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ALS Research Workshops 2-3 ALS Genetics 4-5 Research Abstracts 5 Safenowitz Fellowships 6-7 Journal News 8

FALL 2013

The TREAT ALS Research Portfolio ™

TREAT ALS™ Research Projects In 2013, The ALS Association awarded grants to 37 researchers in seven countries throughout the world to further the fight against amyotrophic lateral sclerosis (ALS). Researchers funded by these grants will seek to:

flies. In ALS patients, researchers will examine non-mutant variations in known ALS genes for clues to sporadic ALS. Also, a publicly available database will be developed for genomic data of ALS patients, promoting new gene discovery.

Better understand disease mechanisms, including the effects of newly discovered ALS genes and the actions of mutant proteins. New ALS genes have provided new ideas of disease pathogenesis, which researchers will seek to clarify. The mechanism of the FUS gene will be explored in a new mouse model in which the introduced gene is controlled by a conditional promoter, allowing it to be turned off and on. Newly discovered non-standard translation products from the C9orf72 gene will be studied for their potential role in pathogenesis. A histone modifying enzyme will be studied to determine what role it plays in accelerating disease progression. In addition, the mechanisms behind the selective vulnerability of cortical motor neurons will be studied.

Develop biomarkers to track disease progression and explore disease mechanisms. Imaging and biofluids offer the possibility of gaining insight into the progression of disease and revealing new clues to disease mechanisms. Researchers will develop potential ALS biomarkers based on immunoglobulins, PET imaging, MRI and non-standard peptides in C9orf72 patient cells.

Identify new genes that influence ALS risk and affect motor neuron vulnerability. Major disease-causing genes have been identified for ALS, but it is clear the genome contains many other genes that modify risk. Researchers will seek to discover and better understand these genes through genetic screens in fruit

Discover and test potential treatments in cell and animal models. Researchers will develop a drug screen for compounds that stabilize TDP-43 to prevent misfolding; develop potential therapies that reduce toxic aldehydes; develop and test a gene therapy vector for delivery of interfering RNA; and test the therapeutic potential of a compound from the herbal medicine Ashwagandha in mouse models. Use stem cells to model ALS, understand mechanisms and test new therapeutic approaches. Induced pluripotent stem cells (iPS cells) provide the ability to generate patient-specific stem cells for testing disease mechanisms and screening potential therapies. Researchers will

exploit iPS cells to examine differentiation and selective vulnerability of motor neurons in ALS, to develop gene therapies, and to explore the disease mechanisms of mutations in C9orf72 and TDP-43. Accelerate therapy development through testing new agents in ALS patients. Funding will go to support a Phase II study of rasagiline as a potential neuroprotective agent in ALS. Funds will also support continuing development of antisense oligonucleotide (ASO) therapy against mutant SOD1. Previously funded work has shown that administration of ASOs is safe in ALS patients. Now researchers will determine the normal turnover time of SOD1 protein in the cerebrospinal fluid, to develop metrics for efficacy as this therapy moves forward.

Call for Research Abstracts - 5




Research Advances Indicate Exciting Progress The Association’s Global Research Program TREAT ALS™ continues to support a diverse but strategic portfolio of research grants focused on a better understanding of the disease to enable the development of effective therapies. With the significant advances in the last few years in identifying new genes linked to the disease, the investigator community has expanded significantly, exploring the many new avenues that these discoveries have created. This is evident from the increasing number of applicants to our grant program, the increased investment by the pharmaceutical industry in ALS research and the increased number of publications each month relating to ALS. This is extremely encouraging for the field and for a disease in desperate need of an effective treatment that significantly extends and improves the quality of life.

Lucie Bruijn, Ph.D., M.B.A. Chief Scientist The ALS Association

Through the efforts of the investigator community, continued collaboration and brainstorming at meetings and workshops, the field has made significant research advances, which they are beginning to translate in the clinic. The basic scientists are working more closely with clinicians and recognizing the need to validate findings in model systems in the laboratory with human studies. The opportunities created with technologies such as induced pluripotent stem cells, enabling the modeling of disease in cells directly relevant to individual people with the disease, has made this even more feasible.

It is encouraging to see the increasing number of young researchers applying to the Milton Safenowitz Post-Doctoral Fellowship program, and we are pleased to profile the work and research plans of this year’s recipients. Investing in and mentoring the young leaders in ALS research today will ensure growth and advancement in the field in the future. The creativity and enthusiasm of these young individuals fuel ALS research, and we look forward to learning of their progress and accomplishments. -Lucie Bruijn, Ph.D., M.B.A.

RESOURCES ALS mutations database Coriell NINDS DNA repository ALS Epidemiology SOD1 mutant mice, Jackson Laboratory mouse models

SOD1 mutant rats, Taconic

Jackson Laboratories ALS Mouse Repository / Control and SOD1 fibroblasts ALS Untangled ALS Association Research Webinars

The ALS Association Research Workshops Scientific meetings are crucial for bringing together researchers to spur new ideas and new collaborations in their fight against ALS. The Association has sponsored a rich mix of meetings this year, probing aspects of basic biology and pursuing avenues for speeding clinical trials. The Association has also partnered with other groups as part of its strategy to accelerate the search for new treatments and seeks out scientists from other fields to join the effort. Here, we present some highlights from these meetings. Exploring Ideas of Evolution and Development to the Fight Against ALS

In April, The ALS Association brought together more than two dozen scientists from a wide variety of backgrounds to ask whether the patterns of susceptibility seen in motor neurons may be rooted in part in evolution and development. The meeting, funded by The Greater New York Chapter of The ALS Association, took place at the Banbury Center at Cold Spring Harbor Laboratories in Long Island, New York and fostered a wide-ranging discussion among neuroscientists, evolutionary biologists and neurologists. “A key question in ALS is whether the disease takes advantage of differences among motor neurons that have arisen either through evolution of the nervous system or its development during the lifetime,” said ALS Association Chief Scientist Lucie Bruijn, Ph.D., M.B.A., who convened the meeting. “We know these differences exist; what we don’t yet know is what role they may play in susceptibility.” One highlight of the meeting was a discussion of whether the disease process spreads from cell to cell, along networks of neurons that are connected through functional and developmental pathways. William Seeley, M.D., of the University of California at San Francisco, argued that this phenomenon characterizes ALS and most of the other neurodegenerative diseases. “Systems that can be detected in the healthy brain form the basis of degeneration in disease,” he said. John Ravits, M.D., of the University of California at San Diego, proposed instead that spread occurred by physical proximity in the nervous system, with the disease process beginning in a random location, but then moving on to affect neurons adjacent to the site of onset. Continued on page 3


The ALS Association Research Workshops Continued from page 3

An alternative was proposed by Jeffrey Macklis, Ph.D., from Harvard University, who co-chaired the meeting and is an expert on the development of the motor neuron system. “There is not just one type of motor neuron,” he emphasized. In fact, there are hundreds, based on differences in their sizes, shapes, surface markers and developmental history. “This may create selective vulnerability, and the known ALS genes may act on that vulnerability,” he said. There are not yet enough data to know whether one or the other model, or both, are correct. But the implications are significant. “Spread implies that unaffected areas can be protected,” noted Jeremy Shefner, M.D., of SUNY Upstate Medical University in Albany, N.Y. “Selective vulnerability may be harder to protect against.” If the selective vulnerability model is correct, it becomes that much more important to understand the resilience of certain types of motor neurons that remain intact long after other types have degenerated. These include the motor neurons that control eye movements and those that regulate the action of other motor neurons to produce very fine control (so-called gamma motor neurons). Opportunities for Improving Therapy Development in ALS In May 2013, The ALS Association and The Northeast ALS Consortium (NEALS) convened a meeting of clinical researchers, regulatory authorities, industry representatives and patient advocates for a roundtable discussion of ways to improve therapy development in ALS, in light of recent gene discoveries, new disease models and new ideas of pathogenesis. Changes in trial design can play a role in speeding testing of new therapies, including using a centralized Institutional Review Board (IRB) for all clinical trial sites, rather than individual IRBs at each institution. Most important, trials of new drugs should employ pharmacodynamic biomarkers in order to determine if the drug is getting to its target and whether it is altering its target. In that way, even a negative result can be informative for development of other treatments. In addition, clinical biomarkers will be valuable for objectively tracking disease progression and reducing the number of patients needed in a trial.

Workshop participants at Banbury Center at Cold Spring Harbor Laboratories, Long Island, New York.

In August, leading researchers met with people living with ALS and their caregivers at the Team Gleason ALS Summit in New Orleans, sponsored by Team Gleason. The scientific program was organized by ALS Association Chief Scientist Lucie Bruijn, Ph.D., M.B.A. and Jeffrey Rothstein, M.D., Ph.D., Director of the Packard Center for ALS Research. More than 100 attendees brainstormed about new approaches to expedite ALS research and therapy development. Upcoming Meetings

Partnering with Other Organizations

In October, the third annual meeting of the ALS Clinical Research Learning Institute (CRLI), led by Drs. Richard Bedlack and Merit Cudkowicz, will be held in Clearwater Beach, Florida, in conjunction with the annual meeting of the Northeast ALS Consortium (NEALS). The CRLI, sponsored by The ALS Association’s TREAT ALS™ initiative, is an intensive program dedicated to educating patientcaregiver pairs on clinical research and therapy development, while empowering them to be advocates for ALS clinical research, or “Research Ambassadors.”

In June, The ALS Association co-sponsored, with the ALS Society of Canada, the Fourth International Research Workshop on Frontotemporal Dementia in ALS, working to explore the links between ALS and FTD. The meeting organized by Michael Strong, M.D. honored the founder of ALS Society of Canada, Arthur James Hudson, M.D., who sadly passed away in September of this year.

In November, The ALS Association will co-sponsor “RNA Metabolism in Neurological Disease,” a two-day research symposium in conjunction with the annual meeting of the Society for Neuroscience. The symposium will explore the growing understanding of the role of RNA in neurodegenerative diseases, including ALS. For more information, see




ALS Genetics: The View from Everest Camp II

It is also becoming clear that categorization of ALS patients as familial or sporadic based on their family history should not be viewed in absolute terms. The barrier between these two forms of disease is increasing recognized as artificial. This fundamental shift in our understanding of the disease carries significant implications for clinical care of patients and their families, not least of which is genetic counseling.

By Bryan J. Traynor, M.B., M.D., Ph.D., M.M.Sc., M.R.C.P.I., Investigator and Chief of the Neuromuscular Diseases, National Institutes of Health

One does not climb Everest in a single day. Climbers typically spend eight weeks at Everest Base Camp acclimatizing to the altitude before attempting the arduous trek to the top. Even then, the journey is broken by a series of four camps located at progressively higher altitudes, each providing shelter for the night and much needed supplies. The last one before the summit, Camp IV, is so high that climbers can only spend a short time there before making a mad dash for the summit. Unraveling the genetic causes of ALS is a lot like climbing Everest. Each new discovery represents a mere step in the ascent to the top. So where are we right now? The field has made amazing progress and we are sitting pretty in Camp II admiring the amazing views of the Western Cwm and the gently rolling glacial valleys beneath us. We understand the genetic etiology of two thirds of familial ALS and about 10 percent of more common sporadic form of ALS. Driven by technology, genetic discoveries are coming fast and furious with six new genes in the last three years alone.

Everest route map with base camp locations

These discoveries are radically changing how we think about the disease. Clinical, epidemiological and neuropathological data long suggested that ALS and frontotemporal dementia (FTD) overlap. The recent discovery by our group and others of a massive hexanucleotide repeat expansion in C9orf72 demonstrated that a single genetic mutation could underlie both conditions, effectively uniting them into a single disease entity. Other genetic findings have extended the ALS phenotype beyond the central nervous system. Mutations in valosin-containing protein (VCP) are associated with not only ALS and FTD, but also Paget’s disease of the

50s: DNA structure solved 50s: Nerve growth factor (NFG) identified–protective, growth promoting factor for nerve cells

We have long suspected that genetics plays a big role in the etiology of sporadic ALS, which is far more common than the familial form of the disease. However, it was not until the discovery that C9orf72 accounts for a significant portion of sporadic disease that these suspicions were confirmed. This is encouraging news, as it suggests that larger genome-wide association studies (GWAS) will continue to bear fruit in ALS.

1985: The ALS Association funds study of inherited motor neuron disease

TIMELINE 1986: French neurologist Jean-Martin Charcot identifies ALS

bone. Interestingly, OPTN, a major player in the Japanese ALS population, showed up in a genome-wide association study of Paget’s disease.

1968: SOD1 enzyme identified

70s: Programmed cell death in motor neurons demonstrated

1986: Genes for muscular dystrophy identified

1990: Congress declares the 1990s the “Decade of the Brain”

1989: The ALS Association funds search for a common genetic link to ALS

1990: Growth factor CNTF is found to increase survival of motor neurons

No one climbs Everest on their own. Collaboration has become the byword of ALS genetics, as this is the only means by which sufficiently large cohorts of samples can be assembled for genetic research. Numbers alone are not sufficient, however. The samples must be well phenotyped, and there is increasing interest in searching for genetic modifiers that may explain the heterogeneous clinical course of patients. The National Institute for Neurological Disorders and Stroke (NINDS) collection of ALS samples at Coriell, jointly funded by NINDS, the Muscular Dystrophy Association (MDA) and The ALS Association, has become a backbone for ALS genetics. With over 2,000 well phenotyped samples available, the emphasis of this biobank is shifting to collecting familial samples and unaffected parents-affected offspring trios. A spirit of collaboration is also important on the backend. Investigators should be encouraged to make their large genetic datasets publicly available. The National Institutes of Health has established a userfriendly online repository, dbGAP, to do just this. Continued on page 5 The ALS Association begins workshops Glutamate transporter shown to be defective in ALS 1991: Researchers link familial ALS to Chromosome 21

1860 1950 1960 1970 1980 1990 1991

Growth factor BDNF found to increase survival of motor neurons



Call for Research Abstracts

ALS Genetics


Continued from page 4

Investigators around the world can access the raw genetic data, analyze it and combine it with their own data, thereby increasing the power of their studies at no extra cost. Our original genome-wide association study data has been downloaded over a hundred times and incorporated into nearly all subsequent ALS GWASes, highlighting that this resource has long-lasting value to the research community. But what’s next for ALS genetics? We have come a long way, but we have miles further to travel. The ALS research community has published GWAS involving 5,000 cases and 5,000 controls. In contrast, a recent GWAS of myocardial infarction involved 63,000 cases and 130,000 controls, clearly showing that ALS GWAS represents a potential growth industry. Next generation sequencing has already yielded dividends and was instrumental in the discovery of VCP, C9orf72 and PFN1. Additional efforts are underway and will hopefully yield new genes. Another reasons why the C9orf72 discovery caused such a stir in the research community is that it may be amenable to gene therapy targeting the actual gene. Such an approach would have the advantage of being directed at the underlying cause of neurodegeneration, rather than downstream pathways. Preliminary studies applying this technique to SOD1 patients have shown it to be well tolerated. Nevertheless, much

The ALS Association INVESTIGATOR-INITIATED RESEARCH GRANT PROGRAM supports INNOVATIVE research of high scientific merit and relevance to ALS, offering investigator awards in the following categories: Multi-year Grants

Bryan J. Traynor M.B., M.D., Ph.D., M.M.Sc., M.R.C.P.I., Investigator and Chief of the Neuromuscular Diseases, NIH

remains to be done to get such clinical trials up and running for C9orf72 patients. With that in mind, we have established a clinic for C9orf72 patients at the NIH clinical center. Recruiting 60 patients and following them closely over the course of three years, we hope to quantify the natural history of the disease. We are also collecting a range of biospecimens that will be shared with other groups engaged in biomarker development. In this way, we hope to return the knowledge that we have gleaned at the bench back to the bedside to help patients devastated with this fatal illness. I really do believe that the next few years will see us conquer the mountain and have a complete understanding of the genetic architecture underlying ALS. The clouds have just cleared and I can see the summit. C’mon lads, time to break camp, make for Nuptse’s Corner and on to Camp III before the weather changes.

The ALS Association will support research that is projected for periods of up to three (3) years. Funding for multi-year grants is committed for one (1) year only, with noncompetitive renewals conditioned upon results. These applications require strong preliminary data. Awards will be in the amount of up to $80,000 per year. Discovery Grants One-year awards for NEW INVESTIGATORS ENTERING THE FIELD OF ALS. Alternatively, they can be PILOT STUDIES BY ALS INVESTIGATORS. These applications do not require strong preliminary data but must emphasize innovation, scientific merit, feasibility and relevance to ALS. The maximum amount awarded is $40,000. The Milton Safenowitz Post-Doctoral Fellowship for ALS Research Awards The maximum amount awarded is $50,000 per year for two (2) years. Eligibility is limited to those who have been a fellow for one year or less. Request an abstract form for any of these categories from researchgrants@alsa-national. org. See schedule below. GRANT SCHEDULE

Call for Abstracts

Abstracts Due

Full Application Due

Award Announcements

Funding Commences

TIMELINE cont. SOD1 gene mutation (chromosome 21) discovered in familial ALS Trials using glutamate blocker riluzole begin

The ALS Association co-sponsors workshop on high-throughput drug screening with NINDS

Animal studies combining CNTF and BDNF demonstrate decreased motor neuron loss GDNF rescues degenerating motor neurons during development in an in vitro experiment

FDA approves riluzole

Toxic properties of the SOD1 enzyme discovered and linked to familial ALS

RNAi discovered by Craig Mello and Andrew Fire

NINDS issues first ever RFA (request for applications) specifically for ALS research

December 2013 January 17, 2014 March 14, 2014 July 2014 August 1, 2014

A transgenic rat is designed; efforts start on fly model

The ALS Association holds scientific workshop on “Environmental Factors and Genetic Susceptibility”

Attention turns to support cells of nerve tissue to find role in ALS

Aggressive search for new ALS genes funded by The ALS Association

Inflammation and programmed cell death gather research interest

Scientists complete map of mouse genome

ALS2 gene (alsin protein) linked to juvenile ALS

Agency of Toxic Substances and Disease Registries awards five grants focused on ALS

The ALS Association/NINDS collaborative effort begins screening drugs

Department of Defense approves funding for ALS-specific research

1993 1994 1995 1996 1997 1998 1999 2000 2001 2002



Milton Safenowitz Post-Doctoral Fellowships


The ALS Association is especially committed to bringing new concepts and methods into ALS research, and young scientists play an important role in this process. Funding is made possible by the generosity of the Safenowitz family through the Greater New York Chapter of the ALS Association, in memory of Milton, who died of the disease.

Understanding the Relationship Between Progranulin, Programmed Remodeling of the Heat Shock Response Underlies Onset of ALS Sortilin1 and TDP-43 Jennifer Gass, Ph.D. Mayo Clinic, Jacksonville, Florida

Jonathan Labbadia, Ph.D. Northwestern University, Chicago, Illinois

*Funded by Jay & Toshiko Tompkins

* Funded by the Greater Chicago Chapter of The ALS Association through the State of Illinois

A major pathological feature of ALS is the abnormal accumulation of a protein called TDP43. In disease, TDP-43 is redistributed from the nucleus to the cytoplasm of a cell, truncated and abnormally phosphorylated. Mutations in the progranulin gene (GRN) resulting in decreased production of the progranulin protein are associated with TDP-43 pathology in frontotemporal lobar degeneration, a neurodegenerative disease sharing many clinical and pathological features with ALS. Recently, GRN mutations were also discovered in patients with ALS. Within the brain, progranulin levels are regulated by its neuronal receptor, sortilin (SORT1). Currently there is no cure for ALS; however, therapeutic treatments that inhibit the aberrant aggregation, mislocalization and truncation of TDP-43 may prove beneficial for the treatment of ALS. The goal of this project is to determine whether dysfunction of SORT1/progranulin exacerbates TDP-43 pathology and, ultimately, if pharmacological progranulin enhancement alleviates TDP-43 pathology associated with ALS. “I am extremely honored and grateful to be a recipient of the Milton Safenowitz Post-Doctoral Fellowship. Support from this award will allow me to investigate the molecular and cellular mechanisms underlying the development and progression of ALS, specifically elucidating the interactions between TDP-43, sortili and progranulin. Discoveries emerging from my research will provide valuable insight into the processes leading to the accumulation of pathological TDP-43, but also open novel avenues for the development of effective neurotherapeutics for the treatment of ALS.”


The causal relationship between aging and disease remains mysterious; however, an agerelated change in the ability to prevent protein damage has been put forward as a trigger for the onset of neurodegeneration. If DNA is the “blueprint of life,” then the proteins it encodes are its effectors. Cells contain a huge number of proteins that are integral to cell function. If proteins become misfolded or mislocalized they can cause disease, therefore, maintaining protein integrity is essential for health. To achieve this, cells have evolved the protein homeostasis network (PN), a combination of pathways that suppress the presentation and persistence of aberrant proteins. As cells age, the PN becomes compromised, a phenomenon that is exacerbated in many age-related diseases, including ALS. Aging studies in worms have revealed that a “switch-like” re-modeling of the PN is an early event in adulthood. Therefore, Dr. Labbadia and colleagues hypothesize that genetic “re-programming” of the PN underlies the increased susceptibility of individuals to protein misfolding and onset of ALS with age. They will test this hypothesis by identifying the molecular and genetic basis of PN re-modeling and use this knowledge to determine the impact of genetically re-engineering the PN on disease presentation in models of ALS.

Structure and Function of a New Candidate ALS Biomarker RBM45 (RNA-Binding Protein 45) Yang Li, Ph.D. Barrow Neurological Institute, Phoenix, Arizona Members of this lab have recently identified a novel RNA-binding protein, RBM45, implicated in ALS and FTLD. RBM45 was initially identified through its altered levels in the cerebrospinal fluid of ALS patients. Further characterization indicates that RBM45 localizes to cytoplasmic inclusions and co-localizes with TDP-43 and ubiquitin inclusions in ALS and FTLD patients. Dr. Li has been focusing on how RBM45 is involved in the molecular pathway of neurodegeneration. She has discovered the physical interactions between RBM45 and other ALS-associated proteins, and found that RBM45 can self-interact. She will continue to characterize the protein-protein interactions and the self-aggregation propensity of RBM45 in this study. She will also identify the RNA species that are bound by RBM45 by using a novel technique. “I am very honored and grateful to receive the Milton Safenowitz PostDoctoral Fellowship for ALS Research. My research goal is to study a newly identified RNA-binding protein, RBM45 which is implicated in ALS and FTLD. With this encouraging support, I believe the project will provide mechanistic insights into the pathogenesis of ALS and how dysregulation of RNA metabolism leads to neurodegeneration. The novel CLIP-method will also be of help to researchers working on other RNA-binding proteins.” Continued on page 7

“It is a great honor for me to receive a Milton Safenowitz Post-Doctoral Fellowship and I am extremely grateful to The ALS Association for supporting my continued efforts to understand aging and age-related disease. With the support of this award I am confident that I will provide a significant contribution to our understanding of the relationship between aging and ALS.”

Study shows surrounding support cells play key role in ALS Study shows that human embryonic stem cells can be stimulated to produce motor neurons Gulf War study shows that vets deployed to Persian Gulf in 1991 developed ALS at twice the rate of those not deployed there IGF-1 gene therapy study proves beneficial in mice with ALS VEGF gene abnormalities shown to be potential factor in ALS The ALS Association collaborates with U.S. Department of Veterans Affairs to enroll all vets with ALS in registry Early tests of ceftriaxone appear to increase survival in mice with ALS Combination of creatine and minocycline prove more effective together in mouse model than either drug alone

Study implicates smoking as likely risk factor in sporadic ALS Study releases evidence that mitochondrial malfunction may play an important role in ALS Study funded by The ALS Association to find biomarkers in cerebrospinal fluid and blood

Ceftriaxone increases levels of the glutamate transporter GLT1 in a mouse model of ALS First international workshop on frontotemporal dementia discusses link to ALS Stem cells engineered to make GDNF survive when transplanted into rats modeling ALS Early data suggests that mutant SOD1 may be secreted by and may activate microglia Launch of TREAT ALS initiative (Translational Research Advancing Therapies for ALS) to accelerate clinical trials in ALS VEGF increases survival in a rat model of ALS while improving motor performance

ALS patient samples collected to NINDS ALS Repository Repository samples allow genome analysis for sporadic ALS First TREAT ALS clinical trials funded First TREAT ALS clinical trials begun TDP-43 discovered as a common link in FTD, ALS Chromosome 9 region intense focus for FTD

Stem cell study shows SOD1 mutant support cells can kill any motor neuron ALS U.S. registry efforts gaining ground in Congress Fish model of ALS: Progress reported SOD1 in altered form common to both sporadic and inherited ALS Engineered stem cells making GDNF help motor neurons survive in SOD1 mutant rats First genome screening data published based on NINDS ALS Repository

2003 2004 2005 2006 2007


Safenowitz Fellowships Continued from page 6

Use of Extended Irish Kindreds to Identify Novel ALS Variants Russell McLaughlin, Ph.D. Trinity College, Dublin, Ireland * Funded by the Greater Chicago Chapter of The ALS Association through the State of Illinois

Only 10 percent of ALS cases in Ireland can be explained by established genetic factors. This suggests that there are still many ALS genes to be discovered. As technological capabilities increase, whole-genome sequencing represents an exciting avenue in the discovery of novel disease genes. However, careful selection of informative individuals for wholegenome sequencing is paramount in experimental design. The Irish population is well-suited to genetic analysis, given its small size and the relative lack of mixture from other populations. Using existing datasets, Dr. McLaughlin and colleagues have developed methodologies to discover distant relationships among individuals previously assumed unrelated (e.g., third cousins). The proposed project will extend this to identify groups of patients from Ireland and the U.K. who are commonly descended from the same recent ancestor, thus identifying multiple affected members of extended families as ideal candidates for whole-genome sequencing. This is imperative for a better understanding of the underlying mechanism of the disease and will prove vital in patient classification in ALS research, drug development, disease management and clinical trials. “I am extremely excited about this work and am honored that The ALS Association has granted me a Milton Safenowitz Post-Doctoral Fellowship so that I can pursue it further. Through these studies I hope to help to discover some of the unknown causes of ALS and to challenge ideas about the distinction between sporadic and familial forms of the disease. I believe that this will be of great use to other researchers in their future work towards ending this devastating disease.”


Rational Design of Small Molecules Targeting GGGGCC Expanded Repeat in C9orf72 Gene Implicated in ALS-FTD Zhaoming Su, Ph.D. The Scripps Research Institute, La Jolla, California

will search for genetic variants that could contribute to these differences. She plans to investigate 30 well-characterized patients with C9orf72 expansions who have ALS and/or FTD, and compare their pathological and clinical features to their entire genetic background. This comparison will enable her to identify potential disease modifiers, which will be of great predictive value for genetic counselors, and could provide novel targets for the development of treatment strategies.

“This study employs cutting-edge next-generation sequencing techniques to identify disease modifiers in individuals with C9ORF72 repeat expanALS and FTD are incurable neurodegenerasions, the most common known genetic cause of ALS and FTD. My findings tive diseases with a common genetic cause: will, undoubtedly, provide additional insight into C9ORF72-associated an expanded repeat of the sequence GGGGCC in the C9orf72 gene. Like pathobiology. Moreover, they could be a great asset for genetic counselors all genes, C9orf72 is transcribed into RNA, which encodes proteins that complete most of the work in a cell. Once transcribed into RNA, the repeat who would like to offer expansion carriers valuable prognostic information, causes ALS-associated defects. The researchers have previously developed and they might also reveal novel avenues for therapy development, aiming at the postponement of disease symptoms and/or prolongation a strategy to design drugs that target other expanded RNA repeats that of survival.” cause disease. These designed drugs alleviated disease-associated defects in cellular and animal models. They will use similar design principles and strategies to develop drugs that bind the RNA expansion and Determining the Function of C9orf72 Gene and improve ALS-associated defects. “It is my great honor to receive the Milton Safenowitz Post-Doctural Fellowship from The ALS Association. With this support, I will design novel drug-like small molecules targeting the expanded GGGGCC repeat, which has recently become the most common cause of ALS/FTD. These molecules will serve as chemical probes to better understand how RNA contributes to ALS-associated toxicity as well as drugs that lead to the development of novel therapies.

the Pathogenic Mechanisms of its Hexanucleotide Repeat Expansion in ALS

Qiang Zhu, Ph.D. Ludwig Institute For Cancer Research, La Jolla, California

Recently a novel large hexanucleotide repeat expansion in the non-coding region of the Identification of Genetic Disease Modifiers in Patients previously uncharacterized C9orf72 gene has been discovered as the cause of the largest proportion of inherited ALS. Dr. Zhu will exploit gene with C9orf72 Repeat Expansions targeting and BAC transgenic approaches to establish mouse models with Marka van Blitterswijk, M.D., Ph.D. compromised C9orf72 function and/or C9orf72 hexanucleotide expansion Mayo Clinic, Jacksonville, Florida as a means to explore pathogenic mechanisms (including loss of function *Funded by the Greater Chicago Chapter of and/or gain of RNA-mediated toxicity). Moreover, these mice will be used The ALS Association through the State of Illinois for testing efficacy of antisense oligonucleotide for targeted degradation of the C9orf72 mRNA. The discovery of chromosome 9 open reading “I am extremely honored and grateful to receive the Milton Safeframe 72 (C9orf72) repeat expansions as the major genetic cause of ALS and FTD has changed nowitz Post-Doctoral Fellowship for ALS Research. My research focuses on use of genetically modified mice to define pathogenic the field of ALS research. Patients with these repeat expansions can mechanisms in C9orf72 ALS. I am especially excited to examine present with many clinical signs and symptoms, and have demonstrated huge variability in age at onset and disease duration. Dr. van Blitterswijk the efficacy of antisense oligonucleotides that mediate degrada-

Stem cells generated from ALS patients Discovery of DPP6 in two genome-wide association studies in ALS Mutations in TDP-43 linked to familial and sporadic ALS Induced Pluripotent Stem Cell Technology opens up new avenues for ALS

Identification of new gene linked to familial ALS, Fused in Sarcoma (FUS) on Chromosome 16 FDA approval of SOD1 antisense and stem cell trials in U.S.

First patients enrolled for antisense and stem cell trials in U.S.

Ubiquilin-2 discovery; C9orf72 discovery

March: The ALS Association hosts 2nd Drug Discovery Workshop for ALS September: Researchers find genetic region influencing age at which people develop ALS






tion of C9orf72 RNAs carrying repeat expansions as a therapeutic approach in these mice. I believe these in-vivo models will not only be valuable to study disease mechanisms underlying C9orf72 mutation-mediated ALS/FTD, but also provide insight into therapy development for disease treatment.”




JOURNAL NEWS Anti-SOD1 RNA Interference After Onset Extends Survival in ALS Models Peripheral delivery of a single treatment of anti-SOD1 RNA slows progression in a mouse model of ALS even when administered after disease onset, according to a new study. The gene for the RNA was delivered via an adeno-associated virus (AAV) vector. The study suggests that short-term peripheral gene therapy may be therapeutic for some cases of ALS. Intrathecal delivery of AAV9 in macaque monkeys was safe, and high levels of expression of a reporter gene were seen in both neurons and astrocytes throughout the lumbar cord, closest to the injection site. Monkey SOD1 was reduced by 87% in the region. Reporter gene expression was lower but still at 50% or greater throughout the rest of the spinal cord. SOD1 expression was reduced by 60% in the cervical region. Foust KD, Salazar DL, Likhite S, Ferraiuolo L, Ditsworth D, Ilieva H, Meyer K, Schmelzer L, Braun L, Cleveland DW, Kaspar BK. Therapeutic AAV9-mediated Suppression of Mutant SOD1 Slows Disease Progression and Extends Survival in Models of Inherited ALS. Mol Ther. Sep 6, 2013; doi: 10.1038/mt.2013.211. [Epub ahead of print]

A Single Injection of AAV10 Carrying miRNA Targets SOD1 Throughout the Spinal Cord The authors showed that a single injection of the recombinant adenoassociated virus AAV10rh to the lumbar intrathecal space transduced cells along the full length of the spinal cord in mice, with a similar spread in marmosets. In mice, delivery of anti-SOD1 micro-RNA led to RNA interference and reduction of mutant SOD1 expression, leading to a slowing of disease progression. “Our experiments demonstrate that CSF-injection of rAAV can be used to deliver therapeutic genes to treat diseases that afflict broad areas of the CNS and that such treatment is practical and efficacious in a mouse model for ALS,” the authors concluded, and that further development may be warranted for treatment of human disease. Wang H, Yang B, Qiu L, Yang C, Kramer J, Su Q, Guo Y, Brown RH, Gao G, Xu Z. Widespread spinal cord transduction by intrathecal injection of rAAV delivers efficacious RNAi therapy for amyotrophic lateral sclerosis Hum. Mol. Genet. First published online Sep 18, 2013; doi:10.1093/hmg/ddt454

iPS Cells from Sporadic ALS Identify TDP-43 Aggregation Inhibitors The authors developed induced pluripotent stem cell lines from 10 healthy subjects, eight familial and 16 sporadic ALS patients. Three sporadic patient lines displayed aggregation of TDP-43, despite absence of mutation in known ALS genes. TSP-43 aggregations were not seen in cells from healthy subjects or SOD1 ALS cells. In a screen of 1757 bioactive compounds, dose-dependent reduction of aggregation without a change in TDP-43 expression was found for four classes of compounds: cyclin-dependent kinase inhibitors, c-Jun N-terminal kinase inhibitors (JNK), Triptolide, and FDA-approved cardiac glycosides, Digoxin, Lanatoside C and Proscillaridin A. “Our model of human iPSC-derived TDP-43 proteinopathy is amenable to drug discovery and offers promise for discovery of novel disease-modifying therapeutics for ALS and other TDP-43 proteinopathies,” the authors concluded.

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Burkhardt MF, Martinez FJ, Wright S, Ramos C, Volfson D, Mason M, Garnes J, Dang V, Lievers J, Shoukat-Mumtaz U, Martinez R, Gai H, Blake R, Vaisberg E, Grskovic M, Johnson C, Irion S, Bright J, Cooper B, Nguyen L, GriswoldPrenner I, Javaherian A. A cellular model for sporadic ALS using patient-derived induced pluripotent stem cells. Mol Cell Neurosci. Jul 25, 2013; 56C:355-364. doi: 10.1016/j.mcn.2013.07.007. [Epub ahead of print]

ACKNOWLEDGMENT: Richard Robinson, Science Writer

New Insights into FUS Function and Dysfunction FUS (fused in sarcoma) is a 526-amino acid protein with two major RNA-binding domains and a terminal nuclear localization signal. FUS is normally a nuclear protein, but when mutated, it accumulates in cytoplasmic aggregates and leads to ALS, through unknown mechanisms. Several new studies explore the role of both normal and mutant FUS: FUS Aids DNA Repair, and FUS Mutations Impair That Function The FUS protein plays a central role in repair of DNA damage, including double-stranded breaks, according to a new study, and ALS-causing mutations DNA repair. The authors showed that during DNA repair, FUS directly interacts with histone deacetylase 1 (HDAC1), and FUS promotes HDAC1 recruitment to the site of damage. Mutant FUS localized to damaged DNA, but interfered with or was impaired in the formation of DNA repair complexes. Familial ALS patients with FUS mutations had increased levels of DNA damage in motor cortical neurons. “Our current findings indicate that FUS, in conjunction with HDAC1, is important for maintaining genome stability and integrity in the neuron,” the authors concluded. “The impairment of this interaction may contribute to accumulated DNA damage and, eventually, the pathogenesis of ALS.” Wang WY, Pan L, Su SC, Quinn EJ, Sasaki M, Jimenez JC, Mackenzie IR, Huang EJ, Tsai LH. Interaction of FUS and HDAC1 regulates DNA damage response and repair in neurons. Nat Neurosci. Sep 15, 2013; doi: 10.1038/nn.3514.

Aggregation of FUS Can be Mitigated by RNA-dependent Binding into Stress Granules In cultured cells, mislocalized FUS is recruited to stress granules, a site of accumulation of pre-initiation complexes thought to protect cells against stress. Some studies have suggested that recruitment to stress granules is a precondition for FUS aggregation. Here, the authors rebut this claim, showing that removal of the RNA-binding domains in FUS prevent recruitment to stress granules but augment aggregation. In addition, fusing of RNA-binding domains from TDP-43 to the truncated FUS protein restored recruitment to stress granules and reduced aggregation. “Our data indicate that RNA-binding and recruitment to stress granules protect cytoplasmic FUS from aggregation, and loss of this protection may trigger its pathological aggregation in-vivo,” the authors concluded. Shelkovnikova TA, Robinson H, Connor-Robson N, Buchman VL. Recruitment into stress granules prevents irreversible aggregation of FUS protein mislocalized to the cytoplasm. Cell Cycle. Sep 4, 2013;12(19). [Epub ahead of print]

Loss of Nuclear Localization and RNA-Binding Regions Leads to FUS Aggregation The authors expressed FUS lacking both RNA-binding domains and the NLS in neurons of transgenic mice, and found the protein was highly aggregation-prone and triggered neuroinflammation. Motor neurons developed sudden onset of severe damage, with death following within days. “The pattern of pathology in transgenic FUS 1-359 mice recapitulates several key features of human ALS, the authors conclude, with the dynamics of the disease progression compressed in line with shorter mouse lifespan. Our data indicate that neuronal FUS aggregation is sufficient to cause ALS-like phenotype in transgenic mice.” Shelkovnikova TA, Peters OM, Deykin AV, Connor-Robson N, Robinson H, Ustyugov AA, Bachurin SO, Ermolkevich TG, Goldman IL, Sadchikova ER, Kovrazhkina EA, Skvortsova VI, Ling SC, Da Cruz S, Parone PA, Buchman VL, Ninkina NN. Fused in Sarcoma (FUS) Protein Lacking Nuclear Localization Signal (NLS) and Major RNA-Binding Motifs Triggers Proteinopathy and Severe Motor Phenotype in Transgenic Mice. J Biol Chem. Aug 30, 2013; 288(35):25266-74.

Research ALS Today Fall 2013  

The ALS Association's National Research Periodical

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