2016.

pathway for sudden death?” Center for Integrated Brain Research, Seattle Children’s Hospital. Seattle, WA, 9/2016.

8. Epilepsy Lecture Series. “SUDEP: Neither sudden nor unexpected?” Comprehensive Epilepsy Center,

2. Neurology Grand Rounds. “SUDEP: Neither sudden nor unexpected?”

Northwestern University, Chicago, IL, 4/2016.

Department of Neurology, University of Washington, Seattle, WA, 9/2016. 9. Invited Speaker. “Nuts and Bolts of Building a Dual Career 3. Symposium Speaker. “Changes in respiratory control induced by seizures.” In: “What are the events that occur during and after a

as a Clinician-Researcher.” NINDS R25 Physician Scientist Training Program Meeting, Washington, DC, 3/2016.

seizure that cause death in SUDEP?” Partners Against Mortality in Epilepsy (P.A.M.E.) 3rd Biannual meeting. Alexandria, VA 6/2016.

10. Symposium Speaker. “When Life Support Fails—Insights into the Mechanisms of Cardiorespiratory Collapse after Seizures.” In: “UUUgh!

4. Conference Organizer and Speaker. “Comprehensive epilepsy management at Iowa: A program for patients,

The Unexpected, Unexplained and often Undetermined.” National Association of Medical Examiner’s Interim meeting, Las Vegas, NV, 2/2016.

families and the community.” Coralville, IA, 5/2016. 11. Invited participant. “New Approaches to Enhance the Physician 5. Invited Speaker. “Epilepsy: Causes and consequences.” Mini

Scientist Workforce.” NIH workshop. Bethesda, MD, 2/2016.

Medical School. University of Iowa, Iowa City, IA, 5/2016.

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VAL C. SHEFFIELD, MD, PHD The overall goal of the research of Dr. Val

ROY J. CARVER CHAIR IN MOLECULAR GENETICS

Sheffield is to understand the genetic

DIRECTOR, DIVISION OF MEDICAL GENETICS

management and treatment. Dr. Sheffield’s

DIRECTOR, INTERDEPARTMENTAL RESEARCH

laboratory specifically studies Mendelian

PROGRAM IN HUMAN MOLECULAR GENETICS PROFESSOR, DEPARTMENT OF PEDIATRICS—

human genetic diseases and improve their

genetic disorders that provide insight into common human disorders including

MEDICAL GENETICS PROFESSOR, DEPARTMENT OF

basis and disease mechanisms of specific

OPHTHALMOLOGY AND VISUAL SCIENCES

obesity, high blood pressure (hypertension), diabetes, and blindness.

During the past year, some funds from the Roy J. Carver Charitable Trust Endowed Chair were used for salary support for Dr. Sheffield. This allows other funds to go to paying for his research experiments. In addition, endowed funds go to support the operating costs of his laboratory including laboratory supplies. This endowed chair has greatly lessened the impact of the ending of Howard Hughes Medical Institute funding (after eighteen years) and is allowing his research to continue at a high level with less interruption. Genetic Basis and Pathophysiology of Genetic Diseases To understand the genetic basis and the pathophysiology of specific human genetic diseases and improve their management, Dr. Sheffield’s goals include: 1. identify the molecular components of hereditary diseases; 2. determine the functions of the genes/ proteins associated with these disorders; 3. determine genetic and protein interactions, as well as define the protein complexes and networks that contribute to these diseases;

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4. develope animal models to aid in determining the phenotype-specific pathophysiology; and

trains young faculty at the University of Iowa and helps them develop research careers and obtain funding. He serves as director of the Division of Medical Genetics, which

5. utilize animal models to develop interventions and treatments.

provides patient care to both pediatric and adult patients who have genetic disorders. Dr. Sheffield also serves on various advisory committees, including as a member of the Advisory Council for the National Human Genome

In the past year, the laboratory has utilized a variety of

Research Institute (NHGRI); member of the panel of experts

strategies to develop treatments for human genetic diseases

for the Human Health and Heredity in Africa (H3Africa)

with a focus on using mouse models to explore in treatment

program, and advisor to the Intellectual Disabilities and

of human disorders. The treatment strategies include:

Development Research Center at Boston Children’s Hospital.

1. gene therapy; and 2. treatment utilizing inducible pluripotent stem cells (iPSCs). The comprehensive approach being taken is leading to the identification and validation of treatment paradigms for eye diseases, obesity, hypertension, and diabetes. Dr. Sheffield’s laboratory has worked on a number of genetic disorders in the past year in which his laboratory has contributed to gene identification and elucidation of disease mechanisms. This work has included inherited eye diseases, specifically retinopathies and glaucoma, as well as non-eye disorders. Dr. Sheffield’s laboratory has also made progress using mouse models of disease to develop a novel treatment for glaucoma, a common cause of human blindess, using an approach known as genome editing, which alters the actual DNA of cells involved in the disease. In addition, the laboratory continues to make progress in understanding hydrocephalus, a common human birth defect, and developing treatments for hydrocephalus. Dr. Sheffield’s accomplishments outside the research arena include: Mentoring undergraduate and graduate students, the latter in helping them obtain their PhD degrees. During the past year, four of his PhD students passed their comprehensive examinations, which allows them to become PhD candidates. He also provides research opportunities for post-doctoral fellows and medical students within his laboratory. His mentoring of one medical student propelled this student toward obtaining both PhD and MD degrees, and being accepted to the joint MD/PhD program. He

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PUBLICATIONS

1. Lu W, Hu H, Sevigny J, Gabelt BT, Kaufman PL, Johnson EC, Morrison JC, Zode GS, Sheffield VC, Zhang X, Laties AM, Mitchell CH. Rat, mouse, and primate models of chronic glaucoma show sustained elevation of extracellular ATP and altered purinergic signaling in the posterior eye. Invest Ophthalmol Vis Sci 2015 1;56(5):3075-83. PMID 260024091

2. Zhu W, Gramlich OW, Laboissonniere L, Jain A, Sheffield VC, Trimarchi JM, Tucker BA, Kuehn MH. Transplantation of iPSCderived TM cells rescues glaucoma phenotypes in vivo. Proc Natl Adac Sci U S A 2016 21;113(25):E3492-500 PMID 27274060

3. Starks RD, Beyer AM, Guo DF, Boland L, Zhang Q , Sheffield VC, Rahmouni K. Regulation of insulin receptor trafficking by Bardet-Biedl Syndrome proteins. PLoS Genet 2015 23;11(6):e1005311 doi:10.1371 PMID: 26103456

4. Datta P, Allamargot C, Hudson JS, Andersen EK, Bhattarai S, Drack AV, Sheffield VC, Seo S. Accumulation of non-outer segment proteins in the outer segment underlies photoreceptor degeneration in Bardet-Biedl syndrome. Proc Natl Acad Sci U S A. 2015 Jul 27 pil: 201510111 PMID 26216965

5. Haziza S, Magnani R, Lan D, Keinan O, Saada A, Hershkovitz E, Yanay N, Cohen Y, Nevo Y, Houtz RL, Sheffield VC, Golan H, Parvari R. Calmodulin methyltransferase is required for growth, muscle strength, somottosensory development and brain function. PLoS Genet 2015 6;11(8):e1005388 PMID 26247364

6. Muhammad E, Levitas A, Singh SR, Braiman A, Ofir R, Etzion S, Sheffield VC, Etzion Y, Carrier L, Parvari R. PLEXHM2 mutation leads to abnormal localization of lysosomes, impaired autophagy flux and associates with recessive dilated cardiomyopathy and left ventricular noncompaction. Hum Mol Genet 2015 [Epub ahead of print] PMID: 26464484

7. Guo DF, Cui H, Zhang Q , Morgan DA, Thedens DR, Nishimura D, Grobe JL, Sheffield VC, Rahmouni K. The BBSome controls energy homeostasis by mediating the transport of the leptin receptor to the plasma membrane. PLoS Genet, 2016 12(2):e1005890 PMID 26926121

8. Heon E, Kim G, Qin S, Garrison JE, Tavares E, Vincent A, Nuangchamnong N, Scott CA, Slusarski DC, Sheffield VC. Mutations

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

VAL C. SHEFFIELD, MD, PHD Balance Forward

$231,002

Revenue Disbursements to UI Total Revenue

$156,681

Expenses Lab Supplies/Animals Total Expenses Ending Balance

Lab Supplies/ Animals (100%)

$145,025 $145,025 $242,658

in C80RF37 cause Bardet Biedl syndrome (BBS21). Hum Mol Genet 2016, pii:ddw096 (Epub ahead of print) PMID: 27008867

9. Preist JR, Osoegawa K, Mohammed N, Nanda V, Kundu R, Schultz K, Lammer EJ, Girirajan S, Scheetz T, Waggott D, Haddad F, Reddy S, Bernstein D, Burns T, Steimle JD, Yang XH, Moskowitz IP, Hurles M, Lifton RP, Nickerson D, Barnshad M, Eichier EE, Mital S, Sheffield VC, Quertemous T, Gelb BD, Portman M, Ashley EA. De Novo and rare variants at multiple loci support the oligogenic origins of atrioventricular septal heart defects. PLos Genet 2016 12(40:e2005963 doi: 10.1371/journal.pgen eCollection PMIC 27058611

10. Reish O, Aspit L, Zouella A, Roth Y, Polak-Charcon S, Baboushkin T, Benyamini L, Scheetz TE, Mussaffi H, Sheffield VC, Parvari R. A homozygous Nme7 mutation is associated with situe inversus totalis. Hum Mat 2016 37(8):727-31 PMID 27060491

11. Kawasaki M, Izu Y, Hayata T, Ideno H, Niifuji A, Sheffield VC, Ezura Y, Noda M. Bardet-Biedl syndrome 3 regulates development of cranial base midline structures. Bone 2016 pii:S8756-3282(16)30043-6 PMID: 27170093

12. Heon E, Kim G, Qin S, Garrison JE, Tavares E, Vincent A, Nuangchamnong N, Scott CA, Slusarski DC, Sheffield VC. Mutations in C80RF37 cause Bardet Biedl syndrome (BBS21). Hum Mol Genet. 2016 1;25(11):2283-2294 PMID:27008867

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CURT D. SIGMUND, PHD Dr. Curt Sigmund’s research uses state-

ROY J. CARVER CHAIR IN HYPERTENSION

of-the-art molecular genetic techniques

CHAIR AND DEPARTMENT EXECUTIVE OFFICER,

physiological pathways regulating blood

DEPARTMENT OF PHARMACOLOGY PROFESSOR, DEPARTMENT OF PHARMACOLOGY PROFESSOR, DEPARTMENT OF MOLECULAR

to alter the mouse genome to uncover the molecular, genetic, cellular, and

pressure and when they go awry— hypertension.

PHYSIOLOGY & BIOPHYSICS PROFESSOR, DEPARTMENT OF INTERNAL MEDICINCE

Dr. Sigmund was named the Roy J. Carver Chair in Hypertension Research in July 2008. In previous years, funding has been used entirely to defray salary support for Dr. Sigmund. This has freed up substantial funding from NIH grants to be used toward the funded research projects, and provided protected time for Dr. Sigmund to devote to his research program. In 2016, funds from the Roy J. Carver Charitable Trust supporting the Roy J. Carver Chair in Hypertension Research were used to 1) defray partial salary for Dr. Sigmund; 2) partially fund the salaries, fringe benefits, and tuition of a graduate student and postdoctoral fellow in Dr. Sigmund’s laboratory; 3) purchase and support the per diem expenses of experimental mice; 4) repair and purchase of new equipment; and 5) the largest amount, to purchase research supplies and reagents to conduct experiments in the laboratory. This has been a particularly productive year both in terms of research results leading to publications and for acquisition of new research grants. First, the NIH grant application entitled “Role of PPARg and the PPARg Target Gene RBP7 in the Endothelium” was funded (on its first submission) on April 1, 2016, for four years at a total budget of $2,512,664. Funds from both the Carver Chair and RPOE were essential for the acquisition of preliminary data supporting this grant. With this NIH R01 grant funded, Dr. Sigmund’s portfolio of funding now includes the following: • NIH NHLBI R01 “Hypertension: Role of Smooth Muscle Cullin-3 and the CRL3 Complex” (funded through March 2019)

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• NIH NHLBI R01 “Role of PPARg and the

Dr. Sigmund’s research and scholarship as the part of

PPARg Target Gene RBP7 in the Endothelium”

the Carver RPOE and the Carver Chair in Hypertension

(funded through March 2020)

Research has resulted in several lectures and seminars in the past year at universities and at national and international

• NIH NHLBI P01 (PPG) “Genetic and Signaling Mechanisms in the Central Regulation of Blood Pressure” (funded through March 2018) • American Heart Association Strategically Focused Hypertension Research Network (funded through March 2019) In aggregate this represents over $15 million in research support to the Sigmund laboratory and UIHC Center for Hypertension Research. Importantly, Dr. Sigmund’s Program Project Grant entitled “Genetic and Signaling Mechanisms in the Central Regulation

scientific meetings. Dr. Sigmund was chair of the prestigious Gordon Research Conference on Angiotensin held February 21-26, 2016 in Il Ciocco, Barga, Italy. He was selected as the Distinguished Lecturer in Physiological Genomics by the American Physiological Society at its annual meeting, and gave an invited lecture on Recent Advances in Hypertension at the annual meeting of the AHA Council on Hypertension. Dr. Sigmund presented an invited seminar at University of Alabama-Birmingham in 2016 and is scheduled to present at the University of Virginia, Weill Cornell Medical College, Medical College of Wisconsin, and Vanderbilt University in early 2017. Dr. Sigmund will also continue his four-year term as Chair of the Publications Committee of the American Physiological Society.

of Blood Pressure” will enter is final year of the current

Studies in the Sigmund laboratory have resulted in

funding cycle on April 1, 2017. The PPG team is working very

15 independent and collaborative publications in

hard to develop a competitive PPG renewal application that is

2016.

expected to be submitted in September 2017. Support from the Carver Trust (both this Chair and the RPOE) was instrumental in the funding of this program five years ago, and will be instrumental in its competitive renewal later this year. Indeed, with support from the Carver Trust, we published several seminal research papers defining novel mechanisms controlling blood pressure, hypertension, and obesity-related hypertension. The Sigmund laboratory published 15 research papers and reviews in 2016. Several of these papers provide evidence of productivity on Dr. Sigmund’s NIH Program Project Grant. For example, in the paper entitled “Selective Deletion of the Brain-Specific Isoform of Renin Causes Neurogenic Hypertension,” we defined a novel pathway that activates the renin-angiotensin system in the brain leading to hypertension. It defines a molecular pathway for the cause of hypertension in some types of patients with difficult to treat (also called refractory or resistant) hypertension. Similarly, in a collaboration between the Sigmund and Grobe laboratories (funded through the PPG and Carver Trust), we published a paper in the high impact journal Cell Reports entitled “Suppression of Resting Metabolism by the Angiotensin AT2 Receptor,” This will likely be considered a seminal paper as it defines for the first time a cross talk between renin-angiotensin action in the brain with its action in adipose (fat) tissue. This study has important implications for hypertension in obesity.

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PUBLICATIONS

1. Ketsawatsomkron, P., Keen, H.L., Davis, D.R., Lu, K-T., Stump, M., De Silva, T.M., Hilzendeger, A.M., Grobe, J.L., Faraci, F.M., Sigmund, C.D. A Protective Role for TIMP-4, a Novel PPARg Target Gene, in Smooth Muscle in DOCA-Salt Hypertension. Hypertension 67: 214-222, 2016.

2. Mukohda. M., Stump, M., Ketsawatsomkron, P., Hu, C., Quelle, F.W., Sigmund, C.D. Endothelial PPARg Provides Vascular Protection from IL-1β-Induced Oxidative Stress. Am J Physiol Heart Circ Physiol. 310: H39-H48, 2016.

3. Hu, C., Lu, K-T., Mukohda, M., Davis, D.R., Faraci, F.M., Sigmund, C.D. Interference with PPARg in Endothelium Accelerates Angiotensin II-induced Endothelial Dysfunction. Physiological Genomics 48: 124-134, 2016.

4. Chu, Y., Lund, D.D., Doshi, H., Keen, H.L., Knudtson, K.L., Funk, N.D., Shao, J.Q., Cheng, J., Hajj, G.P., Zimmerman, K.A., Davis, M.K., Brooks, R.M., Chapleau, M.W., Sigmund, C.D., Weiss, R.M., Heistad, D.D. Fibrotic Aortic Valve Stenosis in Hypercholesterolemic/ Hypertensive Mice. Arterioscler Thromb Vasc Biol. 36:466-474, 2016.

5. Lu, K-T., Keen, H.L., Weatherford, E.T., Sequeira-Lopez, M.L.S., Gomez, R.A., Sigmund, C.D. Estrogen Receptor α is Required for Maintaining Baseline Renin Expression. Hypertension 67: 992-999, 2016. (PMC4833511)

6. Stump, M., Guo, D-F., Lu, K-T., Mukohda, M., Liu, X., Rahmouni, K. Sigmund, C.D. Effect of Selective Expression of Dominant Negative PPARg in Proopiomelanocortin Neurons on the Control of Energy Balance. Physiological Genomics 48: 491-501, 2016.

7. Muta, K., Morgan, D.A., Grobe, J.L., Sigmund, C.D., Rahmouni, K. mTORC1 Signaling Contributes to Drinking but Not Blood Pressure Responses to Brain Angiotensin II. Endocrinology. 157: 3140-3148, 2016.

8. Muntner, P., Becker, R.C., Calhoun, D., Chen, D., Cowley, A.W. Jr, Flynn, J.T., Grobe, J.L., Kidambi, S., Kotchen, T.A., Lackland, D.T., Leslie, K.K., Li,Y., Liang, M., Lloyd, A., Mattson, D.L., Mendizabal, B., Mitsnefes, M., Nair, A., Pierce, G.L., Pollock, J.S., Safford, M.M., Santillan, M.K., Sigmund, C.D., Thomas, S.J., Urbina, E.M. Introduction to the American Heart Association’s Hypertension Strategically Focused Research Network. Hypertension. 67:674-680, 2016.

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

CURT D. SIGMUND, PHD Balance Forward

$55,457

Revenue Disbursements to UI Total Revenue

$95,613

Expenses Personnel $8,532 Lab Supplies/Animals $43,266 Equipment $1,355 Equipment Maintenance $257 Other $4,605 Total Expenses $58,016 Ending Balance

Other (7.9%)

Personnel (14.7%)

Equipment Maintenance (.4%)

Equipment (2.3%)

Lab Supplies/ Animals (74.6%)

$93,055

9. Song, K., Stuart, D., Abraham, N., Wang, F., Wang, S., Yang, T., Sigmund, C.D., Kohan, D.E., Ramkumar, N. Collecting

Grumbach, I.M. Role of CaMKII in Ang-II-dependent Small Artery Remodeling. Vascular Pharmacology 87: 172-179, 2016.

Duct Renin Does Not Mediate DOCA-Salt Hypertension or Renal Injury. PLoS One. 2016;11:e0159872.

14. Shinohara, K., Liu, X., Morgan, D.A., Davis, D.R., Sequeira-Lopez, M.L.S., Cassell, M.D., Grobe, J.L., Rahmouni, K., Sigmund, C.D.

10. Littlejohn, N.K., Keen, H.L., Weidemann, B.J., Claflin, K.E., Tobin, K.V., Markan, K.R., Park, S., Naber, M.C., Gourronc, F.A., Pearson,

Selective Deletion of the Brain-Specific Isoform of Renin Causes Neurogenic Hypertension. Hypertension 68: 1385-1392, 2016

N.A., Liu, X., Morgan, D.A., Klingelhutz, A.J., Potthoff, M.J., Rahmouni, K., Sigmund, C.D., Grobe, J.L. Suppression of Resting Metabolism by the Angiotensin AT2 Receptor. Cell Reports 16: 1548-1560, 2016.

15. Agbor, L.N., Ibeawuchi, S.C., Hu, C., Wu, J., Davis, D.R., Keen, H.L., Quelle, F.W., Sigmund, C.D. Cullin-3 Mutation Causes Arterial Stiffness and Hypertension Through a Vascular

11. Wu, J., Sigmund, C.D. Hypertension: A Disease that Strikes Around

Smooth Muscle Mechanism. JCI Insight 1:e91015, 2016.

the Clock (Editorial Commentary). Hypertension 67: 493-495, 2016.

12. Stump, M., Guo, D-F., Lu, K-T, Mukohda, M., Cassell, M.D., Norris, A.W., Rahmouni, K., and Sigmund, C.D. Nervous System Expression of PPARg and Mutant PPARg Has Profound Effects on Metabolic Regulation and Brain Development. Endocrinology 157: 4266-4275, 2016.

13. Prasad, A.M., Ketsawatsomkron, P., Nuno, D.W., Koval, O.M., Dibbern, M.E., Venema, A.N., Sigmund, C.D., Lamping, K.G.,

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MICHAEL J. WELSH, MD Dr. Michael Welsh’s research continues to

ROY J. CARVER CHAIR IN INTERNAL MEDICINE DIRECTOR, PAPAJOHN BIOMEDICAL INSTITUTE PROFESSOR, DEPARTMENT OF INTERNAL MEDICINE

focus on understanding how the inherited disease cystic fibrosis (CF) causes lung destruction and on the development of new treatments.

PROFESSOR, DEPARTMENT OF MOLECULAR PHYSIOLOGY AND BIOPHYSICS

Cystic fibrosis is an inherited disease that predisposes people to

PROFESSOR, DEPARTMENT OF NEUROSURGERY

lung infections. The infections cause inflammation, progressive

INVESTIGATOR, HOWARD HUGHES

destruction of lung tissue, and eventually respiratory failure.

MEDICAL INSTITUTE

A healthy respiratory system mounts a formidable defense against bacteria. In people with CF, that shield breaks down. Understanding how CF disrupts the defensive shield is a major goal of the Welsh lab. In the past, a critical roadblock to solving this puzzle was lack of an animal that develops disease like human CF. Unfortunately, the common lab animal model, mice, remain healthy when they have a defective CF gene. Thus, their utility for investigating CF has been limited. To circumvent this roadblock, Dr. Welsh and his colleagues engineered pigs with a mutated CF gene. The CF pigs develop lung disease that mimics the disease in humans. By studying CF pigs, they discovered that the lung’s shield against bacterial infections is impaired at the point where bacteria first encounter the lung, the surface of the tracheal and bronchial passages. There is a thin layer of liquid and mucus covering the breathing passages. That thin layer of liquid and mucus kills bacteria and removes them from the lung. In CF, Welsh and his colleagues discovered that the liquid is abnormally acidic, and the acidity disrupts the defensive shield. With this knowledge, Welsh and his colleagues returned to mice, asking why are lung defenses not disrupted in CF mice? They discovered that in contrast to pigs and humans with CF, the thin layer of liquid on the breathing passages of CF mice is not abnormally acidic. As a result, their defenses against bacteria remain intact.

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That finding immediately raised the question, why is lung liquid not acidic in CF mice? Welsh and his colleagues discovered that the air passages in humans and pigs contain a proton pump that secretes acid into the airway liquid. The acid secretion is counteracted by secretion of base (bicarbonate), thereby maintaining a normal acid-base balance. In pigs and humans, mutation of the CF gene eliminates base secretion. As a result, acid secretion is unchecked, airway liquid becomes acidic, and defenses are impaired. In contrast to pig and human lungs, mouse lungs lack the proton pump. Thus, even though mutating the CF gene eliminates base secretion in mice, because they lack the proton pump, the airway liquid does not acidify. Hence, the shield against bacteria remains intact. This discovery explains a decades old mystery about why CF mice do not develop lung disease. It underscores the importance of acid-base balance for lung health. And it nominates a new proton pump as a potential therapeutic target for treating CF. These discoveries will hopefully lead physicians and scientists to develop new preventions and treatments. Personal accomplishments This year, Dr. Welsh delivered the University of Utah Annual Medical Research Day Keynote Lecture. For 2017, he has been invited to deliver the Keynote Address at the American Thoracic Society International Conference in Washington, D.C. The American Physiological Society will also give him the Walter B. Cannon Award. In addition, he learned that he will receive the Steven C. Beering Award from the Indiana University School of Medicine. Companies co-founded In 2016, Dr. Welsh co-founded Talee Bio, Inc. a biotech company focused on gene therapy, initially for cystic fibrosis. He previously founded Emmyon, Inc. and Exemplar Genetics, which was acquired by Intrexon.

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PUBLICATIONS

2016 publications related to this research:

1. Hisert KB, Schoenfelt KQ , Cooke G, Grogan B, Launspach JL, Gallager CG, Donnelly SC, McKone EF, Welsh MJ, Singh PK, and Becker L. Ivacaftor-induced proteomic changes suggest monocyte defects may contribute to the pathogenesis of Cystic Fibrosis. Am. J. Respir. Cell Mol. Biol., 54:594-597, 2016. PMCID: PMC 4821059.

2. Meyerholz DK, Lambertz AM, Reznikov LR, Ofori-Amanfo GK, Karp PH, McCray PB, Welsh MJ, and Stoltz DA. Immunohistochemical detection of markers for translational studies of lung disease in pigs and humans. Toxicologic Pathology, 44:434-441, 2016. PMCID: PMC4805467.

3. Li X, Tang XX, Vargas Bounfiglio LG, Comellas AP, Thornell IM, Ramachandran S, Karp PH, Taft PJ, Sheets K, About Alaiwa MH, Welsh MJ, Meyerholz DK, Stoltz DA, and Zabner J. Electrolyte transport properties in distal small airways from cystic fibrosis pigs with implications for host defense. Am J Physiol Lung Cell Mol Physiol. 310:L670-679, 2016. PMCID: PMC4824164.

4. Shah VS, Ernst S, Tang XX, Karp PH, Parker CP, Ostedgaard LS, Welsh MJ. Relationships Among CFTR Expression, HCO3- Secretion, and Host Defense May Inform Gene- and Cell-Based Cystic Fibrosis Therapies. Proc Natl Acad Sci USA. 113(19):5382-5387, 2016. PMCID: PMC4868420.

5. Bartlett JA, Ramachandran S, Wohlford-Lenane CL, Barker CK, Pezzulo AA, Zabner J, Welsh MJ, Meyerholz DK, Stoltz DA, and McCray PB Jr. Newborn Cystic Fibrosis Pigs Have a Blunted Early Response to an Inflammatory Stimulus. Am. J. Respir. Crit. Care Med., 194:845-854, 2016. PMCID: PMC5074652.

6. Tang XX, Ostedgaard LS, Hoegger MJ, Moninger TO, Karp PH, McMenimen JD, Choudhury B, Varki A, Stoltz DA, and Welsh MJ. Acidic pH Increases Airway Surface Liquid Viscosity in Cystic Fibrosis. J. Clin. Invest., 126:879-891, 2016. PMCID: PMC4767348.

7. Adam RJ, Hisert KB, Dodd JD, Grogan B, Launspach JL, Barnes JK, Gallagher CG, Sieren JP, Gross TJ, Fischer AJ, Cavanaugh JE, Hoffman EA, Singh PK, Welsh MJ, McKone EF, Stoltz DA. Acute Administration of Ivacaftor to People with Cystic Fibrosis and a

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

MICHAEL J. WELSH, MD Balance Forward

$28,813

Revenue Disbursements to UI Total Revenue

$69,301

Equipment (2.3%) Personnel (12.9%)

Expenses Personnel $469 Lab Supplies $3,074 Equipment $84 Total Expenses $3,627 Ending Balance

$94,488

G551D-CFTR Mutation Reveals Smooth Muscle Abnormalities. JCI Insight. 2016; 1(4):e86183. PMCID: PMC4855508.

Lab Supplies (84.8%)

11. Steines B, Dickey DD, Bergen J, Excoffon KJDA, Weinstein JR, Li X, Yan Z, Abou Alaiwa MH, Shah VS, Bouzek DC, Powers LS, Gansemer ND, Ostedgaard LS, Engelhardt JF, Stoltz DA, Welsh MJ, Sinn PL, Schaffer

8. Shah VS, Meyerholz DK, Tang XX,Reznikov L, Abou Alaiwa M, Ernst SE, Karp PH, Wohlford-Lenane CL, Heilmann KP, Leidinger MR, Allen

DV, Zabner J. CFTR gene transfer with AAV improves early cystic fibrosis pig phenotypes. JCI Insight, 1(14):e88728, 2016. PMCID: PMC5033908.

PD, Zabner J, McCray PB Jr, Ostedgaard LS, Stoltz DA, Randak CO, and Welsh MJ. Airway Acidification Initiates Host Defense Abnormalities in Cystic Fibrosis Mice. Science, 351:503-507, 2016. PMCID: PMC4852973.

12. Cooney AL, Abou Alaiwa MH, Shah VS, Bouzek DC, Stroik MR, Powers LS, Gansemer ND, Meyerholz DK, Welsh MJ, Stoltz DA, Sinn PL, McCray PB Jr. Lentiviral-mediated phenotypic correction of cystic

9. Abou Alaiwa MH, Launspach JL, Sheets KA, Rivera JA,

fibrosis pigs. JCI Insight, 1(14):e88730, 2016. PMCID: PMC5027966.

Gansemer ND, Taft PJ, Thorne PS, Welsh MJ, Stoltz DA, Zabner J. Repurposing Tromethamine as Inhaled Therapy to Treat CF Airway Disease. JCI Insight. 2016; 1(8). pii: e87535.

10. Reznikov LR, Meyerholz DK, Adam RJ, Abou Alaiwa M, Jaffer O, Michalski AS, Powers LS, Price MP, Stoltz DA, Welsh MJ. Acid-Sensing Ion Channel 1a Contributes to Airway Hyperreactivity in Mice. PLOS ONE, 11(11): e0166089. doi:10.1371/journal.pone.0166089. PMCID: PMC5098826.

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JOHN WEMMIE, MD, PHD Dr. John Wemmie’s research focuses on

ROY J. CARVER CHAIR IN PSYCHIATRY AND NEUROSCIENCE

brain pH (acidity) and a family of ion

PROFESSOR, DEPARTMENT OF PSYCHIATRY

channels play an important role in behavior

PROFESSOR, DEPARTMENT OF MOLECULAR

channels in the brain that are activated by acid called acid-sensing ion channels (ASICs). Previous studies indicate these

PHYSIOLOGY AND BIOPHYSICS PROFESSOR, DEPARTMENT OF NEUROSURGERY

and brain function in mice and contribute to plasticity at synapses, points of chemical communication between neurons.

Dr. Wemmie was only recently named Roy J. Carver Chair and has therefore had less time to capitalize on this important addition to his research. He is planning to use the Carver funds to purchase equipment and hire personnel to advance elements of his work considered higher in risk and also of greater potential reward. There are several projects on-going in Dr. Wemmie’s laboratory that will benefit greatly from Carver funding. Three of these are outlined below: 1. Acid-sensing ion channels (ASICs) in drug addiction. Recent work in the Wemmie lab suggests that ASICs play an important role in the self-administration of cocaine and other drugs, including opioids, as well as in the structure and function of key synapses in the brain underlying drug-seeking behaviors. Dr. Wemmie plans to use the Carver funds to test whether a pH-regulating enzyme, carbonic anhydrase, which can influence the action of ASICs at synapses, might be targeted by drugs to regulate ASIC activity in humans and thus attenuate addiction. There are already a number of drugs that affect carbonic anhydrase already approved for use in humans for other illnesses, and thus might be readily available to treat people with addictive disorders.

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2. ASICs in alcohol addiction. Alcohol is abused by more

Dr. Wemmie accomplished several important goals this

people than cocaine and opioids, and alcohol abuse

year. Because one of his RO1 grants will be completed in

can exact an even greater cost on the health and

February 2017, it is critical for him to obtain additional

well-being of individuals and society. Because alcohol

outside funding in order to maintain a cutting edge research

abuse is thought to share mechanisms with other drugs

program. Therefore obtaining additional grant funding was

of abuse, it will be valuable to know whether ASICs

the top priority for 2016. Dr. Wemmie prepared, wrote, and

may also contribute to alcohol abuse and whether

submitted three distinct RO1 grants to the NIH in 2016. The

ASICs might be therapeutically targeted to treat

first application focused on the amygdala and panic disorder-

alcohol use disorders. Dr. Wemmie plans to start by

related physiology and behavior in mice. The second focused

manipulating ASICs in mice and test the effects on

on a novel brain imaging strategy that is sensitive to pH. The

alcohol-seeking behaviors. Because testing alcohol

third focused on investigating abnormal pH and circuitry in

is more challenging than cocaine and opioids in

the cerebellum of patients with bipolar disorder. The first

laboratory animals, the Carver funds will be helpful for

two applications received scores of 11 and 16 percentile and

supporting this endeavor and for gathering preliminary

have a good chance of being funded in the current cycle.

data for future grant applications on this project.

The third application will be reviewed this coming February. Dr. Wemmie also was the co-principal investigator on a T32

3. Novel treatments for depression. Despite currently

proposal with Dr. Peggy Nopoulos focusing on Mental Illness

available antidepressant therapies, treatment resistant

entitled “The Iowa Neuroscience Specialty Program in

depression remains a devastating problem for millions

Research Education (INSPIRE),” which received a competitive

of people. Thus, more effective treatments with distinct

score and will likely be funded starting in 2017. Dr. Wemmie

mechanisms of action are desperately needed. A new

was also a co-investigator on an NIH RO1 application led by

anti-depressant medication that is receiving increasing

Dr. Alexander Bassuk that was awarded funding in 2016.

attention is ketamine. Ketamine may have a number of advantages over current therapies. For example, it can

In 2016 Dr. Wemmie also served as President of the Psychiatry

suppress suicidal thoughts and behavior and acts much

Research Society. He served as a standing member of the

more rapidly, less than 24 hours compared to weeks for

NIH Pathological Basis of Mental Disorders and Addictions

all currently known therapies. Ketamine also produces

Study Section (PMDA). He served as a peer reviewer for

antidepressant-like effects in mice. However, ketamine

a number of publications. He participated on the search

has a number of side effects. Consequently, efforts

committee for the Director of the Neuroscience Institute,

are underway to understand how ketamine works and

which successfully recruited Dr. Ted Abel, PhD. Dr. Wemmie

to develop better ketamine-like drugs with fewer side

served as the Director of the Molecular Psychiatry Division

effects. Interestingly, Dr. Wemmie’s lab recently found

in the Department of Psychiatry. He served on the thesis

that in mice lacking ASIC1A ketamine failed to produce

committees of three PhD candidates. Dr. Wemmie wrote,

anti-depressant effects and even tended to promote

contributed to, and submitted a number of abstracts and

depression-related behavior. These unpublished findings

papers, a number of which have not yet been published.

suggest that ketamine’s effects depend on ASIC1A. They further suggest that ASIC1A may be targeted to produce effects resembling those of ketamine, and thus may offer a novel way to treat depression. Consistent with this possibility, disrupting ASIC1A or blocking its action with pharmacological antagonists produced antidepressant-like effects in mice. Dr. Wemmie’s lab is now working to better understand the mechanisms underlying this surprising interaction between ASICs and the antidepressant effects of ketamine.

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PUBLICATIONS

1. Shao W, Christensen GE, Johnson HJ, Song JH, Durumeric OC, Johnson CP, Shaffer JJ, Magnotta VA, Fiedorowicz JG, Wemmie JA. Population Shape Collapse in Large Deformation Registration of MR Brain Images. 7th International Workshop on Biomedical Image Registration. 2016.

2. Du J, Price MP, Taugher RJ, Grigsby D, Ash JJ, Stark AC, Wemmie JA, Welsh MJ. Transient Acidosis while Retrieving a FearRelated Memory Enhances Its Lability. e-Life. Under Revision

3. Gutman A, Cosme C, Worth W, Gupta S, Wemmie JA, and Ryan LaLumiere. The extinction of cocaine seeking requires a window of infralimbic activity after unreinforced lever presses. J. Neurosci. Under Revision

4. Shaffer JJ Jr., Johnson CP, Fiedorowicz JG, Christensen GE, Wemmie JA, Magnotta VA. Functional MRI Activation is Reduced in Bipolar Disorder Manic and Depressed Mood States during Flashing Checkerboard. Submitted

5. Shaffer JJ Jr., Johnson CP, Fiedorowicz JG, Christensen GE, Wemmie JA, Magnotta VA. Relationship Between Altered Functional T1Ď and BOLD Signals in Bipolar Disorder. Submitted

6. Shaffer JJ Jr., Johnson CP, Fiedorowicz JG, Christensen GE, Magnotta VA, Wemmie JA. Quantitative T1Ď Mapping Detects Abnormalities of the Cerebellum and Basal Ganglia in Mood States of Bipolar Disorder. In preparation.

7. Taugher RJ, Kreple CJ, Dlouhy BJ, Ghobbeh A, Sowers LP, Wang Y, Naryanan K, LaLumiere R, Wemmie JA. The amygdala and dorsal periaqueductal gray in CO2-evoked behavior. In preparation.

8. Faraci FM., Taugher RJ, Lynch C, Fan R, Gupta S, and Wemmie JA. Acid-Sensing Ion Channels: Novel Mediators of Cerebrovascular Responses. In preparation.

9. Taugher RJ, Lu Y, Fan R, Wang Y, Kreple CJ, Wemmie JA. Neuronal ASIC1A is critical for fear-related behavior. In preparation.

10. Ghobbeh A, Alam S, Taugher RJ, Fan R, LaLumiere R, Wemmie JA. The role of acid-sensing ion channels in instrumental learning. In preparation.

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

JOHN WEMMIE, MD, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$22,500

Travel (100%)

$0

Expenses Travel $1,986 Total Expenses Ending Balance

$1,986 $ 20,514

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ROBERT C. PIPER, PHD Several cellular activities are performed

ROY J. CARVER PROFESSOR IN MOLECULAR PHYSIOLOGY AND BIOPHYSICS DIRECTOR, COLLEGE OF MEDICINE CORE RESEARCH FACILITIES PROFESSOR, DEPARTMENT OF MOLECULAR PHYSIOLOGY AND BIOPHYSICS PROFESSOR, DEPARTMENT OF INTERNAL MEDICINE

by proteins embedded in the cell’s surface membrane. Dr. Robert Piper’s research aims to understand how these proteins are degraded.

Funds from the Roy J. Carver Charitable Trust were used in 2016 to help establish a new cell model system in the laboratory to study how proteins move into lysosomes. The model system consists of human haploid cells, and Carver Trust funds were used to underwrite efforts to establish methods to knockout, alter, or replace genes within them. The Piper lab studies how membrane proteins are moved from the cell surface into lysosomes where they undergo degradation. Degradation of membrane proteins is important because it serves as a major way to regulate their activity. An example are so-called growth factor receptors, that reside in the surface membrane and detect a variety of growth hormones in the bloodstream and signal for cells to grow and divide. Normally, cells destroy these receptors after they signal, providing a way to limit the cell’s response to growth hormones. Without the ability to destroy those receptors, the stimulus for cell growth can go unchecked and can contribute to pathologies such as cancer. Control of this destruction process is conveyed by the attachment of ubiquitin to the membrane proteins destined for degradation. After attachment, ubiquitin is recognized by a host of cell machinery that ultimately moves the modified proteins into the interior of lysosomes, where they undergo degradation. To study this process at the cellular and molecular level, the Piper lab has used a combination of protein structure techniques (eg. NMR and protein crystallography) combined with genetic analysis in model eukaryotic cell systems such as yeast. This has provided the basic mechanistic framework for how biologists generally understand how membrane proteins are degraded in lysosomes. Exactly how this basic framework operates and how it is regulated in human cells remains

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largely unknown, partly because the definitive genetic tools

partners to a variety of proteins that serve critical cellular

available in yeast (such as precise knockout and replacement of

functions. In addition, this workflow has been implemented in

particular genes) have not been available in mammalian cells.

undergraduate classes as a way to teach modern genetics and molecular biology. Development of this class was supported

New tools have now been developed to make specific changes

by an NSF grant and implementation of the class has spread

in the DNA of mammalian cells, offering the potential for

from its initial effort through at the University of Iowa to now

a new era of highly precise and robust experimentation in

include undergraduate classes at three other Midwest colleges.

human cells. In addition, a new cell model in the form of cultured human haploid cells has been developed, offering

Another accomplishment centered on a fundamental cell

a far more effective way to obtain derivative experimental

biological question. It is known that membrane proteins

cell lines that have a specifically altered genome. These tools

embedded in the cell’s surface membrane are taken up

have potential to provide an experimental system that could

(internalized) into the interior of cells as a means to regulate

answer many of the questions surrounding how human cells

their activity. Often, these membrane proteins are returned

execute and regulate the degradation of cell surface proteins.

to the cell surface in a process called “recycling.” While much is known about the machinery that mediates internalization,

Funds from the Carver Trust were used to help the Piper lab

hardly anything is known about the process of recycling,

develop and characterize such an experimental system. This

which plays an outsized role in controlling a wide variety

was done by characterizing haploid cells at the cell biological

of cell functions. The Piper lab has now completed a novel

and biochemical level that established the organization of

genetic screen that has uncovered dozens of new proteins that

the membrane structures within the cells and the protein

play central roles in the process of recycling. Many of these

machinery that controls the activity of these structures.

proteins are conserved from yeast to humans and many were

Techniques were also established and developed to knock

previously not recognized to have a known function or to

out genes of interest within these cells and to replace genes

function in recycling. Unexpectedly, these studies uncovered a

with altered versions. Finally, bioinformatics tools were

link between cellular metabolism and recycling of membrane

implemented that allow monitoring of gene expression

proteins, that has implications for cancer and diabetes.

changes as well as thoroughly defining the genome of each experimental cell line developed in the future. One major accomplishment was the completion of a new system for finding binding partners for cellular proteins. Much of a protein’s function is mediated by its ability to interact or bind to other proteins. In addition, many times the reason a disease variant of a protein does not work properly is because it is unable to correctly interact with a subset of its binding partners. The Piper lab has now developed a new way to discover the repertoire of binding partners that protein has and compare how that repertoire might change for a disease-causing variant. This method uses an established technique using yeast genetics, but pushes it to be far more comprehensive and quantitative through the application of next-generation high throughput DNA sequencing and the development of custom software tools that are specifically tailored for this process. This has lead to many collaborative studies between the Piper lab and other laboratories interested in finding functional binding

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PUBLICATIONS

1. Pashkova N, Peterson TA, Krishnamani V, Breheny P, Stamnes M, Piper RC. DEEPN as an Approach for Batch Processing of Yeast 2-Hybrid Interactions. Cell Rep. 2016 Sep 27;17(1):303-15. PMID: 27681439

2. MacDonald C, Piper RC. Cell surface recycling in yeast: mechanisms and machineries. Biochem Soc Trans. 2016 Apr 15;44(2):474-8. PMID: 27068957

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

ROBERT PIPER, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$0

Lab Supplies/ Animals (59%)

$16,875

Expenses Lab Supplies/Animals $9,767 Other $6,789 Total Expenses $16,557 Ending Balance

$318 Other (41%)

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RESEARCH PROGRAMS OF EXCELLENCE

81


DAVID F. WIEMER, PHD F. WENDELL MILLER PROFESSOR PROFESSOR, DEPARTMENT OF CHEMISTRY PROFESSOR, DEPARTMENT OF PHARMACOLOGY

The longstanding objective of Dr. David Wiemer’s program is to improve understanding of the cholesterol biosynthetic pathways and use that information to develop new compounds for cancer treatment. Pharmacological Study of Intermediates in Cholesterol Biosynthesis for Cancer Treatment Over the past year, funds from the Roy J. Carver Charitable Trust have been spent on personnel, laboratory supplies and UI facilities. These investments supported preparation of a variety of new compounds as potential drugs, as well as their evaluation in biological assays that measure their impact on these important biosynthetic pathways. This current year has built upon advances of prior years and led to new strategies to modulate isoprenoid metabolism Figure 1 for potential benefits in human health. In prior years Professors Wiemer and Raymond Hohl developed many small molecules that inhibited specific enzymatic steps in these metabolic pathways selectively and observed dramatic cellular effects. For example, they have designed and synthesized molecules with activity against the central enzyme target GGDPS Figure 2. So many of those earlier compounds had a V-like shape, that it would have been easy to assume that a V-shape was necessary for activity (e.g. 1). During the past year several new families of compounds that impact the GGPP synthase target with even greater potency have been developed. One new family might be viewed as more U-shaped (2) while the other is simply linear (3). These new templates offer additional possibilities for creation of

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Figure 1. Key steps in the isoprenoid biosynthesis pathways. Shadowed boxes indicate focal points of this program including, from left to right, cholesterol regulation, the enzyme GGPP synthase (GGDPS), the phosphoantigen (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP), and the enzyme geranylgeranyl transferase II.

new, bioactive compounds. For example, the potency of

olefin with an amide group, resulting in an increase in

compound 3 was found to be enhanced in a synergistic

water solubility while still preserving biological activity at

manner when it was combined with the isomeric cisolefin 4.

nanomolar levels. Through expanded collaborations (vide infra), they also have prepared new compounds that target

This group also has advanced understanding of how the

the enzyme geranylgeranyl transferase II which might be of

natural products known as the schweinfurthins impact

benefit in treatment of multiple myeloma, and others that

regulation of cholesterol levels. In essence, they have

mimic the bacterial isoprenoid intermediate (E)-4-hydroxy-3-

determined that schweinfurthins mislead cells into recognition

methyl-but-2-enyl diphosphate (HMBPP) and may be able to

of high cholesterol levels when in fact cholesterol levels are

stimulate the human immune system to recognize malignant

lowered to the point where cell viability is impacted. Two

cells. This Carver Research Program of Excellence has

types of cancer cells, glioblastoma and chondrosarcoma,

been instrumental in providing support for the discoveries

are especially sensitive to these compounds, and thus these

that have led to these potential therapeutic advances.

materials hold promise for treatments of cancers which are nearly intractable with current therapies. New variations

This interdisciplinary research team continues to advance

on these structures included replacement of the central

understanding of the many roles that isoprenoids, the

R E S E A R C H P R O G R A M S O F E XC E L L E N C E

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Figure 2. General shapes of GGDPS inhibitors.

metabolic precursors for cholesterol and other important

PA. There it has been found that some of these compounds

cellular products, play in altering cancer cell growth and

have modest activity against pancreatic cancer cell lines.

modifying fundamental properties such as cancer cell migration. The dynamic interaction between chemical research

To multiply the impact of the support provided by the Carver

and medical research has built a detailed understanding of

Trust, Professor Wiemer has aggressively sought support

these complex isoprenoid biosynthesis pathways, and has

for the team from the National Institutes of Health (NIH).

been encouraged and facilitated by the support of the Roy

To date this has resulted in a four-year R01 award from

J. Carver Charitable Trust through this Research Program

NIH shared by Professors Wiemer and Holstein, as well as

of Excellence. Common ground for these laboratories has

a second four-year R01 submitted by Professors Andrew

been the science of pharmacology, the study of mechanisms

Wiemer and David Wiemer, which was funded as of April

of drug action. The combined efforts of this team have

1, 2015. These external awards magnify the impact of the

resulted in discoveries that have exceeded those otherwise

Carver Trust support, reflect the high regard that external

possible from their individual research programs alone.

reviewers have for this program, and enhance the visibility and reputation of the Carver College of Medicine. They also

While Professors Wiemer and Hohl continue endeavors to

allow the PI to expand into novel areas of research that rely

control biological activity through inhibition of the central

upon manipulation of the isoprenoid biosynthetic pathways

enzyme geranylgeranyl diphosphate synthase (GGPP synthase),

to accomplish new biological objectives. During the past

two of their former students now have independent positions

year a new NIH proposal (R01) was submitted describing

of their own and have joined this effort as collaborators.

plans to create novel fluorescent prodrugs (DFW, PI; AJ

Professor Sarah A. Holstein (MD/PhD in 2004; Associate

Wiemer and ML Geng, co-PI’s). These prodrugs are designed

Professor of Oncology and Hematology at the University

to allow studies of the cellular uptake and metabolism of

of Nebraska Medical Center in Omaha) is interested in the

compounds prepared to modulate the isoprenoid biosynthetic

downstream enzyme geranylgeranyl transferase II and in

pathway. (Biological probes based on fluorescent cell

treatment of multiple myeloma. Professor Andrew J. Wiemer

cleavable protecting groups. NIH #1R01EB024510-01,

(PhD in 2008; now Assistant Professor of Medicinal Chemistry

submitted Sept. 30, 2016; proposed duration Sept. 1, 2017

at the University of Connecticut, Storrs) is interested in the

through Aug. 31,2021; requested support $1,510,003.)

early intermediates of isoprenoid biosynthesis and their ability to stimulate the immune system. Novel compounds provided to both of these research groups have resulted in significant findings over the past year, as reflected in the publications listed in this report. Finally, we also have provided analogues of the natural products known as the pawhuskins, which have structures closely related to the schweinfurthins, to researchers at the Penn State Cancer Institute in Hershey,

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

DAVID F. WIEMER, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$18,083

Other (19.8%)

$100,000

Expenses Personnel $95,797 Lab Supplies $23,352 Equipment $506 Other $13,002 Total Expenses $132,657 Ending Balance

Equipment (0.4%)

Personnel (72.2%)

Lab Supplies (17.6%)

($14,574)

PUBLICATIONS

Publications in 2016 that acknowledge the Roy J. Carver Charitable Trust

5. Mattheissen, R.A.; Wills, V.S.; Metzger, J.I.; Holstein, S.A.; Wiemer, D.F. Stereoselective Synthesis of Homoneryl and Homogeranyl Triazole Bisphosphonates. J. Org. Chem. 2016, 81, 9438–9442.

1. Wiemer, A.J.; Shippy, R.R.; Kilcollins, A.M.; Li, J.; Hsiao, C.C.; Barney, R.J.; Geng, M.L.; Wiemer, D.F. Evaluation of a 7-methoxycoumarin3-carboxylic acid ester derivative as a fluorescent, cell-cleavable, phosphonate protecting group. ChemBioChem, 2016, 17, 52–55.

2. Gardner, K.D. Wiemer, D.F. Selective Prenylation of Protected Phenols for Synthesis of Pawhuskin A Analogues. J. Org. Chem. 2016, 81, 1585–1592.

3. Wills, V.S.; Zhou, X.; Allen, C.; Holstein, S.A.; Wiemer, D.F. Stereocontrolled regeneration of olefins from epoxides. Tetrahedron Lett. 2016, 57, 1335–1337.

4. Foust, B.J.; Allen, C.; Holstein, S.A.; Wiemer, D.F. A New Motif for Inhibitors of Geranylgeranyl Diphosphate Synthase. Bioorg. & Med. Chem. 2016, 24, 3734-3741.

6. Allen, C.;Kortagere, S.; Tong, H.; Mattheissen, R.A.; Metzger, J.I.; Wiemer, D.F.; Holstein, S.A. Olefin Isomers of a Triazole Bisphosphonate Synergistically Inhibit Geranylgeranyl Diphosphate Synthase. Mol. Pharm. 2016, in press (now available on line at http://molpharm.aspetjournals.org/)

7. Shippy, R.R.; Lin, X.; Agabiti, S.S.; Li, J.; Zangari, B.M.; Hsiao, C.C.; Vinogradova, O.; Wiemer, D.F.; Wiemer, A.J. Phosphinophosphonates and their tris-pivaloyloxymethyl prodrugs reveal a negatively cooperative butyrophilin activation mechanism. J. Med. Chem., 2016, accepted pending minor revisions

8. In addition, one student submitted her PhD thesis that acknowledged the Carver Trust:

9. Synthesis of Potential Opioids Based on the Natural Pawhuskins. Kevyn D. Gardner Ricossa, final defense 4/18/16.

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AMY LEE, PHD Dr. Amy Lee’s research focuses on the ASSISTANT DEAN, OFFICE OF RESEARCH

forces that regulate Ca2+ channels and

PROFESSOR, DEPARTMENT OF MOLECULAR

their roles in refining Ca2+ signals in the

PHYSIOLOGY AND BIOPHYSICS PROFESSOR, DEPARTMENT OF NEUROLOGY PROFESSOR, DEPARTMENT OF OTOLARYNGOLOGY— HEAD AND NECK SURGERY

nervous and cardiovascular systems. The lab hopes to identify new routes by which Ca2+ channels may be targeted in novel treatments for diseases associated with Ca2+ channel defects such as blindness, deafness, migraine, autism, ataxia, and cardiac arrhythmia. Dysregulation of ion channels in human disease Funds from the Roy J. Carver Charitable Trust were used to support the salaries of key personnel as well as for supplies and animal costs. Core Facilities expenses were due to use of DNA sequencing and Central Microscopy facilities. Equipment for optical and electrophysiological experiments was purchased. In addition, there were travel expenses associated with attendance of scientific meetings and visits with collaborators. In this budget period, our studies of Ca2+ channels in the retina led to 3 papers on this topic. First, we discovered a novel Ca2+ channel variant expressed in the human retina with highly unusual properties. Functional analysis of this variant revealed new molecular insights into how these channels control their opening and closing (Haeseleer et al., 2016). Second, we established unexpected roles for two Ca2+ binding proteins regulating vision by altering the physiological properties of the retinal ganglion cells (Sinha et al., 2016). Finally, we showed that an extracellular subunit of the Ca2+ channel plays a key role in organizing the structure of the photoreceptor, which can explain the visual deficits in humans bearing mutations in this protein (Kerov et al., in revision). Some of these findings formed the basis of a F31 individual NRSA grant that was awarded to Ms. Brittany Williams, a PhD student in our lab.

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Our collaboration with Dr. Sheila Baker, Assistant Professor of Biochemistry, continues to be productive. Using a CRISPR/Cas9 genome editing approach, we generated a new mouse line expressing Ca2+ channels bearing a mutation that renders them nonfunctional. Based on our unpublished data, we expect that photoreceptor synapses will assemble normally in these mice, thus verifying that Ca2+ channel function is dispensable for structural organization in the retina. This would be an extremely novel discovery in that synaptogenesis in the nervous system most often requires Ca2+ influx through Ca2+ channels and synaptic transmission which strengthen newly formed synapses. We expect to submit this study for publication in Science. These findings also formed the basis of a new R01 grant application which received an outstanding score (3 percent) and funding of a new NIH R21 grant with Drs. Baker and Arlene Drack in the Department of Ophthalmology and Visual Sciences to develop gene therapy approaches that could be used to correct visual defects in patients with Ca2+ mutations. Our neuronal Ca2+ channel research led to one manuscript that is in revision at the Journal of Neuroscience. The results reveal the importance of a synaptic protein, densin-180, in regulating Cav1.2 Ca2+ channels. Our study reveals a potential mechanism that may contribute to the role of Cav1.2 channel dysregulation in neuropsychiatric disease. Our work to generate more specific Ca2+ channel antibodies continues to be productive, resulting in two collaborative publications (Stanika et al., 2016 and Zhu et al., 2017).

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PUBLICATIONS

1. Haeseleer, F., Williams, B., and Lee, A. (2016) Characterization of C-terminal splice variants of Cav1.4 Ca2+ Channels in human retina. J. Biol. Chem., 291(30):15663-73

2. Krueger, J.N., Moore, S.J., Parent, R., Lee, A., and G.G. Murphy. (2016) A novel mouse model of the aged brain: Over-expression of the L-type voltage-gated calcium channel CaV1.3. Behav. Brain Res., pii: S0166-4328(16)30418-1

3. Stanika, R., Campiglio, M., Pinggera, A., Lee, A., Striessnig, J., Flucher, B., and G. Obermair. (2016) Splice variants of the Cav1.3 L-type calcium channel regulate dendritic spine morphology. Sci. Reports 6:34528

4. Sinha, R., Lee, A., Rieke, F., and F. Haeseleer. (2016) Lack of CaBP1/caldendrin or CaBP2 leads to altered ganglion cell responses. eNeuro 3(5). pii: ENEURO.0099-16.2016

5. Zhu, L., Almaça, J., Dadi, P.K., Hong, H., Sakamoto, W., Rossi, M., Lee, R.J., Vierra, N.C., Lu, H., Cui, Y., McMillin, S.M., Perry, N.A., Gurevich, V.V., Lee, A., Kuo, B., Leapman, R.D., Matschinsky, F.M., Doliba, N.M., Urs, N.M., Caron, M.G., Jacobson, D.A., Caicedo, A., and J. Wess. β -Arrestin-2 is an essential regulator of pancreatic β-cell function under physiological and pathophysiological conditions. Nat. Commun. (in press)

6. Wang, S., Hagen, J., Hardie, J., Stanika, R.I., Obermair, G.J., Kennedy, M.B., Colbran, R.J., and Lee, A. Densin-180 controls the trafficking and signaling of voltage-gated Cav1.2 Ca2+ channels at excitatory synapses J. Neurosci. (in revision)

7. Kerov, V., Soh, D., Joiner, M., Wiliams, B., Gutierrez, W., Laird, J., Yoshimatsu, T., Bhattarai, S., Artemyev, N.O., Drack, A.V., Wong, R.O., Baker, S.A., and Lee, A. a2d-4 maintains the presynaptic density of voltage-gated Cav1.4 Ca2+ channels and the organization of the photoreceptor ribbon synapse J. Neurosci. (in revision).

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

AMY LEE, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$50,314

Equipment (3.8%)

Equipment Maintenance (.1%)

$200,000 Other (16.3%)

Expenses Personnel $105,745 Lab Supplies $28,575 Equipment $6,435 Equipment Maintenance $59 Other $27,502 Total Expenses $168,317 Ending Balance

Lab Supplies (17%)

Personnel (62.8%)

$81,997

GRANTS ACQUIRED

1. NIH R21 EY027054 Rescue of photoreceptor synapses

2. Role: Co-I, $275,000 direct costs, 08/01/16-07/31/18

3. NIH R01 EY026817 Calcium channels in retinal photoreceptors

4. Role: PI, $2,001,716 direct costs (Score = 15, 3%)

5. NIH F31 EY026477, Characterization of a Human Cav1.4 Splice Variant (PI: Brittany Williams), Role: Sponsor, 06/17/16-06/16/18

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VAL C.

EDWIN

SHEFFIELD, MD, PHD

STONE, MD, PHD

ROY J. CARVER CHAIR IN MOLECULAR GENETICS

SEAMANS-HAUSER CHAIR IN

DIRECTOR, DIVISION OF MEDICAL GENETICS DIRECTOR, INTERDEPARTMENTAL RESEARCH

MOLECULAR OPHTHALMOLOGY

PROGRAM IN HUMAN MOLECULAR GENETICS PROFESSOR, DEPARTMENT OF PEDIATRICS, MEDICAL GENETICS PROFESSOR, DEPARTMENT OF

DIRECTOR, WYNN INSTITUTE FOR VISION RESEARCH DIRECTOR, CARVER FAMILY CENTER FOR MACULAR

DEGENERATION PROFESSOR, DEPARTMENT OF OPHTHALMOLOGY AND VISUAL SCIENCES

OPHTHALMOLOGY AND VISUAL SCIENCES

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The research of Drs. Val Sheffield and Ed Stone focuses on identification of genes causing inherited eye diseases, the understanding of the normal function of these genes, as well as the mechanisms

5. utilizing animal models and induced pluripotent stem cell (iPSCs) to develop interventions and treatments; and 6. implementing treatments for human eye diseases. Carver Trust funds were spent on continued development of mouse models of Bardet-Biedl syndrome (BBS), a syndromic

by which mutations in these genes lead to

form of inherited blindness, in order to better understand

disease. The ultimate goal of their research

the mechanisms involved in BBS blindness, as well as other

is the prevention and treatment of inherited eye diseases.

BBS associated disorders including obesity and diabetes. Novel mouse models were created in the past year that allow for the study of optimum timing for treatment to prevent BBS associated blindness. Carver Trust funds were also spent on the continued study of a mouse glaucoma model

Gene-Directed Therapy of Inherited Eye Diseases

(Myocillin mouse) and the application of genome editing using the CRISPR-Cas9 system for treatment of human

Funds from the Roy J. Carver Charitable Trust were used in

glaucoma initiated by utilizing this mouse model. Specifically,

2016 to purchase laboratory supplies needed to search for

attempts to alter the mutation causing the disease within the

genes involved in genetic eye diseases and to develop and

mouse genome were successfully performed. This strategy

maintain mouse models of eye diseases, which are being used

prevents the disease mechanisms from occuring by altering

to develop treatments. Some of the funds were used to support

the very basis of the disease at the level of the gene in the

personnel working toward the overall goal and to identify and

specific cells of the eye that are involved in development of

enroll patients into the program’s human studies.

glaucoma. This is a challenging approach, which has the

The overall goal of the research program of Drs. Sheffield

potential of changing how inherited eye diseases are treated.

and Stone is the discovery and understanding of the processes

In addition to the use of mouse models for development

involved in human genetic eye diseases, and the utilization of

of treatments, we continue to perform experiments to

this understanding for the development of treatments. Included

determine the functions of genes that cause blindess, as well

among the diseases studied are glaucoma (a leading cause of

as experiments demonstrating interaction of different proteins

blindness) and disorders leading to retinal degeneration.

involved in blindness. These studies have the potential for

The Sheffield and Stone Carver Research Program of Excellence consists of several components, including: 1. identifying the genes and mutations that cause humans to be born blind or to go blind; 2. determining the functions of the genes and their protein products associated with blindness; 3. determining genetic and protein interactions, as well as protein complexes and networks that contribute to blindness; 4. developing animal models (primarily mouse and zebrafish models) to aid in determining how mutated genes lead to blindness;

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identifying new mechanisms involved in the disease process, with the hope that the understanding of disease mechanisms will lead to novel approaches to prevention or treatment. Progress in the past year that should be highlighted has included the identification of additional disease genes including an additional BBS gene (BBS21). Importantly, this research has continued to play a key role in elucidating the function of proteins involved in hereditary blindness, as well as the identification and characterization of protein complexes. The work aimed at developing treatment for hereditary eye diseases includes not only genome editing but also gene therapy, pharmaceuticals, and inducible pluripotent stem cells (iPS cells) for cell replacement therapy. In addition, the Stone Laboratory continues to play a key role in the implementation

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of gene therapy for Leber’s congenital amaurosis (LCA),

Beyond his research accompllishments, Dr. Sheffield’s

an early onset form of blindness, in human patients.

accomplishments in the past year include the fact that he mentors undergraduate and graduate students, the latter in

A key area in which Carver Trust funding has aided the eye

helping them obtain their PhD degrees. Four of Dr. Sheffield’s

research program is the acquisition of extramural grant

PhD students passed their PhD comprehensive exams in the

funding. Carver Trust funding allows for the acquisition of

past year. He also provides research opportunities for post-

preliminary data, which is key to the successful competition

doctoral fellows and medical students within his laboratory.

for government funding. A major accomplishment of the

He has been instumental in helping to train young faculty at

laboratories is the securing of ongoing National Institutes

the University of Iowa to help them develop research careers

of Health (NIH) funding, which has supported the

and obtain funding. Dr. Sheffield continues to serve as the

laboratories for nearly 25 consecutive years. Of note, in

Director of the Division of Medical Genetics, which provides

the past year NIH funded a P30 award to the Sheffield and

patient care to both pediatric and adult patients who have

Stone groups to provide infrastructure for the inherited eye

genetic disorders. Dr. Sheffield also serves on various advisory

disease research performed in their laboratories and the

committees. Of note, during the past year Dr. Sheffield has

laboratories of their collaborators in the Wynn Institute

served as a member of the Advisory Council for the National

for Vision Research (WIVR; www.wivr.uiowa.org).

Human Genome Research Institute, as a member of the panel of experts for the Human Health and Heredity in Africa

It should be noted that the WIVR, under the direction

program, and as an advisor to the Intellecual Disabilities and

of Dr. Stone, has submitted to the Food and Drug

Development Research Center at Boston Children’s Hospital.

Administration two applications to perform human trials using gene therapy and cell therapy to treat specific forms of inherited blindness. This is an exciting step moving beyond laboratory research to actual patient treatment. Besides his research accomplishments, Dr. Stone has many additional personal accomplishments in the past year. Of note, he continues to direct the WIVR, which has 30 faculty members who collectively supervise more than 140 people. These individuals occupy over 40,000 square feet of space distributed across eight departments and four colleges of the University of Iowa. Dr. Stone also continues to provide outstanding leadership of the non-profit genetic testing program known as the John and Marcia Carver Nonprofit Genetic Testing Laboratory (www.carverlab.org). This lab is CLIA certified and Joint Commission inspected and currently offers 24 different tests (55 different genes). In addition to the above, it should be noted that Dr. Stone is an outstanding educator. He currently mentors an MD/ PhD student in his laboratory and regularly teaches students, residents, fellows, and other faculty in the Department of Ophthalmology and Visual Sciences and elsewhere in the Carver College of Medicine. During the past year, Dr. Stone was honored by being elected to the Association of American Physicians and was one of only two new members to be invited to give a plenary lecture, which was well received.

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9. Heon E, Kim G, Qin S, Garrison JE, Tavares E, Vincent A,

PUBLICATIONS

Nuangchamnong N, Scott CA, Slusarski DC, Sheffield VC. Mutations in C80RF37 cause Bardet Biedl syndrome (BBS21). Hum Mol Genet

1. Braverman NE, Raymond GV, Rizzo WB, Moser AB, Wilkinson ME,

2016, pii:ddw096 (Epub ahead of print) PMID: 27008867

Stone EM, Steinberg SJ, Wrangler MF, Rush ET, Hacia JG, Bose M. Peroxisome biogenesis disorders in the Zellweger spectrum: an

10. Huckfeldt RM, East JS, Stone EM, Sohn EH. Phenotypic

overview of current diagnosis, clinical manifestations, and treatment

variation in a family with pseudodominant Stargardt

guidelines. Molecular Genetics and Metabolism. 2016 PMID: 26750748.

disease. JAMA Ophthalmology. 2016 PMID: 27030965.

2. Chirco KR, Whitmore SS, Wang K, Potempa LA, Halder

11. Kawasaki M, Izu Y, Hayata T, Ideno H, Niifuji A, Sheffield VC, Ezura Y,

JA, Stone EM, Tucker BA, Mullins RF. Monomeric

Noda M. Bardet-Biedl syndrome 3 regulates development of cranial base

C-reactive protein and inflammation in Age-related Macular

midline structures. Bone 2016 pii:S8756-3282(16)30043-6 PMID: 27170093

Degeneration. Journal of Pathology. 2016 PMID: 27376713.

3. Chirco KR, Sohn EH, Stone EM, Tucker BA, Mullins RF. Structural and molecular changes in the aging choroid: implications for agerelated macular degeneration. Eye (Lond). 2017 PMID: 27716747.

4. Chung DD, Frausto RF, Cervantes AE, Gee KM, Zakharevich M, Hanser EM, Stone EM, Heon E, Aldave AJ. Confirmation of the OVOL2 promotor mutation c.-307T>C in posterior polymorphous corneal dystrophy 1. PLos One. 2017 PMID:28046031.

5. Collison FT, Park JC, Fishman GA, Stone EM, McAnany JJ. Twocolor pupillometry in enhanced S-cone syndrome caused by NR2E3 mutations. Documenta Ophthalmologica. 2016 PMID: 27033713.

6. Datta P, Allamargot C, Hudson JS, Andersen EK, Bhattarai S, Drack AV, Sheffield VC, Seo S. Accumulation of non-outer segment proteins in the outer segment underlies photoreceptor degeneration in Bardet-Biedl syndrome. Proc Natl Acad Sci U S A. 2015 Jul 27 pil: 201510111 PMID 26216965

7. Guo DF, Cui H, Zhang Q , Morgan DA, Thedens DR, Nishimura D, Grobe JL, Sheffield VC, Rahmouni K. The BBSome controls energy homeostasis by mediating the transport of the leptin receptor to the plasma membrane. PLoS Genet, 2016 12(2):e1005890 PMID 26926121

8. Haziza S, Magnani R, Lan D, Keinan O, Saada A, Hershkovitz E, Yanay N, Cohen Y, Nevo Y, Houtz RL, Sheffield VC, Golan H, Parvari R. Calmodulin methyltransferase is required for growth, muscle strength, somottosensory development and brain function. PLoS Genet 2015 6;11(8):e1005388 PMID 26247364

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12. Lu W, Hu H, Sevigny J, Gabelt BT, Kaufman PL, Johnson EC, Morrison JC, Zode GS, Sheffield VC, Zhang X, Laties AM, Mitchell CH. Rat, mouse, and primate models of chronic glaucoma show sustained elevation of extracellular ATP and altered purinergic signaling in the posterior eye. Invest Ophthalmol Vis Sci 2015 1;56(5):3075-83. PMID 260024091

13. McAnany JJ, Park JC, Collison FT, Fishman GA, Stone EM. Abnormal 8-Hz flicker electroretinograms in carriers of X-linked retinoschisis. Documenta Ophthalmologica. 2016 PMID: 27369766.

14. Muhammad E, Levitas A, Singh SR, Braiman A, Ofir R, Etzion S, Sheffield VC, Etzion Y, Carrier L, Parvari R. PLEXHM2 mutation leads to abnormal localization of lysosomes, impaired autophagy flux and associates with recessive dilated cardiomyopathy and left ventricular noncompaction. Hum Mol Genet 2015 [Epub ahead of print] PMID: 26464484

15. Preist JR, Osoegawa K, Mohammed N, Nanda V, Kundu R, Schultz K, Lammer EJ, Girirajan S, Scheetz T, Waggott D, Haddad F, Reddy S, Bernstein D, Burns T, Steimle JD, Yang XH, Moskowitz IP, Hurles M, Lifton RP, Nickerson D, Barnshad M, Eichier EE, Mital S, Sheffield V, Quertemous T, Gelb BD, Portman M, Ashley EA. De Novo and rare variants at multiple loci support the oligogenic origins of atrioventricular septal heart defects. PLos Genet 2016 12(40:e2005963 doi: 10.1371/journal.pgen eCollection PMIC 27058611

16. Reish O, Aspit L, Zouella A, Roth Y, Polak-Charcon S, Baboushkin T, Benyamini L, Scheetz TE, Mussaffi H, Sheffield VC, Parvari R. A homozygous Nme7 mutation is associated with situe inversus totalis. Hum Mat 2016 37(8):727-31 PMID 27060491

17. Scheetz TE, Roos BR, Solivan-Timpe F, Miller K, DeLuca AP, Stone EM, Kwon YH, Alward WL, Wang K, Fingert JH. SQSTM1 mutations and glaucoma. PLoS One. 2016 PMID: 27275741.

18. Shankar SP, Hughbanks-Wheaton DK, Birth DG, Sullivan LS, Conneely KN, Bowne SJ, Stone EM, Daiger SP. Autosomal dominant retinal dystrophies caused by a founder splice site mutation, c.828+3A.T in PRPH2 and protein haplotypes in trans as modifiers. Investigative Ophthalmology and Visual Science. 2016 PMID: 26842753.

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

VAL SHEFFIELD, MD, PHD & EDWIN STONE, MD, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$156,333

$200,000

Expenses Personnel $90,466 Lab Supplies/Animals $80,033 Travel $3,965 Equipment $3,089 Equipment Maintenance $13,310 Other $4,319 Total Expenses $195,182 Ending Balance

Other (2.2%)

Personnel (46.3%)

Equipment Maintenance (6.8%)

Equipment (1.6%)

Travel(2%)

Lab Supplies/ Animals (41%)

$161,151

19. Starks RD, Beyer AM, Guo DF, Boland L, Zhang Q ,

23. Wiley LA, Burnight ER, Drack AV, Banach BB, Ochoa D, Cranston

Sheffield VC, Rahmouni K. Regulation of insulin receptor

CM, Madumba RA, East JS, Mullins RF, Stone EM, Tucker BA. Using

trafficking by Bardet-Biedl Syndrome proteins. PLoS Genet

patient-specific induced pluripotent stem cells and wild-type mice to

2015 23;11(6):e1005311 doi:10.1371 PMID: 26103456

develop a gene augmentation-based strategy to treat CLN3-associated retinal degeneration. Human Gene Therapy. 2016 PMID: 27400765.

20. Weed MC, Almeida DR, Chin EK, Stone EM. Distinguishing optic pathway glioma and retinitis pigmentosa with visual field testing. Canadian Journal of Ophthalmology. 2016 PMID: 27316291.

24. Zhu W, Gramlich OW, Laboissonniere L, Jain A, Sheffield VC, Trimarchi JM, Tucker BA, Kuehn MH. Transplantation of iPSCderived TM cells rescues glaucoma phenotypes in vivo. Proc Natl

21. Wiley LA, Beebe DC, Mullins RF, Stone EM, Tucker BA. A method for

Adac Sci U S A 2016 21;113(25):E3492-500 PMID 27274060

sectioning and immunohistochemical analysis of stem cell-derived 3-D organoids. Current Protocols in Stem Cell Biology. 2016 PMID: 27171793.

22. Wiley LA, Burnight ER, DeLuca AP, Anfinson KR, Cranston CM, Kaalberg EE, Penticoff JA, Affatigato LM, Mullins RF, Stone EM, Tucker BA. cGMP production of patient-specific iPSCs and photoreceptor precursor cells to treat retinal degenerative blindness. Scientific Reports. 2016 PMID: 27471043.

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CURT D. SIGMUND, PHD Dr. Curt Sigmund’s research uses state-

ROY J. CARVER CHAIR IN HYPERTENSION

of-the-art molecular genetic techniques

CHAIR AND DEPARTMENT EXECUTIVE OFFICER,

physiological pathways regulating blood

DEPARTMENT OF PHARMACOLOGY PROFESSOR, DEPARTMENT OF PHARMACOLOGY PROFESSOR, DEPARTMENT OF MOLECULAR

to alter the mouse genome to uncover the molecular, genetic, cellular, and

pressure and when they go awry— hypertension.

PHYSIOLOGY & BIOPHYSICS PROFESSOR, DEPARTMENT OF INTERNAL MEDICINCE

Functional Genomics of Cardiovascular Disease In 2016, funds from the Roy J. Carver Charitable Trust (>$150,000) supporting the RPOE in Functional Genomics of Cardiovascular Disease were used to: 1) support salary for a research scientist; 2) support salary of a postdoctoral fellow; 3) provide laboratory supplies; 4) purchase and maintain experimental mice; 5) travel to a scientific meeting for Dr. Sigmund and members of his research laboratory; 6) repair of equipment; 7) purchase of new software and equipment; 8) costs of scientific publication; and 9) use of UI Core Research Facilities. As indicated in last year’s report, some of the funds to upgrade several instruments used to measure vascular reactivity, to upgrade equipment used to measure arterial blood pressure in mice, and to purchase new equipment for the measurement of vascular stiffness, an index of vascular disease. The aquisition of this equipment would have not been possible without support from the Carver Trust. This has been a particularly productive year both in terms of research results leading to publications and for acquisition of new research grants. First, the National Institutes of Health grant application entitled “Role of PPARg and the PPARg Target Gene RBP7 in the Endothelium” was funded on April 1, 2016, for four years at a total budget of $2,512,664. Funds from both the Carver Chair and RPOE were essential for the acquisition of preliminary data supporting this grant. With this NIH R01 grant funded, Dr. Sigmund’s portfolio of funding now includes the following:

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NIH NHLBI R01 “Hypertension: Role of

journal Cell Reports entitled “Suppression of Resting

Smooth Muscle Cullin-3 and the CRL3

Metabolism by the Angiotensin AT2 Receptor.” This will

Complex” (funded through March 2019)

likely be considered a seminal paper as it defines for the first time a cross talk between renin-angiotensin action in

NIH NHLBI R01 “Role of PPARg and the PPARg Target

the brain with its action in adipose (fat) tissue. This study

Gene RBP7 in the Endothelium” (funded through March 2020)

has important implications for hypertension in obesity and further evidences collaboration and synergy in the program.

NIH NHLBI P01 (PPG) “Genetic and Signaling Mechanisms in the Central Regulation of Blood

In preparation for the competitive renewal of Dr. Sigmund’s

Pressure” (funded through March 2018)

PPG, we have assembled a working group consisting of Dr. Sigmund (overall PI and leader of Project 2), Dr. Rahmouni

American Heart Association Strategically Focused

(leader of Project 3), Dr. Mark (leader of Project 1), Dr.

Hypertension Research Network (funded through March 2019)

Grobe (co-investigator of Project 2), Dr. Cui (new faculty hired in the Department of Pharmacology by Dr. Sigmund),

In aggregate this represents over $15 million in research

and Dr. Kwitek (Associate Director of the Iowa Institute for

support to the Sigmund laboratory and UI Hospitals

Human Genetics). This group has been meeting regularly

and Clinics Center for Hypertension Research.

to define the projects in the competitive renewal. This will be very challenging for several reasons. First, the number

As indicated in the Roy J. Carver Chair in Hypertension

of PPG applications funded by National Heart, Lung, and

Research Report, many of the expenditures over the past

Blood Institute (NHLBI) has steadily decreased over the

year and planned for next year revolve around preparations

past 2 years. Second, budget uncertainty has made the

to renew Dr. Sigmund’s Program Project Grant entitled

NHLBI very cautious in awarding large grants such as

“Genetic and Signaling Mechanisms in the Central

PPGs. Third, Dr. Robin Davisson (leader of Project 1) and

Regulation of Blood Pressure” which enters its final year

Professor at Weill Cornel Medical School has left Cornell

of current funding on April 1, 2017. Some of the research

and is no longer conducting experimental research. Thus,

progress on the PPG is detailed in that report. In addition,

we face the difficult task of developing a new third (and

we have been exploring other lines of research, which may

possibly fourth) project. Drs. Grobe and Cui are likely

play prominently in the competitive renewal of that grant

participants as both are highly innovative NIH R01-funded

program. For example, by taking advantage of unique and

investigators. Support of the Carver Trust through both

powerful genetically manipulated mouse models generated

the RPOE and Hypertension Chair will be instrumental in

for our NIH R01 grants, we have begun to explore the

the process of renewing this important research program.

role of PPARg and a novel signaling molecule called mTor in the brain. In collaboration with Dr. Kamal Rahmouni

Dr. Sigmund’s research and sholarship as a part of the Carver

(who is the leader of Project 3 on Dr. Sigmund’s PPG), we

RPOE and the Carver Chair in Hypertension Research has

published three papers entitled 1) “Selective Expression

resulted in several lectures and seminars in the past year

of Dominant Negative PPARg in Proopiomelanocortin

at universities and at national and international scientific

Neurons on the Control of Energy Balance,” 2) “mTORC1

meetings. Dr. Sigmund was chair of the prestigious Gordon

Signaling Contributes to Drinking but Not Blood Pressure

Research Conference on Angiotensin, held February 21-

Responses to Brain Angiotensin II,” and 3) “Nervous System

26, 2016 in Il Ciocco, Barga, Italy. He was selected as the

Expression of PPARg and Mutant PPARg Has Profound

Distinguished Lecturer in Physiological Genomics by the

Effects on Metabolic Regulation and Brain Development.”

American Physiological Society at its annual meeting, and

These papers not only serve to evidence productivity on the

gave an invited lecture on Recent Advances in Hypertension

PPG, but importantly also evidence synergy and collaboration

at the annual meeting of the American Heart Association

among the different laboratories and program leaders funded

Council on Hypertension. Dr. Sigmund presented an invited

by the program. Similarly, in a collaboration between the

seminar at University of Alabama-Birmingham in 2016 and

Sigmund and Grobe laboratories (co-investigators on Project

is scheduled to present at the University of Virginia, Weill

2 of the PPG), we published a paper in the high impact

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Cornell Medical College, Medical College of Wisconsin, and Vanderbilt University in early 2017. Dr. Sigmund will also continue his four-year term as Chair of the Publications Committee of the American Physiological Society. PUBLICATIONS

Studies in the Sigmund laboratory have resulted in 15 independent and collaborative publications in 2016:

1. Ketsawatsomkron, P., Keen, H.L., Davis, D.R., Lu, K-T., Stump, M., De Silva, T.M., Hilzendeger, A.M., Grobe, J.L., Faraci, F.M., Sigmund, C.D. A Protective Role for TIMP-4, a Novel PPARg Target Gene, in Smooth Muscle in DOCA-Salt Hypertension. Hypertension 67: 214-222, 2016.

2. Mukohda. M., Stump, M., Ketsawatsomkron, P., Hu, C., Quelle, F.W., Sigmund, C.D. Endothelial PPARg Provides Vascular Protection from IL-1β-Induced Oxidative Stress. Am J Physiol Heart Circ Physiol. 310: H39-H48, 2016.

3. Hu, C., Lu, K-T., Mukohda, M., Davis, D.R., Faraci, F.M., Sigmund, C.D. Interference with PPARg in Endothelium Accelerates Angiotensin II-induced Endothelial Dysfunction. Physiological Genomics 48: 124-134, 2016.

4. Chu, Y., Lund, D.D., Doshi, H., Keen, H.L., Knudtson, K.L., Funk, N.D., Shao, J.Q., Cheng, J., Hajj, G.P., Zimmerman, K.A., Davis, M.K., Brooks, R.M., Chapleau, M.W., Sigmund, C.D., Weiss, R.M., Heistad, D.D. Fibrotic Aortic Valve Stenosis in Hypercholesterolemic/ Hypertensive Mice. Arterioscler Thromb Vasc Biol. 36:466-474, 2016.

5. Lu, K-T., Keen, H.L., Weatherford, E.T., Sequeira-Lopez, M.L.S., Gomez, R.A., Sigmund, C.D. Estrogen Receptor Îą is Required for Maintaining Baseline Renin Expression. Hypertension 67: 992-999, 2016. (PMC4833511)

6. Stump, M., Guo, D-F., Lu, K-T., Mukohda, M., Liu, X., Rahmouni, K. Sigmund, C.D. Effect of Selective Expression of Dominant Negative PPARg in Proopiomelanocortin Neurons on the Control of Energy Balance. Physiological Genomics 48: 491-501, 2016.

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

CURT D. SIGMUND, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$82,891

Personnel (15.8%) Other (2.2%)

$100,000

Expenses Personnel $24,075 Lab Supplies/Animals $38,330 Travel $4,307 Equipment $58,735 Equipment Maintenance $3,418 Other $23,900 Total Expenses $152,765 Ending Balance

Lab Supplies/ Animals (25.1%)

Equipment Maintenance (2.2%)

Equipment (38.4%)

Travel(2.8%)

$30,126

7. Muta, K., Morgan, D.A., Grobe, J.L., Sigmund, C.D., Rahmouni, K. mTORC1 Signaling Contributes to Drinking but Not Blood Pressure

11. Wu, J., Sigmund, C.D. Hypertension: A Disease that Strikes Around the Clock (Editorial Commentary). Hypertension 67: 493-495, 2016.

Responses to Brain Angiotensin II. Endocrinology. 157: 3140-3148, 2016. 12. Stump, M., Guo, D-F., Lu, K-T, Mukohda, M., Cassell, M.D., Norris, 8. Muntner, P., Becker, R.C., Calhoun, D., Chen, D., Cowley, A.W. Jr,

A.W., Rahmouni, K., and Sigmund, C.D. Nervous System Expression

Flynn, J.T., Grobe, J.L., Kidambi, S., Kotchen, T.A., Lackland, D.T.,

of PPARg and Mutant PPARg Has Profound Effects on Metabolic

Leslie, K.K., Li,Y., Liang, M., Lloyd, A., Mattson, D.L., Mendizabal,

Regulation and Brain Development. Endocrinology 157: 4266-4275, 2016.

B., Mitsnefes, M., Nair, A., Pierce, G.L., Pollock, J.S., Safford, M.M., Santillan, M.K., Sigmund, C.D., Thomas, S.J., Urbina, E.M.

13. Prasad, A.M., Ketsawatsomkron, P., Nuno, D.W., Koval, O.M.,

Introduction to the American Heart Association’s Hypertension

Dibbern, M.E., Venema, A.N., Sigmund, C.D., Lamping, K.G.,

Strategically Focused Research Network. Hypertension. 67:674-680, 2016.

Grumbach, I.M. Role of CaMKII in Ang-II-dependent Small Artery Remodeling. Vascular Pharmacology 87: 172-179, 2016.

9. Song, K., Stuart, D., Abraham, N., Wang, F., Wang, S., Yang, T., Sigmund, C.D., Kohan, D.E., Ramkumar, N. Collecting

14. Shinohara, K., Liu, X., Morgan, D.A., Davis, D.R., Sequeira-Lopez,

Duct Renin Does Not Mediate DOCA-Salt Hypertension

M.L.S., Cassell, M.D., Grobe, J.L., Rahmouni, K., Sigmund, C.D.

or Renal Injury. PLoS One. 2016;11:e0159872.

Selective Deletion of the Brain-Specific Isoform of Renin Causes Neurogenic Hypertension. Hypertension 68: 1385-1392, 2016

10. Littlejohn, N.K., Keen, H.L., Weidemann, B.J., Claflin, K.E., Tobin, K.V., Markan, K.R., Park, S., Naber, M.C., Gourronc, F.A., Pearson,

15. Agbor, L.N., Ibeawuchi, S.C., Hu, C., Wu, J., Davis, D.R.,

N.A., Liu, X., Morgan, D.A., Klingelhutz, A.J., Potthoff, M.J., Rahmouni,

Keen, H.L., Quelle, F.W., Sigmund, C.D. Cullin-3 Mutation

K., Sigmund, C.D.*, Grobe, J.L.* Suppression of Resting Metabolism

Causes Arterial Stiffness and Hypertension Through a Vascular

by the Angiotensin AT2 Receptor. Cell Reports 16: 1548-1560, 2016.

Smooth Muscle Mechanism. JCI Insight 1:e91015, 2016.

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DOUGLAS R. SPITZ, PHD The Redox Biology and Medicine RPOE PROFESSOR, DEPARTMENT OF RADIATION

led by Dr. Douglas Spitz, focuses on

ONCOLOGY

several aspects of translational basic

DIRECTOR, FREE RADCIAL AND RADIATION BIOLOGY

science research focused on cancer

GRADUATE PROGRAM DIRECTOR, FREE RADICAL METABOLISM AND IMAGING PROGRAM

biology and therapy; taking ideas and therapy interventions from the bench

DIRECTOR, RADIATION AND FREE RADICAL

to the bedside. These include: 1) studies

to improve patient responses to cancer

RESEARCH CORE

therapy; 2) developing strategies for inhibiting normal tissue injury during cancer therapy; 3) identifying biomarkers of oxidative stress relevant to predicting outcomes following various therapies; and 4) developing image guided therapies based on changes in redox metabolism for treating cancer.

Redox Biology and Medicine Funds from the Roy J. Carver Charitable Carver trust in 2016 were used to pay salaries, purchase supplies, and pay for core lab charges for studies related to evaluating high dose vitamin C [i.e. pharmacological ascorbate that is given intravenously (IV)] and disulfiram (aka. Antabuse) to improve cancer therapy in pancreatic, brain, breast, and lung cancers. The funds also supported studies using a ketogenic diet as an intervention to improve therapy responses in head and neck cancers as well as studying the use of superoxide dismutase mimics that appear to protect normal tissues while sensitizing tumor tissues to radio-chemotherapy. Finally, funds were used for developing imaging techniques (MRI redox imaging) that may be useful in predicting responses to IV vitamin C.

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Figure 1: Median Overall Survival Comparison of RPOE supported Phase 1 Brain Cancer (GBM) Trial (NCT01752491; black lines) using Vitamin C + radiation/chemotherapy and Historical GBM Trials (blue lines). (A) Survival comparison of all 11 subjects in GBM clinical trial (NCT01752491) to Stupp et al. (2005). (B) Survival curve comparison of 8 MGMT expressing subjects in (NCT01752491) to similar subjects enrolled in Hegi et al. (Hegi et al 2005). Median overall survivals are indicated by green dots. The 95% confidence intervals are indicated (shaded area).

Collaborative interdisciplinary translational research supported

(R01CA221292; Spitz, Allen MPI; $3,356,606 total costs). In

over the first five years of the RPOE in Redox Biology

2016 this effort has also resulted in a Cancer Cell publication

and Medicine has elucidated the molecular mechanisms

very close to acceptance with many authors from the RPOE

by which intravenous high dose vitamin C selectively kills

faculty (1) as well as two more collaborative peer-reviewed

as well as sensitizes human cancer versus normal cells to

publications (2, 3). Overall, these interdisciplinary basic

conventional cancer therapies. This effort has resulted

science and clinical studies have shown that intravenous

in two completed (NCT01049880; NCT01752491), two

high dose vitamin C (aka; pharmacological ascorbate) causes

ongoing (NCT01852890; NCT02420314), and two planned

selective cancer cell toxicity and sensitization to radiation

(NCT02344355; NCT02905578) clinical trials in pancreas,

and chemotherapy by acting as a pro-oxidant in cancer

brain, and lung cancer. All these trials have shown that IV

tissues; while retaining antioxidant function in normal tissues.

high dose vitamin C is safe and well-tolerated. Furthermore,

The basic science behind this work has also uncovered

promising outcomes for patient responses to therapy have

fundamental differences in the redox biology of cancer

been obtained from all these trials. As an example, the overall

versus normal cells (1, 2) that can potentially be exploited in

survival analysis from the brain cancer trial is shown in

many different clinical situations relevant to cancer therapy.

Figure 1. Clearly both groups of GBM patient’s given high

This area of research continues to be a major focus of the

dose vitamin C with radio-chemotherapy show promising

RPOE in Redox Biology and Medicine in the coming year.

results for increasing overall survival, as compared to historical controls. In 2016 these very promising clinical trials combined

The RPOE has also supported projects to develop modern

with the supporting basic science studies (1) have resulted in

magnetic resonance imaging (MRI) techniques to monitor

the submission of a P01 Program project entitled “Exploiting

and predict sensitivity to high dose vitamin C in cancer

Redox Metabolism Using Pharmacological Ascorbate for

patients. The faculty in the RPOE in collaboration with MR

Cancer Therapy” (P01CA217797; Cullen, Spitz MPI;

imaging physicist, Dr. Vince Magnotta, have discovered that

$12,106,162 total costs) and an RO1 application entitled

increased T2* relaxation times from standard MRI scans

“Exploiting Labile Iron Pools for Improving Non-Small Cell

could be used to monitor tissue changes in redox active

Lung Cancer Therapy Using Pharmacological Ascorbate”

iron that would be expected to predict tumor sensitivity to

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high dose vitamin C combined with radio-chemotherapy.

The Phase I clinical trial assessing the tolerability of a ketogenic

This discovery has significantly contributed to both an

diet delivered via PEG tube in combination with concurrent

RO1 (R01CA215732; Allen PI) and PO1 Project 3 (Allen,

radiation and chemotherapy in locally advanced head and neck

project leader) application (P01CA217797; Cullen, Spitz

cancer was only able to complete 4 of 11 subjects enrolled on

MPI). This work will continue to be a focus of the RPOE

the trial. Two of the six subjects who withdrew from the trial

in Redox Biology and Medicine in the next year.

had a DLT from grade IV hyperuricemia (possible diet related). The other four subjects who withdrew did so in the early weeks

In another important translational initiative the RPOE has

of the trial after experiencing nausea, vomiting, fatigue, and/or

been working with an investigator initiated industry supported

general displeasure with the diet or for personal reasons (possibly

study of small molecule superoxide dismutase mimics with the

diet related). These trial results will be published, but the trial

Galera Therapeutics, Inc. (Spitz, PI) for a pre-clinical project

will be discontinued due to the inability to obtain satisfactory

entitled “Enhancing Cancer Cell Killing via H2O2-mediated

dietary compliance to move forward with Phase 2 studies.

Metabolic Oxidative Stress Using SOD Mimics” as well as the completion of a phase I clinical trial entitled “A Phase

A final study supported by the RPOE is focused on

I, Open-Label, Multi-Center, Dose Escalation Study of the

repurposing of currently available relatively non-toxic

Safety, Tolerability, Pharmacodynamic and Pharmacokinetic

FDA approved drugs such as D-penicillamine (DPEN)

Properties of Intravenous GC4419 in Combination with

and disulfiram (DSF), to target fundamental differences

Radiation and Chemotherapy” (NCT01921426; Buatti, PI).

in oxidative metabolism in cancer versus normal cells.

Members of the RPOE at Iowa continue to lead the nation

Preliminary data generated show that DPEN and DSF are

in accrual to this trial but the blinded nature of the study

selectively toxic to breast and lung cancer cells, relative to

precludes data analysis at this time and a recent paper from

normal breast and lung epithelial cells. In addition, DPEN

the basic science work was submitted to Cancer Research

and DSF were found to be capable of enhancing the effects

(4). This work continues to be supported by two industry

of radiation on lung cancer cells. The project lead to a

funded initiatives (for a total of $250,000/year) and will

15th percentile score for an MSTP student F30 application

remain a major focus of investigation in the coming year.

(CA213817) that is expected to be funded in April of 2017.

The RPOE faculties also continue to develop ketogenic diets

Major accomplishments of the members of the Research Program

(KD) as an adjuvant therapy in pancreas, lung, and head and

of Excellence in Redox Biology and Medicine in 2016 include:

neck cancer therapy when combined with radio-chemo-therapy (NCT01419587; NCT01975766; NCT01419483). Currently

• Submission of major grant funding for the high dose

one paper is in the final stages of revision for publication (on

vitamin C translational studies in pancreas, brain,

the pancreas and lung studies) and one is being prepared for

and lung cancer initiative (P01CA217797, Cullen/

submission on the head and neck cancer studies (5). When

Spitz MPI, $12,106,162 total costs; R01CA221292,

these studies were extended to clinical trials with NIH support

Spitz/Allen MPI, $3,356,606 total costs;

(R21CA161182) subjects with locally advanced NSCLC and

R01CA215732, Allen PI, $3,723,673 total costs).

pancreas cancer receiving concurrent radiation and chemotherapy went into ketosis within two days of beginning the diet and during therapy showed increases in some systemic markers of oxidative damage to lipids and proteins. However of the nine subjects enrolled in the Phase I trials only three completed a full course of therapy while consuming the diet demonstrating suboptimal compliance and tolerance for an oral KD. This led the group to extend the study into head and neck cancer (HNSCC) subjects based on preclinical findings hoping to increase compliance since most of these subjects are fed through a PEG tube and it was hoped this would improve compliance.

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• Submission and favorable reviews for a large collaborative paper in Cancer Cell that is very close to acceptance with multiple authors from the RPOE faculty (1) as well as two more collaborative peer-reviewed publications (2, 3). • Continued progress of ongoing basic and clinical studies of SOD mimics supported by Galera Therapeutics as well as submission of a new manuscript in this field to Cancer Research (4).

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

DOUG SPITZ, PHD Balance Forward Revenue Disbursements to UI Total Revenue

($23,648) Other (15.3%)

$100,000

Expenses Personnel $73,085 Lab Supplies/Animals $25,358 Travel $1,414 Equipment $290 Equipment Maintenance $3,900 Other $18,828 Total Expenses $122,876 Ending Balance

Personnel (59.5%)

Equipment Maintenance (3.2%)

Equipment (0.2%)

Travel (1.2%)

Lab Supplies/Animals (20.6%)

$46,524

PUBLICATIONS

Publications relevant to studies supported by the RPOE in Redox Biology and Medicine in 2016:

3. Witmer JR, Wetherell BJ, Wagner BA, Du J, Cullen JJ, Buettner GR: Direct spectrophotometric measurement of supra-physiological levels of ascorbate in plasma. Redox Biol. 2016; 8:298-304. PMID: 26928133

1. Schoenfeld JD, Sibenaller ZA, Mapuskar KA Wagner BA, CramerMorales KL, Furqan M, , Sandhu S, Carlisle TL, Smith MC, Abu Hejleh T, Berg DJ, Zhang J, Keech J, Parekh KR, Bhatia S, Monga V, Bodeker KL, Ahmann L, Vollstedt S, Brown H, Shanahan Kauffman EP, Schall ME, Hohl RJ, Clamon GH, Greenlee JD, Howard MA, Shultz MK, Smith BJ, Riley DP, Domann FE, Cullen JJ, Buettner GR, Buatti JM, Spitz DR*, Allen BA*. O2•- and H2O2-mediated disruption of Fe metabolism causes the differential susceptibility of NSCLC and GBM cancer cells to pharmacological ascorbate. Cancer Cell 2017; (*cocorresponding authors, being considered for publication in the 3nd revision)

2. Doskey CM, Buranasudja V, Wagner BA, Wilkes JG, Du J, Cullen JJ, Buettner GR: Tumor cells have decreased ability to metabolize H2O2: Implications for pharmacological ascorbate in cancer

4. Mapuskar KA, Flippo KH, Schoenfeld JD, Riley DP, Strack S, Abu Hejleh T, Furqan M, Domann FE, Buatti JM, Goswami PC, Spitz DR, and Allen BG: Mitochondrial superoxide mediates age-associated increases in susceptibility of dermal fibroblasts to radiation and chemotherapy. Cancer Res. 2017; (submitted).

5. Zahra A, Opat E, Bhatia SK, Fath MA, Ma DC, Rodman SN, Snyders TP, Eichenberger-Gilmore JM, Chenard CA, Kellie Bodeker KL, Ahmann L, Mott SL, Vollstedt S, Brown H, Abu Hejleh T, Clamon GH, Berg DJ, Spitz DR, Buatti JM, and Allen BG: Consuming a Ketogenic Diet while Receiving Radiation and Chemotherapy for Locally Advanced Lung and Pancreas Cancer: The University of Iowa Experience of Two Phase I Clinical Trials. Radiation Res. 2017; (in revision).

therapy. Redox Biol. 2016; 10:274-284. PMID: 27833040

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MEDICAL RESEARCH INITIATIVE GRANTS

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2017 GRANT AWARDS

CARVER MEDICAL RESEARCH INITIATIVE GRANT AWARDS

STACEY DEJONG, PT, PHD Assistant Professor, Department of Physical Therapy “Operant conditioning to decrease hypertonia and improve wrist motor control after stroke”

SIEGFRIED JANZ, MD, DSC Professor, Department of Pathology “Transgenic mouse model of human KSHV-associated B-lymphocyte disorders”

KEVIN LEGGE, PHD Associate Professor, Department of Pathology “Nanoparticle vaccination against Influenza virus”

ASHUTOSH MANGALAM, PHD Assistant Professor, Department of Pathology “Deciphering the mechanism of obesity induced inflammation in multiple sclerosis utilizing human leucocyte antigen (HLA) class-II transgenic mice”

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JAMES MCNAMARA, PHD Associate Professor, Department of Internal Medicine “Rapid antibiotic resistance profiling of uropathogens”

MARY VAUGHAN SARRAZIN, PHD Associate Professor, Department of Internal Medicine “Dosing of Direct Oral Anticoagulants for stroke prevention in atrial fibrillation”

CARVER COLLABORATIVE PILOT GRANT AWARDS

JEFFREY MEIER, MD

DAVID PRICE, PHD

Associate Professor, Department

Professor, Department of

of Internal Medicine

Biochemistry

“Transcriptional mechanisms controlling cytomegalovirus infection”

RAJAN SAH, MD, PHD

ANDREW NORRIS, MD, PHD

SAMUEL STEPHENS, PHD

Assistant Professor, Department

Associate Professor, Department

Assistant Professor, Department

of Internal Medicine

of Pediatrics

of Internal Medicine

“SWELL1-mediated regulation of pancreatic ß-cell function”

M E D I C A L R E S E A R C H I N I T I AT I V E G R A N T S

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Dr. Bryan Allen’s research focuses on

BRYAN G. ALLEN, MD, PHD

improving responses to cancer therapy, developing strategies for inhibiting and protection of normal tissue injury during cancer therapy and identifying changes in redox metabolism and biomarkers of oxidative stress relevant to predicting

ASSISTANT PROFESSOR, DEPARTMENT OF

RADIATION ONCOLOGY

outcomes following various therapies. The role of reactive oxygen species (ROS) in age-associated susceptibility to radiation (IR) and chemotherapy Funds received from the Roy J. Carver Charitable Trust Pilot Grant in February of 2016 were used to pay salaries, purchase lab supplies, and pay the core lab charges for studies related to evaluating the age-associated changes in redox metabolism that mediate the differences in sensitivity to radiation and chemotherapy. This included: purchasing cell culture media and growth factor kits; antibodies for DNA damage markers; purchase of fluorescence dyes to detect changes in mitochondrial metabolism; and labware including tissue culture flasks, plates, centrifuge tubes, flow cytometry tubes, etc. A part of the expenses were also used to cover the usage of the university core services including: flow cytometry usage charges; radiation facility usage (facility used for all experiments involving the use of low-dose radiation); and radiation and free radical research core usage (facility used to test samples for antioxidant enzyme activity levels) charges. The main goal of this grant was to gather preliminary data for the hypothesis that age-associated increased levels of reactive oxygen species (ROS), more specifically superoxide mediate the increased susceptibility of fibroblasts from old donors to IR and chemotherapy that can be mitigated by a SOD mimetic. The results obtained so far demonstrate that 1. Fibroblasts from elderly (old) male donor subjects (70, 72, 78 years) are significantly more sensitive to killing mediated by platinum-based chemotherapy and IR, relative to young fibroblasts (5 months and 1 year) and/or adult fibroblasts (20 year old).

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February 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

BRYAN G. ALLEN, MD, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$0

Lab Supplies (25.7%)

Travel (0.3%) Other (3.6%)

$30,000

Personnel (70.4%)

Expenses Personnel $12,548 Lab Supplies $4,588 Travel $56 Other $640 Total Expenses $17,832 Ending Balance

$12,168

2. The data reveals that fibroblasts from old donors

These results support Dr. Allen’s hypothesis that, age-

show changes in mitochondrial metabolism

associated increased superoxide and resulting DNA damage

including increased steady-state levels of ROS, more

mediate the increased susceptibility of old fibroblasts to IR

specifically superoxide (a form of ROS), oxygen

and chemotherapy that can be mitigated by GC4419.

consumption and ATP levels as well as decreased mitochondrial electron chain complex (ETC)

The above results were presented at a poster

and aconitase (Tricarboxylic Acid cycle enzyme)

presentation at the Radiation Research Society

activities and altered mitochondrial structure.

(RRS) conference in October 2016.

3. Mitochondrial ETC activity and aconitase activity

The poster can be found here:

were restored by manganese superoxide dismutase

https://drive.google.com/

(SOD2)—(an antioxidant known to scavenge superoxide)

open?id=0B8XBb5fkTQWMbm41d2g4bFppMms

overexpression clearly demonstrating a role for mitochondrial superoxide in these effects. Fibroblasts

MANUSCRIPT SUBMITTED TO CANCER

from old donors also demonstrated elevated levels

RESEARCH:

of basal DNA damage that were further increased

Kranti A. Mapuskar, Kyle H. Flippo, Joshua D.

following treatment with IR and chemotherapy.

Schoenfeld, Dennis P. Riley, Stefan Strack, Taher Abu Hejleh, Muhammad Furqan, Frederick E. Domann, John

4. Most importantly, treatment with a small-molecule, superoxide-specific (superoxide dismutase) SOD mimetic (GC4419) mitigated the increased sensitivity of old fibroblasts to IR and chemotherapy as well as partially

M. Buatti, Prabhat C. Goswami, Douglas R. Spitz*, and Bryan G. Allen* Mitochondrial superoxide mediates age-associated increases in susceptibility of dermal fibroblasts to radiation and chemotherapy.

restored mitochondrial function without affecting IR or chemotherapy-induced cancer cell killing.

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BOTOND BANFI, MD, PHD ASSOCIATE PROFESSOR, DEPARTMENT OF ANATOMY

AND CELL BIOLOGY

ASSOCIATE PROFESSOR, DEPARTMENT OF INTERNAL MEDICINE ASSOCIATE PROFESSOR, DEPARTMENT OF

OTOLARYNGOLOGY— HEAD AND NECK SURGERY

Dr. Botond Banfi’s research focuses on the identification and functional characterization of genes that are critical for hearing. The goal of this research is to gain new insights into the molecular mechanism of hearing and to find novel drug targets for the prevention of hearing loss.

A novel deafness gene encodes a drug-responsive regulator of transcription Funds from the Roy J. Carver Charitable Trust were used in the Banfi lab to accomplish two goals: 1) test whether a gene expression-modifying drug can be used for the prevention of hearing loss in mice that harbor the same deafness-causing mutation as a group of deaf human subjects; and 2) establish a method for the isolation of pure populations of sensory cells from the mouse inner ear. Carver Trust funds were spent in course of this study to: 1) pay the cost of housing of mutant and control mice; 2) pay the cost of reagents; and 3) pay part of the salary of a research scientist who conducted the experiments.

Hereditary hearing loss is one of the most common birth defects, affecting about 1 in 1,000 babies worldwide. The identification of deafness-causing mutations and the characterization of affected genes has been a fruitful approach for gaining insights into the molecular mechanisms underlying the development of sensory cells in the ear, and a key step towards developing new therapies. However, the pathogenic mutations are typically in genes whose products cannot be manipulated favorably with existing drugs. Preliminary data from the Banfi lab suggested that a hearing loss-causing mutation in the human REST gene could be an exception. The human REST mutation was discovered as a result of a collaborative effort in the Banfi (UI) and Friedman (NIH/

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NIDCD) labs. The affected protein REST is known to regulate the production of hundreds of proteins that are normally present only in neuronal cells. In nonneuronal cells, the level of REST protein is high. REST binds to the genes of neuronal proteins and inhibits their expression. In developing neurons, the neuronal proteins are upregulated (“derepressed”) because REST is down-regulated. The hearing loss-causing mutation in the human REST gene impairs one of the mechanisms that down-regulates REST, suggesting that REST is too active in cells that rely on the affected mechanism for the downregulation of REST. REST inhibits mRNA production from its target genes in a complex with a group of enzymes called histone deacetylases (HDAC). Several HDAC inhibitors have been developed in recent years. The central hypothesis of this study was that HDAC inhibitors can be used to prevent hearing loss in a genetically modified mouse line that harbors the same defect in the REST gene as a group of deaf human subjects. In addition, Dr. Banfi wished to establish a method for the collection of pure sensory-cell populations from the mouse inner ear. This was an important goal because mRNA expression studies in pure cell populations may lead to the identification of REST target genes that are critical for hearing. The proposed project had two aims.

Aim 1 Determine the extent to which HDAC inhibitors affect hearing loss and sensory cell degeneration in a mouse model of the human REST defect. SAHA is the only known HDAC inhibitor that permeates the inner ear after systemic administration. Therefore, SAHA was used in the experiments with REST mutant mice. Since the sensory cells degenerate in the REST mutant mice perinatally, the SAHA treatment was started on postnatal day (P)1 Figure 1. On P5 the morphology of sensory cells was evaluated in the inner ear of SAHA-treated REST mutant mice, non-treated REST mutant mice, and non-treated control mice (wild-type). The sensory cells were visualized with an actin-binding fluorescent dye. This experiment revealed that the majority of sensory cells had normal or nearly-normal morphology in the SAHAtreated REST mutant mice. In contrast, the sensory cells of non-treated REST mutant mice degenerated Figure 1. Thus, short-term SAHA treatment was effective in preventing the degeneration of sensory cells of REST mutant mice.

M E D I C A L R E S E A R C H I N I T I AT I V E G R A N T S

Figure 1. SAHA rescues the sensory cells from degeneration in REST mutant mice. Phalloidin-Alexa488-based visualization of actin reach stereocilia bundles and cell-cell borders in the organ of Corti from a nontreated wildtype (WT) mouse, a nontreated REST mutant mouse, and a SAHAtreated REST mutant mouse at P5. SAHA (100 mg/kg body weight) was injected subcutaneously three times after birth, as illustrated by the scheme on the top. The “non-treated” mice received only the solvent of SAHA (i.e, a mixture of DMSO, PBS, and Cremophor EL). In the nontreated REST mutant mouse the sensory cells lost their stereocilia bundles and are abnormally swollen, indicative of cell degeneration.

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In wild-type mice, the onset of hearing occurs on P12-13. To evaluate the effect of SAHA on the hearing of REST mutant mice, various SAHA administration regimens were designed for the duration of the first two postnatal weeks. The hearing of SAHA-treated mice and non-treated mice was measured at P16, and the hearing thresholds were compared with that of non-treated wild-type mice. The hearing threshold of nontreated REST mutant mice was more than 50 decibel above normal (i.e., wild-type) levels, indicating complete hearing loss. The most effective SAHA regimen, which included dose escalation after P12, reduced the hearing threshold of REST mutant mice to 22Âą4.3 decibel above normal levels. Thus, the SAHA treatment rescued the hearing of REST mutant mice. Aim 2 Determine the effect of the REST mutation on gene expression in the sensory cells of the inner ear. During 2016, the Banfi lab established a method for the isolation of mouse sensory cells. The method is based on the fluorescent dye (FM1-43)-labeling of sensory cells of P3 mice, the enzymatic dissociation of the organ of Corti, and the sorting of fluorescent versus non-fluorescent cells. In the first half of this year (2017), this experiment is planned to be scaled up to the level that sufficient number of sensory cell can be isolated for transcriptome-wide analysis of gene expression. The above described data were generated for the revision of an NIH R01 grant application that was funded in 2016 (1R01DC014953, PI: Banfi). In addition, these data (along with other results) will be submitted for publication in the first quarter of 2017.

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February 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

BOTOND BANFI, MD, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$0

Other (1%) Personnel (61.6%)

$30,000

Expenses Personnel $18,482 Lab Supplies/Animals $11,216 Other $302 Total Expenses $30,000 Ending Balance

$0

M E D I C A L R E S E A R C H I N I T I AT I V E G R A N T S

Lab Supplies/Animals (37.4%)

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JOHN D. COLGAN, PHD Dr. John Colgan’s research focuses on ASSOCIATE PROFESSOR, DEPARTMENT OF INTERNAL

understanding pathways that control

MEDICINE

immune cell development and function. His

ASSOCIATE PROFESSOR, DEPARTMENT OF ANATOMY

work will help find ways to boost protection

provided by the immune system and to

AND CELL BIOLOGY

treat diseases that arise when the immune system functions improperly or gives rise to cancerous cells. Control of peripheral T cell responses by the developmental regulator GON4L Funds from the Carver Trust were used to explore the importance of the gene regulator GON4L in T cells, which are critical components of the immune system. Thus far, Dr. Colgan’s work has demonstrated GON4L is needed during the early stages of T cell development and for the formation of mature T cells that mediate immune defense. GON4L is a nuclear protein containing domains found in factors that alter chromatin structure, suggesting a role in gene regulation Fig. 1. In keeping with this idea, Dr. Colgan’s work showed GON4L associates with factors that regulate chromatin and gene expression. Genetic studies of plants, worms, and zebrafish and Dr. Colgan’s work on mouse B cell development all suggested GON4L regulates cell fate decisions and cell division in the context of developmental pathways. Supporting a role in cell division, GON4L expression is elevated in a variety of human cancers and drives the growth of cancer cell lines in culture. These findings suggest GON4L has a dual role and regulates genes essential for cell differentiation and division. Yet, the function of GON4L and the pathways that require it are not understood. Dr. Colgan is addressing this knowledge gap using unique mouse models to probe GON4L function during T cell development and mature T cell responses.

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Fig. 1. The mouse GON4L protein. The N-terminus has a nuclear localization signal (NLS) and homology to histone chaperones. Two C-terminal regions match the paired amphipathic helix (PAH) repeat consensus. The transcriptional co-repressors SIN3A and -3B contain PAH repeats that interact with chromatin-modifying factors or DNA-binding proteins. The C-terminus forms a SANT domain, which is found in several transcriptional regulators. Fig. 2. Loss of GON4L in primitive precursors blocks T cell development. All data were obtained from thymii harvested from control or Lck-cre Gon4lD/D (KO) mice. (A) Picture of freshly isolated thymii. (B) Cell yields. (C, D) Flow cytometric analysis of late (C) and early (D) T cell development. Panels at the top show the developmental path of cells take. (E) Yields of DN4-stage T cell precursors. Data are representative of results from 5 independent experiments. **, P < .01; ***, P< .001.

Results Dr. Colgan’s studies thus far have focused on the importance of GON4L for the development of CD4 and CD8 T cells. Two novel strains of mice were created for this work. The first, known as Lck-cre Gon4lD/D mice, inactivate the gene encoding GON4L during the earliest stages of T cell development as they occur in the thymus. The second strain, called CD4-cre Gon4lD/D mice, inactivate the GON4L gene during the final stages of T cell development in the thymus and in mature CD4 and CD8 T cells that populate the spleen and lymph nodes and support immune defense. Results obtained from each of these mouse models are described below. Lck-cre Gon4lD/D mice: T cell development within these mice was analyzed by characterizing the thymus, the site of where T cells are first generated. Thymii from Lck-

controls Fig. 2A and contained much fewer cells Fig. 2B. Flow cytometric analysis showed that cells at the DP, CD4 SP and CD8 SP stages of T cell development were greatly reduced in Lck-cre Gon4lD/D mice Fig. 2C, indicating T cell development is blocked by the loss of GON4L. The DN subset contains primitive T cell precursors, so this fraction was further analyzed. DN cells can subdivided into DN1, DN2, DN3 and DN4 fractions, with DN1 cells being the most primitive Fig. 2D. This showed the loss of GON4L did not affect DN1, -2 or -3 cells, but greatly impaired the development of DN4 cells, as manifested by greatly reduced numbers of DN4 cells in Lckcre Gon4lD/D mice compared to controls Fig. 2E. The DN3 to DN4 transition is a crucial stage; here cells begin expressing T cell-specific receptors required for further development. Thus, Dr. Colgan’s work has demonstrated GON4L is essential for a critical step in the early stages of T cell development.

cre Gon4lD/D mice were small compared to those from

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Fig. 3. Loss of GON4Limpairs formation of mature T cells. All data were obtained from spleens harvested from control or CD4-cre Gon4lD/D (KO) mice. (A) Flow cytometric analysis of cell subsets in spleen. (B). Yields of the indicated cell populations from spleens. (C) Flow cytometric analysis of CD4 and CD8 T cell in spleen. (D) Yields of CD4 and CD8 T cells from spleen.

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February 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

JOHN D. COLGAN, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$0

Equipment (7.9%)

$30,000

Expenses Personnel $11,928 Lab Supplies $895 Equipment $1,093 Total Expenses

$13,915

Ending Balance

$16,085

Lab Supplies (6.4%)

Personnel (85.7%)

CD4-cre Gon4lD/D mice: In these mice, GON4L is lost late in T cell development and should be absent in mature CD4 and CD8 T cells. Therefore, the late stages of T cell

PENDING GRANT

development in thymus and mature CD4 and CD8 T cells in spleen were examined using flow cytometry. Only subtle differences in cells from thymus were noted, arguing GON4L is not essential for the final stages of T cell development (data not shown). In spleen, the numbers of non-T cells (i.e. B cells and myeloid cells) were normal in CD4-cre Gon4lD/D mice, but the numbers of T cells were significantly reduced Fig 3A & 3B, indicating formation of mature T cells was disrupted by

NIH R21 AI125817 Colgan (PI) Control of peripheral T cell responses by the developmental regulator GON4-like The goals of this project are to: 1) determine how GON4L directs the formation and function of peripheral CD4 T cells; 2) determine how GON4L directs the formation and function of peripheral CD8 T cells.

the loss of GON4L. Mature T cells consist of both CD4 and CD8 subsets, which are generated by different developmental pathways and have distinct functions. The numbers of CD4 and CD8 T cells were significantly reduced in spleens from CD4-cre Gon4lD/D mice Fig 3C & 3D, which indicates that loss of GON4L impairs the generation of both types of T cells. From this work, Dr. Colgan has shown that GON4L has an important role in the formation of mature CD4 and CD8 T cells that provide immune defense. These findings provide justification for continuing with the project by analyzing how mature CD4 and CD8 T cell responses to infections are affected by the loss of GON4L.

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FANG LIN, MD, PHD Dr. Fang Lin’s research focuses on understanding the role of a gene called ASSOCIATE PROFESSOR, DEPARTMENT OF

ANATOMY & CELL BIOLOGY

brain-specific angiogenesis inhibitor 2 (bai2), a G protein-coupled receptor in heart development.

Novel roles of brain-specific angiogenesis inhibitor 2 in left-right asymmetry Funds from the Roy J. Carver Charitable Trust were used in 2016 to generate genetic mutant lines and to characterize the phenotypes of these mutants. The 2016 Carver Trust funds were used to elucidate the role of a protein family known as brain angiogenesis inhibitor (bai) in the cardiovascular system. Previous data found that one member of the bai family, bai2, is expressed in cells that influence the direction in which the heart grows, allowing the heart to develop normally. Asymmetrical formation of the heart and other organs is essential for proper development and, when it is disrupted, can lead to lethal defects in the organism. To understand the role of bai2 protein in heart development, Dr. Lin’s laboratory generated zebrafish animals in which different areas in the gene were mutated so that the gene does not produce functional proteins. The founders and the animals of the first generation exhibited similar deviations in heart placement—leaning to the left, right, or remaining in the center at random, implicating that bai2 plays a role in establishing normal heart positioning. However, phenotypes were lost in future mutant generations, suggesting that there are compensatory mechanisms that allow the heart to develop normally. One possibility is that the second isoform of the gene plays a redundant role. To test this possibility, another mutant line harboring the non-functional second gene was acquired and crossed into the first mutant animal. An animal line in which both genes are mutated are being generated and their heart development will be assessed.

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February 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

FANG LIN, MD, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$0

30,000

Expenses Personnel $7,574 Lab Supplies/Animals $16,878 Total Expenses $24,452 Ending Balance

Lab Suppplies/ Animals (69%)

Personnel (31%)

$5,548

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Dr. Christie Thomas’ research focuses on the vascular endothelial growth factor (VEGF) and its receptors FLT1 and KDR. Dr. Thomas is examining the function of the receptor FLT1 and the ways in which it may

CHRISTIE P.

regulate VEGF function. His work is relevant

THOMAS, MBBS

preeclampsia.

for vascular biology, hypertension, and

MicroRNA regulation of SFLT1 expression in the PROFESSOR, DEPARTMENT OF INTERNAL MEDICINE

placenta Funds from the Roy J. Carver Charitable Trust were used in 2016 to further study the cleavage of the receptor FLT1 and Dr. Thomas was able to show that this cleavage regulates activation of the receptor and its function. He is still working on how the ligand VEGF-A inhibits this cleavage.

Flt1 is a cell surface VEGF receptor cleaved to release an N-terminal ectodomain, which binds VEGF and PlGF and can antagonize the effects of VEGF in the extracellular milieu. To further evaluate Flt1 processing we expressed tagged Flt1 constructs in HEK293 and COS7 cells where we demonstrate, by deletion mapping, that the cleavage site is immediately adjacent to the transmembrane domain (TMD) between residues 759 and 763. Cleavage reciprocally regulates free VEGF in conditioned media and we show that the cleavage site is also transferable to another transmembrane receptor. A second cleavage event downstream of the ectodomain cleavage releases a cytosolic C-terminal Flt1 fragment and this intracellular cleavage of Flt1 is not catalyzed or regulated by the upstream ectodomain cleavage since abolition of the ectodomain cleavage has no impact on the downstream cleavage event. The downstream cleavage event is not susceptible to γ-secretase inhibitors and overexpression of presenilin 1, the catalytic subunit of γ-secretase did not change the downstream intracellular cleavage event. Furthermore, this cleavage did not occur via a previously published valine residue (767V) in the TMD of Flt1, indicating the existence of another cleavage pathway. We tested the impact of the ectodomain cleavage on p44/42 MAP kinase activation and demonstrate that compared to wild type Flt1, cleavage resistant Flt1 constructs failed to stimulate

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February 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

CHRISTIE THOMAS, MBBS Balance Forward Revenue Disbursements to UI Total Revenue

$0

Other (4%)

$30,000 Lab Supplies (96%)

Expenses Lab Supplies $10,641 Other $442 Total Expenses $11,083 Ending Balance

$18,917

p44/42 MAP kinase activation. Our results indicated that

of the VEGF inhibitory effect. Our results indicate that

Flt1 ectodomain cleavage not only regulates the availability

the inhibition of FLT1 ectodomain cleavage by VEGF-A is

of free VEGF in the extracellular milieu but also regulates

neither dependent on receptor activation, or internalization

downstream signaling via the ERK kinase pathway.

nor a consequence of receptor degradation and likely represents a direct inhibitory effect on receptor cleavage

We then evaluated the effect of VEGF-A on FLT1 cleavage

mediated by ligand stimulated receptor dimerization.

with tagged and untagged constructs in native and transient and stable expression systems. We demonstrate that

The Carver funds have helped considerably this past year.

VEGF-A inhibits FLT1 ectodomain cleavage in a time- and

The second of two papers is in review and they are working

dose-dependent manner while VEGF-A knockdown in

to complete some experiments in response to the reviewers’

HEK293 cells increases ectodomain shedding. Although

comments.

KDR, analogous to FLT1, is also subject to extracellular and intracellular cleavage, VEGF-A does not inhibit KDR cleavage. VEGF-A inhibition of FLT1 cleavage is

PUBLICATIONS

not dependent on FLT1 tyrosine kinase activity or the intracellular FLT1 residues. ALLN, a proteasomal inhibitor,

1. Raikwar, N.S., Liu, K.Z. and Thomas, C.P. Ectodomain cleavage of

bafilomycin A, an inhibitor of endosomal acidification and

FLT1 regulates receptor activation and function and is not required

dynasore, an dynamin inhibitor all increase the abundance

for its downstream intracellular cleavage. Experimental Cell Research 2016

of FLT1 and the shed ectodomain indicating that FLT1 is

pii: S0014-4827(16)30060-X. doi: 10.1016/j.yexcr.2016.03.020.

subject to dynamin-mediated endocytosis and susceptible to proteasomal and lysosomal degradation. VEGF-A inhibition of cleavage is not reversed by ALLN, bafilomycin A or dynasore. However, a 30 AA deletion in the extracellular immunoglobulin 7 domain that appears to inhibit receptor

2. Raikwar, N.S., Liu, K.Z., Shibuya, M. and Thomas, C.P. VEGF-A selectively inhibits FLT1 ectodomain shedding independent of receptor activation and receptor endocytosis 2017 (in review, PLoS One)

dimerization leads to enhanced cleavage of Flt1 with loss

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MEI-LING

NANDAKUMAR

JOINER, MSc, PHD

NARAYANAN, MD, PHD

RESEARCH ASSISTANT PROFESSOR, DEPARTMENT

ASSISTANT DIRECTOR, RESIDENCY PROGRAM IN

OF MOLECULAR PHYSIOLOGY AND BIOPHYSICS

NEUROLOGY ASSISTANT DIRECTOR, CLINICAL NEUROSCIENTIST

TRAINING PROGRAM IN NEUROLOGY ASSISTANT PROFESSOR, DEPARTMENT OF NEUROLOGY

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Parkinson’s disease is a neurodegenerative disorder characterized by a progressive and selective loss of dopaminergic neurons in the substantia nigra pars compacta (SNc). Drs. Mei-Ling Joiner and Nandakumar Narayanan’s research focuses on preserving mitochondrial function in the neurons that are affected by Parkinson’s disease with the aim to reduce the age-related loss of dopaminergic neurons. reduce cell death, and with some treatments may even promote Inhibition of mitochondrial calcium uptake to

cell death. The expression of dominant-negative MCU

reduce development of PD

may generate persistent excessive levels of cytosolic Ca2+,

Funds from the Roy J. Carver Charitable Trust were used to accomplish baseline studies establishing growth conditions of a dopaminergic cell line and level of treatment of these cells to mimic conditions of Parkinson’s disease. The effects of this treatment were measured using various assays for viability and whole cell and mitochondrial function. General inhibitors of mitochondrial Ca2+ uptake were applied to show whether increased mitochondrial Ca2+ promotes decreased cell function or cell death. One of the roles of mitochondria is to maintain appropriate concentrations of Ca2+ within the cell. The mitochondrial calcium uniporter (MCU) allows Ca2+ into the mitochondria matrix. Under stress conditions, more cytosolic Ca2+ is present in the cell, which can cause excessive Ca2+ uptake by the MCU. It is thought that this excess mitochondrial Ca2+ uptake may promote Parkinson’s disease. A mechanism that regulates mitochondrial Ca2+ uptake in dopaminergic cells would reveal a new target for therapeutic strategies to halt the progressive degeneration of SNc neurons in Parkinson’s disease. One such mechanism would be to inhibit MCU and test if it protects from high levels of cytosolic Ca2+. They used SH-SY5Y cells, a dopaminergic cell line, treated with the dopaminergic neurotoxin 6-hydroxydopamine. For this experiment, they inhibited the activity of the MCU by expressing a dominant-negative gene variant, acutely preventing Ca2+ uptake into the mitochondria matrix. They

generating stress conditions that ultimately lead to cell death. They are using both a plate reader based assay and confocal fluorescent microscopy to study cytosolic and mitochondrial Ca2+ level changes following 6-hydroxydopamine treatment. Initial results show that 6-hydroxydopamine causes an increase in both basal cytosolic and basal mitochondrial Ca2+ levels. To show whether mitochondrial function is changed by 6-hydroxydopamine treatment we measured the oxygen consumption rate of treated and control dopaminergic cells. The oxygen consumption rate is a measure of mitochondrial bioenergics or energy production by the cell. 6-hydroxydopamine treatment reduced both the basal level of energy production and spare respiratory capacity compared to that of control cells. The spare capacity is thought to be a parameter necessary for cells to survive stressors such as Ca2+ overload and reactive oxygen species. Interestingly, pretreatment with the MCU antagonist, ru360, increased spare respiratory capacity of the dopaminergic cells. With the help of Ms. Stephanie Alberico in Dr. Kumar Narayanan’s lab they are learning how to stereotactically inject 6-hydroxydopamine into the SNc of the mouse brain in order to expand our in vitro studies to the mouse in vivo model. Transgenic mice engineered to take up Ca2+ into the mitochondria of SNc neurons at a reduced rate are now ready for these injections. This study will test the effect of mitochondrial Ca2+ entry in SNc neurons.

found that the expression of dominant-negative MCU does not

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Studies looking at the effects of blocking MCU function on cell viability generated by SROP student, Ms. Nicole de la Rosa Gonzalez, were presented at a number of symposia, including the 2016 Annual Biomedical Research Conference for Minority Students and the 2016 Symposium for Honor Studies Programs of Puerto Rico. She is also planning to present at the 2017 Experimental Biology meeting. Nicole will continue her studies on this research; she plans to expand the types of assays used to include measurements of mitochondrial calcium, membrane potential, and reactive oxygen species generation. The goal of her project is to understand the molecular pathway leading to cell death in a cellular model of Parkinson’s disease. Through her continued research efforts in my lab, she should be able to contribute to a peer-reviewed article, which will greatly enhance her career development. Her experience in the lab will enhance her application to the University of Iowa Neuroscience graduate program. Among the key components of the Carver College of Medicine 2014–2017 strategic diversity road map is to attract, recruit, retain and successfully graduate a diverse body of students, trainees, residents, and fellows. Nicole is underrepresented as a female resident of Puerto Rico. Her summer internships on this project will address the human capital component of the strategic plan for increasing diversity at the University of Iowa.

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February 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

MEI-LING JOINER, MSc, PHD & NANDAKUMAR NARAYANAN, MD, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$0

Other (10.6%)

$50,000 Lab Supplies (89.4%)

Expenses Lab Supplies $4,145 Other $490 Total Expenses $4,635 Ending Balance

$45,365

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JOSEPH W.

MADHAVAN

TUREK, MD, PHD

RAGHAVAN, PHD

ASSISTANT PROFESSOR, DEPARTMENT OF SURGERY—

PROFESSOR, DEPARTMENT OF BIOMEDICAL

ENGINEERING

CARDIOTHORACIC SURGERY

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February 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

JOSEPH W. TUREK, MD, PHD & MADHAVAN RAGHAVAN, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$0

Other (1.1%)

Personnel (9%)

$50,000 Lab Supplies Animals (83.8%)

Expenses Personnel $2,393 Lab Supplies/Animals $22,223 Travel $1,591 Other $297 Total Expenses $26,504 Ending Balance

Travel (6%)

Lab Supplies (17.6%)

$23,496

The research of Drs. Joseph Turek and

studies were undertaken to assess flow monitoring against the

Madhavan Raghavan focuses on enhancing

current standard of care (pressure monitoring) in a higher-

safety in the use of ECMO, a life-saving

fidelity simulation system. The higher-fidelity system utilizes human blood as the medium and obstructs the oxygenator

device that supports the heart and lungs.

using preformed microspheres, creating sequential levels of

His studies look at a novel flow monitoring

obstruction in the device. Preliminary data obtained from the

technique to warn of catastrophic events in the device before they happen.Â

study period under funding revealed similar correlation of our flow monitoring technique with that of intermittent pressure measurements. If shown to be an effective marker in vivo, implementing flow monitoring in the ECMO clinical setting is feasible and straightforward, and has the potential to save lives.

Novel method for ECLS oxygenator monitoring Funds from the Roy J. Carver Charitable Trust were used to

Funds were primarily used to purchase supplies for the

complete the low-fidelity ECMO simulation study that revealed

simulation ECMO device. The purchase of oxygenators

changes in flow (our proposed measurement) were as good as

accounted for $20,000 of the funds. Additional funds

changes in pressure (the industry standard) to detect device

paid for circuit tubing and microspheres for the

failure. Additionally, the flow changes are non-invasive and

studies. Travel to the national ECMO meeting and to

continuous, drawbacks to the current system of intermittent

present this work was also paid for by this grant.

pressure monitoring. The project was submitted for an American Heart Association The experimental studies in a low-fidelity system demonstrated

Collaborative Sciences Award, a highly competitive 3-year,

that monitoring the flow in the ECMO device has the potential

$750,000 award. It has been selected as a finalist for the

to serve as an effective indicator of ECMO device failure.

award after making it through the letter of intent phase,

Flow monitoring is safe, hands-free, and continuous. Further

with final decision on funding to be made in April 2017.

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DENICE

LEONID V.

HODGSON-ZINGMAN, MD

ZINGMAN, MD

ASSOCIATE PROFESSOR, DEPARTMENT OF INTERNAL MEDICINE—CARDIOVASCULAR MEDICINE ASSOCIATE PROFESSOR, DEPARTMENT OF

ASSOCIATE PROFESSOR, DEPARTMENT OF INTERNAL MEDICINE—CARDIOVASCULAR MEDICINE

BIOMEDICAL ENGINEERING

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This project from the Hodgson-Zingman and Zingman laboratories focuses on understanding how the acceleration of heart rate that occurs with exercise works to improve heart function, particularly in the setting of heart failure. In mice and humans, the project will study heart rate delivered through a pacemaker in order to identify molecular responses and clinical outcomes that will inform whether the strategy could be used as a therapy for heart failure. Episodic Accelerated Pacing of the Atria (EAPA) to induce favorable remodeling in heart failure Funds from the Roy J. Carver Charitable Trust award were used to support the salary of a research assistant who performed and analyzed animal experiments; the purchase and care of laboratory animals; and for laboratory supplies needed to conduct the animal experiments. Funds were used for the human portion of the project to reimburse travel of our patient volunteers and to cover the costs of clinical tests needed to assess efficacy of the pacing strategy but not covered by the volunteers’ insurance. Both the animal and human experiments have already produced significant preliminary results used to apply for an NIH grant on the same, but expanded, project and to support a patent application for the idea.

Figure 1. EAPA in mice. A) Representative intrinsic (NSR) and paced surface ECGs, p: atrial electrogram, qrst: ventricular electrogram, AP: atrial pacing stimulus artefact. B) Schematic of pacing protocol. HR: heart rate.

The following paragraph is a summary of the findings thus far under the Carver Trust award as they were presented in support of an application to the National Institutes of Health for continued funding of this project under the R01 mechanism. This application was submitted in December 2016 and will be reviewed by the NIH in March 2017. These findings were also submitted to the US Patent Office in

exercise activity is sufficient for cardiovascular conditioning. However, our preliminary data indicate that the typical exercise pattern and extent of HR acceleration can also be a driver of mechanisms promoting myocardial stress resistance

support of a patent application for the pacing technology.

and metabolic efficiency. As such, the central hypothesis is

Heart disease and heart failure (HF) are major public health

exercise, but also an independent physiologic signal that can

challenges. Exercise is a powerful way to prevent heart disease

be harnessed to induce beneficial myocardial adaptations.

and is increasingly understood to mitigate adverse outcomes in

Understanding of the role of episodic HR acceleration as a

HF. The magnitude and duration of exercise heart rate (HR)

physiologic signal could lead to new, titratable, cardiac-specific,

increase are commonly viewed as indicative of whether an

beneficial interventions to complement established therapies

M E D I C A L R E S E A R C H I N I T I AT I V E G R A N T S

that episodic HR acceleration is not just a marker for effective

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for heart disease. Specifically, this proposal studies episodic accelerated pacing of the atrium (EAPA) in both animals and human subjects. An integrated approach is pursued to provide the greatest power to define mechanisms (murine genetic models) while establishing the translational potential in the most direct and clinically meaningful manner (human subjects with chronic HF). Preliminary results in mice indicate that EAPA Fig. 1 increases expression of cardioprotective ATPsensitive potassium (KATP) channels coupled with inhibition of calcium/calmodulin-dependent protein kinase II (CaMKII). Figure 2. IR injury. Representative TTC stains for viability and summary statistics of infarct size as % of total area of sham vs. EAPA treated mice (n=4 each) *p<.05.

EAPA also upregulates peroxisome proliferator-activated receptor γ coactivator 1α (Pgc1α) and optic atrophy 1 (OPA1), known regulators of mitochondrial remodeling and apoptosis resistance. These changes are associated with increased cardiac energetic efficiency and resistance to ischemia-reperfusion (IR) injury Fig. 2. In mice with heart failure due to myocardial infarction with ventricular dysfunction, EAPA demonstrates trends toward reduced fibrosis and improved ejection fraction Fig. 3. EAPA delivered in six human subjects with chronic, medically refractory HF, using the pacing function of their implanted defibrillators Fig. 4, is well-tolerated Table 1. Measures of functional capacity and quality of life improved in two such subjects that underwent four weeks of EAPA sessions. Initial evidence of benefit in healthy and diseased murine hearts, tolerability and trends toward improved outcomes in human subjects with severe HF, and the relatively low risk and cost of atrial pacing in those who do not already have such devices, all suggest potentially rapid, broad, low-cost, thus high impact, applicability of EAPA as a therapeutic intervention and scientific tool if further validation is successful. Significant progress on the basic science and clinical arms of the research project has been made in 2016. The findings supported by the Carver Trust award form the basis for an application to the NIH for a 5-year major research project award (R01) that will be reviewed in March 2017

Figure 3. Murine infarct model. A, B) Representative mid-chamber infarcted mouse heart sections with Masson Trichrome stain. C) Fibrotic area of left ventricles (LV, n=3 each) calculated from stacked sections as represented in A,B) using Image J software. D) LV ejection fraction by echocardiography (n=2 to 4 each, see text). *p<.05.

and also an application to the US Patent Office to secure intellectual property for the University of Iowa for the EAPA invention. Dr. Hodgson-Zingman and Dr. Zingman plan to continue to obtain animal and clinical data with the remainder of the Carver Trust award in order to pursue this exciting line of investigation. Short-term goals are to secure extramural funding for the project and to publish the results in the coming year. Long-term goals are to translate the findings to clinical use, if indicated, through cooperation with pacemaker manufacturers on development of automated protocols and a larger clinical trial.

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February 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

DENICE HODGSON-ZINGMAN, MD & LEONID V. ZINGMAN, MD Balance Forward Revenue Disbursements to UI Total Revenue

$0

Other (4.5%)

$50,000 Personnel (71.1%)

Expenses Personnel $22,904 Lab Supplies/Animals $6,747 Travel $1,119 Other $1,434 Total Expenses $32,204 Ending Balance

Travel (3.5%)

Lab Supplies/ Animals (20.9%)

$17,796

Figure 4. Clinical trial. Schematic of EAPA vs. Sham pacing by manual reprogramming of implanted pacing devices and collection of hemodynamic data in human subjects. Subjects underwent the protocol 3-5 d/ wk for 4 wks. Representative bioimpedance tracings of the HR, CO and TPR response during a single episode of EAPA are shown on the right. HR: heart rate, CO: cardiac output, TPR: total peripheral resistance.

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YOUNG INVESTIGATOR AWARDS

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HANK QI MD, PHD Dr. Hank Qi’s research focuses on dissecting ASSISTANT PROFESSOR, DEPARTMENT OF ANATOMY

AND CELL BIOLOGY

the role of the epigenetic regulations in the development and drug resistance of breast cancer and prostate cancer. Epigenetic modifications dynamically affect differential expression of genes, of which, the protein products are important for both development and drug resistance of cancer cells. Dr. Qi aims to discover the epigenetic factors as novel therapeutic targets for breast and prostate cancers. The Epigenetic Role of Histone Demethylase PHF8 in the Signaling Pathways Associated with Autism, Cleft Lip/Palate, and Prostate Cancers Funds from the Roy J. Carver Charitable Trust were used to complete the last phase of the proposed studies: the epigenetic mechanisms in cancer development. The researches in both breast and prostate cancer studies have progressed successfully toward publications. In detail, the funds from the Carver Trust were used to support a postdoctoral fellow (Peng Shao) and a PhD student (Peterson Kariuki Maina) for breast cancer and prostate cancer research, respectively. The funds were used for their salaries, reagents, and mouse works. The reagents included essential chemicals for cell culture, biochemical approaches, and core facility use such as DNA sequencing, oligonucleotide synthesis, flow cytometry analysis and microscopic analysis. In general, the

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funds of Carver Trust greatly supported the research activities

research direction for federal funding application in 2017.

in Dr. Qi’s Lab and lead to four major accomplishments: Last, Dr. Qi and his PhD student Peterson Kariuki First, in the breast cancer study, Dr. Qi and postdoctoral

Maina responded a collaboration request and

fellow Dr. Peng Shao made great progress in dissecting the

contributed to a work published on Nature Immunology

role of histone demethylase PHF8 in one of the cancer cell

(Impact Factor 19.4). The financial support from

migration mechanisms: epithelial to mesenchymal transition.

Carver Trust was acknowledged in this publication.

This is a process by which epithelial cells lose their cell polarity and cell to cell adhesion properties and gain the

2016 was a fruitful year for Dr. Qi: Dr. Qi published two

ability to invade, thus initiating the spread of cancer cells

papers as senior author and one as a co-author. These

to areas outside of the original tumor. Specifically, Dr. Qi’s

publications quickly exposed Dr. Qi’s research expertise

lab used genome-wide gene expression profiling (RNA-seq)

in cancer epigenetics, leading to service as an ad hoc

to understand which genes have been turned on at different

reviewer of scientific manuscripts submitted to the journal

stages of breast cancer. This analysis revealed how PHF8

Oncotarget. Dr. Qi was also invited to contribute a book

regulates gene expression in the context of breast cancer

chapter on cancer epigenetics to be published in 2017. Dr.

progression. This work was published in a prestigious journal

Qi submitted an application for federal funding (R01) in

Nucleic Acids Research (Impact Factor 9.2). The financial support

2016 and generated preliminary data for the second R01

from Carver Trust was acknowledged in this publication.

application to be submitted in June, 2017. Moreover, Dr. Qi’s mentorship is getting rewarded as his PhD trainee

Second, during the course of the project described above, Dr.

(Peterson Kariuki Maina) will graduate and go to medical

Qi and a new postdoctoral fellow Dr. Qi Liu discovered an

school in 2017. Furthermore, Dr. Qi and his lab staff are

essential functional link between PHF8 and ERBB2/HER2,

preparing three manuscripts for publication in 2017.

which is a well-known oncogene amplified or over-expressed in 30 percent of breast cancers. This project led to a new internal

In 2016, Qi’s lab published three papers: two papers as

grant to Dr. Qi’s lab from the Holden Comprehensive Cancer

correspondence author in Nucleic Acids Research (Impact Factor:

Center, Breast Cancer Research Group, University of Iowa.

9.2) and Oncotarget (Impact Factor: 5.0), respectively; one

The preliminary data also built a base for a federal funding

co-author paper in Nature Immunology (Impact Factor: 19.4).

(R01) application in October 2016. Currently, this project is ongoing and Dr. Qi aims to publish this work in 2017. Third, in the prostate cancer studies, the Qi lab has discovered a unique regulation pattern of a cluster of epigenetic factors during neuroendocrine differentiation (NED) toward the development of castration resistant prostate cancer (CRPC), a lethal form of prostate cancer. The histone demethylases PHF8 and KDM3A together with well-known epigenetic factors such as EZH2, TOP2A, and HMGB2 go through down-regulation during NED and upregulation in CRPC cells. Dr. Qi and his PhD student Peterson Kariuki Maina further discovered that PHF8 plays a critical role in the development of CRPC cells. This work was published on Oncotarget (Impact Factor 5.0), a well-known journal in cancer research. The financial support from Carver Trust was acknowledged in this publication. Moreover, Dr. Qi and Mr. Maina are preparing a manuscript for publication in 2017. Dr. Qi will also use the preliminary data generated from this

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PUBLICATIONS

1. Shao P, Liu Q , Maina PK, Cui J, Bair TB, Li T, Umesalma S, Zhang W and Qi HH. Histone demethylase PHF8 promotes epithelial to mesenchymal transition and breast tumorigenesis. Nucleic Acids Research. 2016, Nov 29. pii: gkw1093, Epub ahead of print.

2. Maina PK, Shao P, Liu Q , Fazli L, Tyler S, Nasir M, Dong X and Qi HH. c-MYC drives histone demethylase PHF8 during neuroendocrine differentiation and in castration-resistant prostate cancer. Oncotarget. 2016, Sep 28. doi:10.18632/oncotarget.12310.

3. Xing S, Li F, Zeng Z, Zhao Y, Yu S, Shan Q , Li Y, Phillips FC, Maina PK, Qi HH, Liu C, Zhu J, Pope RM, Musselman CA, Zeng C, Peng W, Xue HH. Tcf1 and Lef1 transcription factors establish CD8(+) T cell identity through intrinsic HDAC activity. Nat Immunol. 2016 Jun;17(6):695-703. doi:10.1038/ni.3456. Epub 2016 Apr 25. PMCID: PMC4064003

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

HANK QI, MD, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$62,731

$0

Other (1.7%) Lab Supplies/ Animals (9.8%)

Personnel (88.5%)

Expenses Personnel $55,541 Lab Supplies/Animals $6,152 Other $1,038 Total Expenses $62,732 Ending Balance

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$0

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JANICE ROBERTSON, PHD Dr. Janice Robertson’s research focuses ASSISTANT PROFESSOR, DEPARTMENT OF

MOLECULAR PHYSIOLOGY AND BIOPHYSICS

on the following question—why do greasy membrane proteins choose to interact with their greasy protein partners over the similarly greasy lipid membrane? This reaction is central to understanding how membrane proteins fold in cell membranes, which remains a fundamental yet poorly understood phenomenon of self-assembly in living systems. Driving forces of greasy protein folding in greasy lipid membranes Funds from the Carver Trust were used to support key personnel on the project including an assistant research scientist, a technician/research assistant, a postdoctoral fellow and partial salary support for Dr. Robertson. In addition, funds were used to purchase experimental supplies including lipids, detergents, fluorophores, chemicals, and DNA oligonucleotides. This past year, the Robertson laboratory completed its project on measuring the equilibrium dimerization free energy of the large ClC-ec1 chloride/proton antiporter in lipid bilayers, the results of which are published in the highly respected journal eLife. This is the first study of its kind that shows that large membrane protein complexes in membranes follow the same thermodynamic principles as proteins in water. To study this, the Robertson lab used the innovative and cutting edge approach of single-molecule fluorescence microscopy, to

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inspect the protein at densities far lower than that which exists in the cell. At these low levels, they could directly observe the complexes falling apart into isolated monomers, as was predicted if equilibrium dimerization was occurring. With this, they could measure the full monomer to dimer reaction as a function of the density in the membrane, and extract the free energy, i.e. the intrinsic stability, of the dimer complex. In addition, they were able to show that insertion of bulky tryptophan residues at the dimerization interface result in a quantifiable destabilization in the free energy, laying down the first steps in dissection the protein vs. lipid driving forces that influence membrane protein stability in lipid bilayers. The lab is currently preparing a Research Advance article to be submitted to eLife to follow up on this work. This will report on recent results showing that dimerization can be measured without knowledge of the liposome size distributions using a cross-linked dimer control. In addition, it will verify the equilibrium nature of the measurements by demonstrating subunit exchange by single-molecule FRET. Also, a manuscript is in preparation that describes the theoretical modeling of the Poisson-distribution describing the probabilities of subunit capture into liposomes, and this will be submitted to the Journal of General Physiology later this year. Significant progress has been made on Aim I, which asks the question—are the protein side chains interacting with each other by van der Waals interactions (i.e. molecular Velcro) to stabilize the dimer state? The Robertson lab has constructed a large library of dimerization mutants where the native amino acid has been replaced with a smaller alanine residue, creating empty pockets that remove interactions between the protein surfaces. All constructs have been verified to be folded by functional studies, and now their dimerization free energy is being measured by the established photobleaching approach. These studies have led to the discovery that a single subtractive mutant of a leucine to alanine, leads to the destabilization of the dimer complex in a similar amount to the addition of a single bulky tryptophan. In other words, removing a tiny amount of the Velcro has the same impact as inserting a steric wedge. This was a surprising discovery, and it either means that protein interactions play a significant role, or that they have identified a molecular hot-spot in dimer stabilization. Experiments are ongoing to identify which of these opposing hypotheses are supported. Finally, in order to quantify the exact contribution of the van der Waals interactions, a measurement of the size of the pockets created must be

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made. To do this, the Robertson Lab is working toward obtaining high-resolution crystal structures of the mutant constructs, as well as modeling the mutations using computer simulations. These studies are nearly completed and the manuscript is expected to be published in the upcoming year. Significant progress has also been made on Aim II— investigating the role of hydrophobic mismatch on ClCec1 dimer stability. The Robertson lab made constructs with elongated helices at the dimerization interface, however, the final construct did not express. With that, efforts have focused on studying the change in the lipid structure near the dimerization interface by computer simulations of the monomer and dimer using the coarsegrained MARTINI force-field, which demonstrates a 10 Å thinning of the membrane. To experimentally validate these models, the Robertson Lab has also carried out neutron scattering and x-ray scattering measurements that report on the bilayer thickness changes directly. A manuscript is currently in preparation that reports on these results and will be submitted later this year. In 2016 Dr. Robertson received a R01 grant from the National Institute of General Medical Sciences that will continue the work that has been established with Carver Trust Young Investigator Award. This research was well received by her peers, scoring in the top fourth percentile of all R01 proposals. In addition, Dr. Robertson was awarded beamline time at the NIST Neutron beam source, which will allow for further investigation of the changes in membrane structure related to ClC-ec1 dimerization. Dr. Robertson was nominated to join the Editorial Advisory Board at the Journal of General Physiology, and at the University of Iowa, she was selected to be the Director of the

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Seminar Series of the Department of Molecular Physiology and Biophysics, and elected as the Assistant Professor level member of the Research Advisory Faculty Committee to the Office of the Dean of the Carver College of Medicine. She was a co-organizer of the 4th Midwestern Single-Molecule Workshop held at the University of Iowa, and was selected to co-organize the 72nd Annual Meeting of the Society of General Physiologists, on the topic of “Molecular Physiology of the Cell Membrane: an Integrative Perspective from Experiment and Computation” in 2018. In 2017 she will participate as an ad hoc reviewer of the Biochemistry and Biophysics of Membranes (BBM) peer review committee at NIH. Finally, Dr. Robertson has been invited to speak at numerous seminar series and conferences, including the University of Delaware Membrane Protein Symposium, NINDS/ NIH Membrane Protein Interest Group, Department of Biochemistry at the University of Missouri, Mechanisms of Membrane Transport Gordon Conference, Cold Spring Harbor Asia Conference on Membrane Proteins and the Single Molecule Membrane Protein Dynamics Symposium at the 61st Annual Meeting of the Biophysical Society.

PUBLICATIONS

1. Chadda R, Krishnamani V, Mersch K, Wong J, Brimberry M, Chadda A, Kolmakova-Partensky L, Friedman LJ, Gelles J & Robertson JL (2016). The dimerization equilibrium of a ClC Cl-/H+ antiporter in lipid bilayers. eLife 2016; 10.7554/ eLife.174382016.

2. Chadda R, Robertson JL (2016). Measuring Membrane Protein Dimerization Equilibrium in Lipid Bilayers by Single-Molecule Fluorescence Microscopy. Methods Enzymol. 2016;581:53-82

3. Leisle L, Chadda R, Lueck JD, Infield DT, Galpin JD, Krishnamani V, Robertson JL & Ahern CA (2016). Cellular encoding of Cy dyes for single-molecule imaging. eLife 2016; 10.7554/eLife.19088.

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

JANICE ROBERTSON, PHD Balance Forward

$54,347

Revenue Disbursements to UI Total Revenue

$144,124

Expenses Personnel $126,244 Lab Supplies $23,809 Travel $3,485 Other $6,603 Total Expenses $160,142 Ending Balance

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Travel (2.2%)

Other (4.1%)

Personnel (78.8%)

Lab Supplies (14.9%)

$38,329

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JULIEN A. SEBAG, PHD Dr. J u l i e n Sebag’s research focuses on understanding the role of the Melanocortin ASSISTANT PROFESSOR, DEPARTMENT OF

MOLECULAR PHYSIOLOGY AND BIOPHYSICS

Receptor Accessory Protein 2 (MRAP2) in the control of food intake and energy expenditure through the regulation of hormones and neuropeptide receptors.

Role and Mechanism of MRAP2 Mediated Regulation of Prokineticin Receptors Funds from the Roy J. Carver Charitable Trust were used in 2016 to develop new animal models to demonstrate the role of MRAP2 in the regulation of the prokineticin receptor 1 (PKR1) in-vivo and establish the importance of those proteins in the regulation of food intake. Specifically, funds from the Carver Trust were used to follow up on the identification of MRAP2 as an inhibitor of PKR1 trafficking and signaling in-vivo, the Sebag Lab developed several new mouse models using the state of the art CRISPR/Cas9 gene editing technology. The first model is a mouse that allows the detection of PKR1 in mice using antibodies. This was achieved by inserting an HA tag at the start of the endogenous PKR1 gene. Using this mouse the Sebag lab is now able to map and quantify PKR1 expression in the brain and study PKR1 localization and trafficking in-vivo. The second model allows the localization and detection of MRAP2 in-vivo and was generated by inserting the V5 tag at the end of the endogenous MRAP2 coding sequence. This model allowed the Sebag lab to map MRAP2 in the brain and determine that MRAP2 expression is changed by feeding and fasting. In addition, breeding those two types of mice allowed the production of animals in which both PKR1 and MRAP2 can be identified and permitted to convincingly demonstrate that MRAP2 and

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

JULIEN A. SEBAG, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$40,053

$129,812

Other (3%) Lab Supplies (25.6%)

Personnel (71.4%)

Expenses Personnel $75,012 Lab Supplies $26,895 Other $3,142 Total Expenses $105,049 Ending Balance

$64,816

PKR1 are expressed in a same population of neurons.

Funds from the Carver Trust allowed the Sebag laboratory

The Sebag lab also developed mouse models that allow

to generate a large amount of high quality results that

the targeted deletion or overexpression of MRAP2

have been published in Elife. The funds also permitted

to test the effect of modulating the expression of this

the acquisition of significant preliminary results, thus

protein on PKR1 signaling in-vivo and on feeding behavior.

allowing the submission of an R01 grant (results in March 2017). The findings associated with this project were

Behavioral studies were conducted to determine if the

presented at several international conferences and were

interaction of MRAP2 with PKR1 in-vivo blocks the

received with excitement by the scientific community. Dr.

decrease in food intake caused by the activation of PKR1 by

Sebag was was selected to submit an application for

its ligand PK2. For this, WT and mice lacking MRAP2

the Pew Biomedical Scholars fellowship. In addition, he

were cannulated in the brain and injected with vehicle

was invited to contribute a review on MRAP proteins

or PK2. Results demonstrated that the decrease in food

for BBA—Molecular Basis of Disease. In preparation.

intake caused by PK2 injection was significantly enhanced in mice lacking MRAP2, thus proving the inhibitory action of MRAP2 on PKR1 in-vivo. The Sebag lab also

PUBLICATIONS

showed that, unlike previously suggested by other groups, the role of PKR1 on the regulation of food intake does

1. Chaly AL, Srisai D, Gardner EE, Sebag JA. The Melanocortin

not depend on the activation of the central melanocortin

Receptor Accessory Protein 2 promotes food intake

system since PK2 injection in mice with a deletion of

through inhibition of the Prokineticin Receptor-1. Elife.

MC4R, the main melanocortin receptor for the regulation

2016 Feb 1;5. pii: e12397. doi: 10.7554/eLife.12397.

of food intake, caused a significant decrease in feeding.

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RAJAN SAH, MD, PHD Dr. Rajan Sah’s research focuses on the ASSISTANT PROFESSOR, DEPARTMENT OF INTERNAL

MEDICINE—CARDIOVASCULAR MEDICINE

question of whether cells can sense their size and regulate cellular growth and

ASSISTANT PROFESSOR, DEPARTMENT OF

metabolism accordingly. This question has

broad fundamental implications in biology,

MOLECULAR PHYSIOLOGY AND BIOPHYSICS

health, and disease. To this end, the Sah laboratory is studying the role of a volumesensing molecule (SWELL1) in fat cells to determine how it may regulate fat cell size in the setting of obesity. Tuning fat cell size and obesity through SWELL1 Funds from the Roy J. Carver Charitable Trust were used to support a postdoctoral fellow (Yanhui Zhang) and a research assistant in addition to supporting mouse costs and supplies (antibodies, enzymes, molecular kits) for the above project. As a result, these funds contributed significant preliminary data to supporting multiple grant applications and awards (see details below); to training of postdoctoral fellows and students (including high school students); to establishing collaborations in novel directions; and to generating a manuscript and a book chapter that are likely to be published in 2017. The Carver Trust funds supported further development of molecular tools and mouse models to study SWELL1 in fat cells, including novel viral methods and the use of the emerging CRISPR/cas9 gene-editing technology to manipulate gene expression of fat cells both in mice and in a dish. These advances and the preliminary data that arose

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from them supported multiple grant applications, including: 1) an American Heart Association Postdoctoral Award (Yanhui Zhang), 2) a Center for Hypertension Pilot Grant; 3) a Diabetes Research Center (DRC) Pilot and Feasibility Grant (beta-cell biology), 4) a Carver Trust Collaborative Research Grant, 5) a second R01 application (beta-cell biology), 6) an American Heart Association Postdoctoral Award (Susheel Gunasekar, endothelial SWELL1), and 7) and American Heart Association Grant-in-Aid (beta-cell biology). Yanhui Zhang’s American Heart Association Postdoctoral Award received the top score (0.01 percentile) and was awarded in July 2016 ($100,000 over two years). The Center for Hypertension Grant was also awarded ($60,000), started in August 2016, and represents a successful collaboration between the Sah and Irani laboratories, based on work spawned by Carver Trust funds. The DRC Pilot grant just missed being funded, but we expect a resubmission will do well in 2017. The Carver Trust Collaborative Research Grant was submitted late in 2016 and was awarded in February 2017. This grant explores an off-shoot project from the funded Carver Trust Award related to SWELL1 and pancreatic beta-cell biology. A second R01 application is in the pipeline for Feb. 5, 2017 submission. A second postdoctoral AHA grant on the role of SWELL1 in endothelial biology based on Hypertension grant work is in the pipeline for Feb. 10, 2017 submission. An AHA grant-inaid application is in the pipeline for Feb. 2017 submission. Overall, the first year and a half of Carver Trust funding has contributed to an R01 ($2,061,908 total), GAP Commercialization award ($75,000), Commercial Funding (Kemin, $45,000), AHA Postdoctoral award ($100,000) and HTN pilot grant ($60,000). Accounting for start-up funds, this represents a return on investment of $1,341,928/~$188,500 = ~712 percent. In addition, Carver Trust funding allowed for the pursuit of off-shoot projects and collaborations, including examining the role of SWELL1 in myocardium (Sah lab only), SWELL1 in adipose inflammation (Sah lab only), SWELL1 in endothelial cell function (Sah/Irani, UIowa), SWELL1 in pancreatic beta cell function (Sah/Norris/Imai/Stephens), SWELL1 microglia function (Sah/Wu, Rutgers), SWELL1 leptin regulation (Sah/Friedman, Rockefeller) and SWELL1 structure-function (Sah/Robertson, UIowa). In the latter collaboration, it is notable that both Sah and Robertson

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laboratories hold Carver Young Investigator Awards. Indeed, this example highlights the synergistic use of Carver Tr u s t funding to combine state-of-the art techniques, in this case single-molecule super-resolution fluorescence microscopy (Robertson), with the fundamental understanding of a novel volume-sensing molecule, SWELL1 (Sah). Finally, the manuscript currently in review with Nature Cell Biology, and largely supported by Carver Trust funding, entitled “SWELL1 is a regulator of adipocyte size, insulin signaling, and glucose homeostasis” delineates a novel role for the recently discovered swell-activated channel, SWELL1, in fat cell (adipocyte) biology. The Sah laboratory shows that this molecule is activated in adipocytes of obese mice and humans, and is required for regulating fat content and sugar metabolism through its effects on the insulin signaling pathway. They identify molecular partners and interactions responsible for these biological effects. They also show that eliminating SWELL1 from adipocytes limits adipocyte size and reduces fat mass, but worsens blood sugar levels. In sum, these studies identify SWELL1 as a cell-autonomous sensor of adipocyte size that regulates adipocyte growth, insulin sensitivity, and glucose tolerance in the setting of obesity. As SWELL1 is active in other tissues and its downstream signaling ubiquitous and fundamentally intertwined with numerous signaling pathways, t he Sah laboratory anticipates that SWELL1 signaling will be physiologically and pathophysiologically important in a multitude of different tissues and disease states (thus the extensive collaborative projects initiated and listed above). Accordingly, this work will contribute significantly, not only to the field of adipocyte biology, but to cell biology at large, by inspiring further investigation into “mechano-tuning” of intracellular signaling via SWELL1 in numerous other cell types.

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In 2016 Dr. Sah spoke at the Naomi Berrie Diabetes Center at Columbia University, at the Fraternal Order of the Eagles Diabetes Center (U Iowa), and the Pulmonary Research Conference (U Iowa). Dr. Sah co-directed the Carver College of Medicine Research Day (inviting Drs. Feng Zhang and George Daley) and gave a short talk. Dr. Sah and postdoctoral fellow Yanhui Zhang presented their research at The Society of General Physiology (SGP) meeting in Woodshole, MA, in September 2016 and attended the Keystone Symposium on Obesity and Diabetes in January 2017. Dr. Sah’s first postdoctoral fellow Yanhui Zhang has landed a faculty position in China based on her Carver Trust-funded project, which represents a true success story in mentorship and career development. Finally, Dr. Sah’s Carver Trust-funded project provided preliminary data upon which he built an NIH R01 application that was awarded by the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) in April 2016. Dr. Sah was also awarded a GAP Commercialization Award to further develop a technology that the Sah laboratory has developed and is patenting. Moreover, Dr. Sah established a partnership with industry to explore the biological activity of these industry compounds as well.

PUBLICATIONS

1. Zhang Y, Xie L, Gunasekar S, Tong D, Mishra A, Gibson W, Wang C, Fidler T, Marthaler B, Klingelhutz A, Abel E.D, Smith J, Samuel I, Cao L, and Sah R. SWELL1 is a regulator of adipocyte size, insulin signaling and glucose homeostasis Nature Cell Biology (in review, 2/3 reviewers accepted)

2. Zhang Y, Tong D, Misra A, Xie L, Samuel I, Smith JK, Sah R. “Isolation and Patch-­‐Clamp of Primary Adipocytes.” In “Thermogenic Fat: Methods and Protocols; Methods Molecular Biology (Vol 1566)”, Eds. Jun Wu, Springer-­Nature ISBN: 978-­‐1-­‐4939-­‐6819-­‐0, 2017.

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January 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

RAJAN SAH, MD, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$53,148

$137,218

Other (23%)

Personnel (38.7%)

Expenses Personnel $34,694 Lab Supplies/Animals $34,318 Other $20,586 Total Expenses $89,599 Ending Balance

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$100,768

Lab Supplies/ Animals (38.3%)

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RYAN L. BOUDREAU, PHD Dr. Ryan Boudreau’s research focuses on ASSISTANT PROFESSOR, DEPARTMENT OF INTERNAL

MEDICINE—CARDIOVASCULAR MEDICINE

understanding how cardiac cells regulate their protein-coding RNAs to control when and where specific proteins are expressed under normal conditions and in disease. His work has the potential to uncover new RNA-based medicines for heart disease and improve our understanding of how human genetic variations may contribute to disease outcomes by altering cardiac RNA regulatory mechanisms.

Systematic characterization of post-transcriptional gene regulatory networks in cardiac disease Funds from the Roy J. Carver Charitable Trust were used to: 1) make substantial headway toward identifying the hundreds of cellular proteins that control RNA regulation in heart tissues; 2) discover new RNA elements that control cardiac gene regulation in the setting of ischemic cardiomyopathy; and 3) identify two novel RNA regulators that are suppressed in failing human and rodent hearts. Our genomes encode thousands of unique proteins needed for our cellular makeup and function. Each protein-coding gene is first transcribed into individual messenger RNAs (mRNAs) that are then precisely regulated to control when and where they are translated into proteins. This regulation, in part, is mediated by a complex network of hundreds of

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RNA-binding proteins (RBPs) that recognize specific sequence codes within the mRNAs. In the heart, these processes remain largely understudied, and Dr. Boudreau aims to employ high-throughput and innovative wet-lab and computational approaches to expand our knowledge of how cardiac cells regulate their mRNAs and how these processes are disrupted in heart disease. Specifically, as supported by the Carver Trust Young Investigator Award 2016, research investigators in the Boudreau lab will: 1) globally identify the cardiac proteins that engage mRNAs in human and mouse heart tissues; 2) systematically identify the mRNA sequence codes that associate with gene expression changes in diseased hearts; and 3) determine which RBPs are dysregulated in failing hearts from human patients and rodent models. The resulting data will provide critical information and resources to advance our understanding of the biological- and disease-relevance of

analyses to identify many other RBPs present with in these samples (anticipated data in 2017). Pilot studies were done using mouse hearts; human samples will be tested soon.

cardiac mRNA regulation. In addition, this work will support

Aim 2 progress: identify the mRNA sequence

our future efforts to identify genetic risk factors contributing

codes that confer gene regulatory function in

to heart failure and to assess whether therapeutic targeting of

heart

RNA regulatory pathways can improve disease outcomes. Aim 1 progress: globally identify the cardiac proteins that engage mRNAs in cardiac tissues Funds provided by the Carver Trust supported research staff, as well as the cost of supplies and animals needed for obtaining and processing cardiac tissues samples that were used for adapting and troubleshooting established protocols for fishing out the mRNA-bound proteome (i.e. RBPs). Notably, these protocols have predominantly been developed for applications in cell culture systems, and performing these procedures on solid tissue samples poses several hurdles that require optimization. Through a series of experiments, Dr. Boudreau’s group efficiently identified the

Previously, Dr. Boudreau’s lab used bioinformatic means to analyze gene expression changes (at the RNA level) that occur in failing hearts, both humans and rodents, with the goal of identifying RNA regulatory elements (RREs) that associate with observed changes. In 2016, these methods have been applied to several other comparable datasets to identify additional candidate RREs for further testing. Through follow-up wet-lab experiments, lab investigators have identified at least a handful of RREs that impart gene regulatory functions in cell-based reporter assays. Continued efforts are ongoing to test additional candidate RREs and identify their RBP binding partners using the proposed biochemical-based “pull-down” approaches.

pitfalls of applying this method to cardiac tissues and devised

Aim 3 progress: determine which RBPs are

strategies to overcome each of them. One notable obstacle

dysregulated in failing hearts

was the fact that heart tissues are loaded with mitochondria (tiny organelles that supply energy to our cells; the heart requires a lot of power to keep beating). Unfortunately, these mitochondria were “gobbing up” our experimental system and blocking our ability to pull-down cardiac mRNAs and their associating proteins. Lab investigators employed standard means to remove the mitochondria from the samples, and this was critical for the protocol to work. The lab has recently generated “pull-down” samples, in which they have confirmed the presence of known RBPs, and they are gearing up to perform a preliminary round of proteomic

YO U N G I N V E S T I G AT O R AWA R D S

In 2016, the Boudreau lab used funds provided by the Carver Trust to determine the abundance of candidate RBPs in a panel of cardiac tissues, including normal control samples and failing hearts derived from human subjects and rodent models. Technicians in the Boudreau lab employed surgical techniques to induce heart failure in a cohort of mice, in order to generate the needed samples for these studies. Human heart failure tissue samples were obtained from our collaborators at University of Pennsylvania. Funds were also used to purchase supplies and molecular reagents needed to detect and measure the amount of specific RBPs within

149


the biological samples. Using standard approaches for protein quantitation, the Boudreau lab has discovered two novel RBPs that are significantly down-regulated in both human and rodent failing hearts. These data are exciting and suggest the potential for translational studies (mouseto-man). Future research in mice will need to determine the potential disease-relevance of these RBPs in heart. Dr. Boudreau was awarded a pilot grant from the University of Iowa Fraternal Order of the Eagles Diabetes Research Center and received an Inventor Award from the University of Iowa Research Foundation, in recognition of innovation leading to patents and intellectual property licensing. In addition, he published his first manuscript as a corresponding author (i.e. Principle Investigator).

PUBLICATIONS

1. R.M. Spengler, X. Zhang, C. Cheng, J.M. McLendon, J.M. Skeie, C. Cheng, F.L. Johnson, B.L. Davidson, and R.L. Boudreau. Elucidation of transcriptome-wide microRNA binding sites in human cardiac tissues by Ago2 HITS-CLIP. (2016) Nucleic Acids Research 44(15):7120-7131.

150

2 0 1 7 A N N U A L R E P O R T TO T H E C A R V E R T R U S T


May 1, 2016 — December 31, 2016

FINANCIAL SUMMARY

RYAN BOUDREAU, PHD Balance Forward Revenue Disbursements to UI Total Revenue

$0

$148,421

Other (7.4%) Lab Supplies/ Animals (35.9%)

Personnel (56.7%)

Expenses Personnel $53,975 Lab Supplies/Animals $34,205 Other $7,085 Total Expenses $95,265 Ending Balance

YO U N G I N V E S T I G AT O R AWA R D S

$53,156

151



FINANCIAL SUMMARY

153


$63 MILLION GIFT CARVER COLLEGE OF MEDICINE CAMPUS IMPROVEMENTS & CARVER BIOMEDICAL RESEARCH BUILDING To support improvements and the Carver Biomedical Research Building. Original Pledge: $10,000,000 2005 2003

2002

2006

2004

2007 Pledge Remaining:

$0 pledge used

ROY J. CARVER RESEARCH PROGRAMS OF EXCELLENCE To support five research programs. Original Pledge: $15,000,000 2016 2002

2003

2004

2005

2006 2007

2008

2009 2010

2011

2012

2013 2014 2015 Pledge Remaining:

$850,000 pledge used

pledge used FY2016

CARVER TRUST RESEARCH FACILITIES SUPPORT To support core research facilities construction, renovation and operations. 60% to support long-term operational needs of existing and future core facilities. 40% to support capital project grants. Original Pledge: $10,000,000 2007 2006 2002

2005

2009

2010

2011

2013 Pledge Remaining:

$0

pledge used

Original Pledge

FY2002

FY2003

FY2004

FY2005

FY2006

FY2007

CCOM Campus Improvements & Carver Biomedical Research Building

10,000,000

2,500,000

2,300,000

1,250,000

13,280

2,681,402

1,255,318

Roy J. Carver Research Programs of Excellence

15,000,000

1,000,000

1,000,000

1,000,000

1,000,000

1,000,000

1,000,000

Carver Trust Research Facilities Support

10,000,000

300,000

0

0

3,436,720

130,522

134,437

Carver Trust Individual Research Grants

8,000,000

500,000

300,000

403,268

392,785

292,441

200,000

Carver Trust ResearchRelated Endowments

20,000,000

500,000

0

2,000,000

0

2,000,000

2,000,000

$63,000,000

$4,800,000

$3,600,000

$4,653,268

$4,842,785

$6,104,365

$4,589,755

Total

154

2 0 1 7 A N N U A L R E P O R T TO T H E C A R V E R T R U S T


CARVER TRUST INDIVIDUAL RESEARCH GRANTS* $200k per year to support Medical Research Initiative Awards Program. Remaining will be awarded as 1-3 year grants to individual research programs on a per-invitation basis. Original Pledge: $8,000,000 2005

2003 2002

2007 2009

2006 2008

2004

2010

2011* 2012

2015

2014

2013

Pledge Remaining:

$906,774 pledge used

CARVER TRUST RESEARCH-RELATED ENDOWMENTS To establish research-related endowments to be associated with specific departmental or division headships and professorships. Original Pledge: $20,000,000

2002

2004

2006

2007

2008 2010

2014

2012

2011

Pledge Remaining:

$0 pledge used

TOTAL Pledge

FY2002 - FY2015 Payments

FY2016 Payments

Remaining:

$1,756,774

* Per Carver Trust, pledge reduced by $1,679 due to overpayment on Research Facilities pledge

FY2008

FY2009

FY2010

FY2011

FY2012

FY2013

FY2014

FY2015

0

0

0

0

0

0

0

0

1,000,000

1,000,000

1,000,000

1,000,000

1,000,000

1,000,000

750,000

700,000

0

2,000,000

1,000,000

1,000,000

0

2,000,000

0

0

200,000

200,000

504,599

781,490

376,900

591,692

578,449

1,026,931

2,000,000

0

1,000,000

3,000,000

2,000,000

0

2,000,000

0

3,500,000

20,000,000

$3,200,000

$3,200,000

$3,504,599

$5,781,490

$3,376,900

$3,591,692

$3,328,449

$1,726,931

4,942,992

61,243,226

FINANCIAL SUMMARY

FY2016

0

700,000

Total Payments

10,000,000

14,150,000

10,001,679

$742,992

7,091,547

155


EN D OW MEN TS JULY 1, 2015 THROUGH JUNE 30, 2016

Grant #

01-224

01-224

Account Name Roy J. Carver Chair in Psychiatry and Neuroscience Roy J. Carver Professorship in Molecular Physiology and Biophysics

Gift/pledge/other

Market Value

Balance

income

Appr / (Depr)

0

2,000,000

0

0

1,500,000

0

01-224

Roy J. Carver Chair in Molecular Genetics

3,425,329

0

(42,588)

01-224

Roy J. Carver Chair in Neuroscience — III

2,054,272

0

(24,702)

01-224

Tissue Procurement Facility

2,154,376

0

(26,668)

2,309,173

0

(28,490)

01-224

Roy J. Carver Chair in Neuroscience — Deparment of Psychiatry

01-224

Roy J. Carver Chair in Neuroscience

2,057,067

0

(25,623)

01-224

Roy J. Carver Chair in Biochemistry

2,327,568

0

(28,769)

01-224

Proteomics Facility

2,327,568

0

(28,769)

01-224

Genomics Facility

2,781,732

0

(34,564)

01-224

Roy J. Carver Chair in Hypertension Research

2,109,874

0

(26,216)

01-224

Roy J. Carver Chair in Pulmonary Research

1,897,145

0

(23,572)

01-224

Roy J. Carver Chair in Molecular Medicine

2,065,337

0

(25,663)

747,413

200*

(6,994)

1,790,912

2,000*

(20,248)

1,529,256

0

(19,001)

1,529,256

0

(19,001)

3,351,086

0

(41,176)

01-224

01-224

99-165

99-165

97-9

Nicholas P. Rossi Professorship — Cardiothoracic Surgery Francois Abboud Chair in Internal Medicine Roy J. Carver Biomedical Research Chair in Physiology & Biophysics Roy J. Carver Biomedical Research Chair in Internal Medicine/Welsh Carver Molecular Ophthalmology Laboratory Fund

87-5

Roy J. Carver Chair in Internal Medicine

2,935,979

0

(36,481)

01-241

James A Clifton Chair in Gastroenterology

1,428,708

1,500*

(1,380)

$38,822,051

3,503,700

(459,905)

Total

156

Beginning

2 0 1 7 A N N U A L R E P O R T TO T H E C A R V E R T R U S T


Interest

Market Value

Management

Disbursements

Ending

Income / Payout

Allocated to Payout

Fees

(To UI)

Balance

0

0

0

0

2,000,000

0

0

0

0

1,500,000

156,407

(156,407)

(32,065)

(156,097)

3,194,579

90,720

(90,720)

(18,599)

0

2,010,971

92,081

(92,081)

(20,207)

(91,898)

2,015,603

93,646

(93,646)

(21,690)

(93,460)

2,165,533

96,456

(96,456)

(19,241)

(96,264)

1,915,939

97,179

(97,179)

(21,845)

(96,986)

2,179,968

97,179

(97,179)

(21,845)

(96,986)

2,179,968

125,840

(125,840)

(26,048)

(125,590)

2,595,530

95,446

(95,446)

(19,756)

(95,257)

1,968,645

85,823

(85,823)

(17,765)

(85,653)

1,770,155

93,432

(93,432)

(19,339)

(93,246)

1,927,089

25,432

(25,432)

(5,278)

(32,000)

703,341

76,395

(75,797)

(17,130)

(130,980)

1,625,152

69,180

(69,180)

(14,320)

(69,043)

1,426,892

69,180

(69,180)

(14,320)

(69,043)

1,426,892

149,914

(149,914)

(31,031)

(186,719)

3,092,160

132,818

(132,818)

(27,492)

(132,554)

2,739,452

58,611

(48,252)

(22,916)

(39,017)

1,377,254

1,705,739

(1,694,782)

(370,887)

(1,690,793)

39,815,123

*Includes contributions/adjustments from other donors

FINANCIAL SUMMARY

157


EN D OW MEN TS JULY 1, 2016 THROUGH DECEMBER 31, 2016

Grant #

01-224

01-224

Account Name Roy J. Carver Chair in Psychiatry and Neuroscience Roy J. Carver Professorship in Molecular Physiology and Biophysics

Gift/pledge/other

Market Value

Balance

income

Appr / (Depr)

2,000,000

0

107,610

1,500,000

0

80,707

01-224

Roy J. Carver Chair in Molecular Genetics

3,194,579

0

169,749

01-224

Roy J. Carver Chair in Neuroscience — III

2,010,971

1,000,000

98,459

01-224

Tissue Procurement Facility

2,015,603

0

107,205

2,165,533

0

115,261

01-224

Roy J. Carver Chair in Neuroscience — Deparment of Psychiatry

01-224

Roy J. Carver Chair in Neuroscience

1,915,939

0

101,766

01-224

Roy J. Carver Chair in Biochemistry

2,179,968

0

115,984

01-224

Proteomics Facility

2,179,968

0

115,984

01-224

Genomics Facility

2,595,530

0

137,937

01-224

Roy J. Carver Chair in Hypertension Research

1,968,645

0

104,622

01-224

Roy J. Carver Chair in Pulmonary Research

1,770,155

0

94,073

01-224

Roy J. Carver Chair in Molecular Medicine

1,927,089

0

102,413

703,341

0

27,956

1,625,152

0

82,344

1,426,892

0

75,831

1,426,892

0

75,831

3,092,160

0

164,325

2,739,452

42,241*

145,586

1,377,254

1,000*

37,469

39,815,123

1,043,241

2,061,112

01-224

01-224

99-165

99-165

97-9

Nicholas P. Rossi Professorship — Cardiothoracic Surgery Francois Abboud Chair in Internal Medicine Roy J. Carver Biomedical Research Chair in Physiology & Biophysics Roy J. Carver Biomedical Research Chair in Internal Medicine/Welsh Carver Molecular Ophthalmology Laboratory Fund

87-5

Roy J. Carver Chair in Internal Medicine

01-241

James A Clifton Chair in Gastroenterology

Total

158

Beginning

2 0 1 7 A N N U A L R E P O R T TO T H E C A R V E R T R U S T


Interest

Market Value

Management

Disbursements

Ending

Income / Payout

Allocated to Payout

Fees

(To UI)

Balance

45,000

(45,000)

(10,430)

(22,500)

2,074,680

33,750

(33,750)

(7,823)

(16,875)

1,556,009

78,751

(78,751)

(16,446)

(78,477)

3,269,405

45,678

(45,678)

(9,539)

0

3,099,891

46,363

(46,363)

(10,390)

(46,202)

2,066,216

47,151

(47,151)

(11,173)

(46,987)

2,222,634

48,565

(48,565)

(9,858)

(48,397)

1,959,450

48,930

(48,930)

(11,241)

(48,760)

2,235,951

48,930

(48,930)

(11,241)

(48,760)

2,235,951

63,360

(63,360)

(13,365)

(63,140)

2,656,962

48,057

(48,057)

(10,137)

(47,890)

2,015,240

43,212

(43,212)

(9,115)

(43,062)

1,812,051

47,043

(47,043)

(9,923)

(46,879)

1,972,700

12,806

(12,806)

(2,709)

0

728,588

38,536

(38,187)

(8,758)

0

1,699,087

34,832

(34,832)

(7,347)

(34,711)

1,460,665

34,832

(34,832)

(7,347)

(34,711)

1,460,665

75,482

(75,482)

(15,921)

0

3,240,564

66,874

(66,874)

(14,106)

0

2,913,173

30,585

(24,318)

(11,451)

(39,191)

1,371,248

938,737

(932,121)

(208,320)

(666,542)

42,051,230

*Includes contributions/adjustments from other donors

FINANCIAL SUMMARY

159



COLLEGE LEADERSHIP

161


CARVER COLLEGE OF MEDICINE LEADERSHIP

162

Jean E. Robillard, MD Vice President for Medical Affairs, University of Iowa, Dean

Patricia L. Winokur, MD Executive Dean, Interim Vice Dean for Research, Associate Dean for Clinical and Translational Science

Christopher S. Cooper, MD Senior Associate Dean for Medical Education

Lois J. Geist, MD Associate Dean for Faculty Affairs and Development

Mark J. Hingtgen, MPA Assistant Vice President for Finance

Boyd M. Knosp, MS Associate Dean for Information Technology

Daniel T. Tranel, PhD Associate Dean for Graduate and Postdoctoral Studies

Douglas J. Van Daele, MD Vice Dean for Clinical Affairs, Executive Director, UI Physicians

2 0 1 7 A N N U A L R E P O R T TO T H E C A R V E R T R U S T


Mark C. Wilson, MD, MPH Associate Dean for Graduate Medical Education

David P. Asprey, PhD, PA-C Assistant Dean for Student Affairs and Curriculum

Amy Lee, PhD Assistant Dean for Research

David J. Moser, PhD Assistant Dean for Faculty Affairs and Development

Robert Piper, PhD Director, Core Research Facilities

Rebecca J. Waltman, MBA Director of Facilities Planning and Management

COLLEGE LEADERSHIP

Sherree A. Wilson, PhD Associate Dean for Cultural Affairs and Diversity Initiatives

Steven R. Craig, MD Assistant Dean for Student Affairs and Curriculum

Denise A. Martinez, MD Assistant Dean for Cultural Affairs and Diversity Initiatives

Greg C. Nelson, MA Assistant Dean and Director, Office of Statewide and Clinical Education Programs

Kristi J. Ferguson, MSW, PhD Director, Consultation and Research in Medical Education

Kelley Ashby Director, Continuing Medical Education

163


Office of Research 100 Medicine Administration Building Iowa City, IA 52242-1101 www.medicine.uiowa.edu/research

Levitt Center for University Advancement 1 West Park Road P.O. Box 4550 Iowa City, IA 52242-4550 uiowa-foundation@uiowa.edu www.uifoundation.org