Advancing with aptamers What drives scientific change?
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FRAMEâ€™s new Trustee
Non-human primates and drug testing
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Promoting nonanimal laboratory methods through better science
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Issue 74, Spring 2015
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From the Editor
3. Non-human primates and drug testing 4. 3D printing to boost non-animal research 5. Aptamer aid for animal replacement 6. FRAME journal successes 7. News from the FAL 8. Meetings review 9. Giftware Catalogue 11. An end to animal testing? 12. News in brief 13. FRAME training schools 14. New Trustee 15. FRAME annual lecture 16. The FRAME Vision
Welcome to the latest edition of FRAME News. In it you will find updates on FRAMEâ€™s activities and information about developments in alternatives to animal experiments. There are reports of various methods that could replace animal-based models in medical applications. We also describe evidence that old primate-based models are less reliable than was previously thought.
FRAME News 74, Spring 2015 Published by: Fund for the Replacement of Animals in Medical Experiments Russell & Burch House 96-98 North Sherwood Street Nottingham NG1 4EE Registered charity number 259464 www.frame.org.uk Editor: Anne Jeffery E: email@example.com Alternatives to Laboratory Animals (ATLA) www.atla.org.uk Perspectives in Laboratory Animal Science (PiLAS) www.pilas.org.uk 2 | FRAME News 74, Spring 2015
We would like your views on what drives change in science, so inside you will find consideration of the forces that might bring about an end to animal testing, and details of how you can comment on the topic. We also give you the latest on our training schools in experimental design, which help scientists use fewer animals in their work. And there are details of a project being undertaken at the FRAME Alternatives Laboratory. If you would like to contact us about anything here our address and email are detailed below left. Thank you for your continued support in our search for better, more scientifically valid research methods that eliminate the need for laboratory animals. - Anne Jeffery (FRAME News Editor)
Non-human primates and drug testing FRAME is concerned that new experiments to investigate how the liver handles the commonlyused painkiller paracetamol have revealed that a species frequently used to test human drugs cynomolgus monkeys - are not always a reliable model. In fact they are resistant to doses of the drug that would be fatal in humans. The monkeys, also known as crab-eating macaques, are the non-human primates used most commonly to assess drug safety for various reasons, including their size and their relatively close genetic parallels with humans. Prior to their registration all new drugs intended for human use are subjected to a battery of safety studies, including investigations in experimental animals, followed by human clinical trials. In spite of this many hundreds of widely prescribed drugs cause infrequent human adverse reactions (ADRs) occurring at incidences below one in several hundred treated patients. Such human ADRs often affect the liver, although skin and other systems may also be affected. FRAME Scientific Director Dr Gerry Kenna said:
“It is important to develop drug testing strategies which can underpin the selection and development of safer drugs. This new research raises significant concern about the scientific validity to humans of drug safety studies undertaken in primates. “The use of non-human primates in nonclinical safety testing of drug candidates is ethically undesirable and, in view of the substantial cost of such studies, can be expected to increase markedly the cost of drug development – which already is substantial. Such studies should be considered only if they can be shown to be scientifically justifiable and there are not valid alternatives.” More time and money should be invested in cell-based and computer models that would be more reliable. Substantial progress has been made in the development of new methods with the potential to reduce, refine and replace animal studies in the future. The research paper, Metabolism by conjugation appears to confer resistance to paracetamol (acetaminophen) hepatotoxicity in the cynomolgus monkey, is available online at: http://goo.gl/DoOCcY
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3D printing to boost non-animal research The relatively new technology of 3D printing is being adopted by researchers around the world to help human medical advances and to reduce the number of animal experiments being carried out. One of the leading countries in the field is Japan, where the New Energy and Industrial Technology Development Organization (NEDO) has announced a plan to invest ¥2.5 billion (£140 million) over the next five years. Director of NEDO’s biotechnology department Takeshi Sakamoto said the change was being driven partly by the demand for non-animal research in the west. He said criticism of animal testing was growing, particularly in Europe, and he hoped the new methods could help to create tissue models using human cells. Such models will contribute to finding new drugs and developing new treatments because human-based tissues will be more effective, he added. 3D printing is also giving new hope to people needing transplants, Japan Times reported earlier this year. A team at the University of Tokyo School of Medicine is currently working on creating cartilage by using a 3D printer to make replica human body parts using biomaterials called recombinant peptides and various growth factors, then infusing them with human cells. One company in California, USA is already producing test-sized liver tissue samples commercially. Keith Murphy, chairman and chief executive officer of Organovo said:
“Customer response has been strong. Our contracted customers include companies from among the top 25 global pharma companies." There are still many technical problems to overcome, particularly if the system is to be used to produce whole, large organs such as livers. It is difficult to keep cells in the centre of the matrix alive while the rest of the model is still being printed because they are not supplied with oxygen.
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In Australia a team at the University of Sydney has managed to print capillaries, the first stage of vascularisation (incorporating blood vessels) that will remove that hurdle. Research leader Dr Luiz Bertassoni said:
“One of the greatest challenges to the engineering of large tissues and organs is growing a network of blood vessels and capillaries. Replicating the complexity of these networks has been a stumbling block preventing tissue engineering from becoming a real world clinical application. “The possibility of printing threedimensional tissues with functional blood capillaries in the blink of an eye is a game changer. Simplified regenerative materials have long been available, but true regeneration of complex and functional organs is what doctors really want and patients really need, and this is the objective of our work."
Aptamer aid for animal replacement
Aptamers are a new kind of synthetic compound that offer possibilities to replace animals in a range of bioscience applications.FRAME has been investigating the potential of aptamers after hearing from representatives of a manufacturing company at a scientific meeting. Antibodies are produced by white blood cells after any potentially harmful substance or infecting agent – known as an antigen – is recognised by the immune system. They bind to the foreign substance to make it easier for the body to eliminate it. This is called an immune response. As a result, many diagnostic tests use antibodies as a way to identify reactions to substances introduced into the body in experiments. This has many disadvantages, including the fact that it requires living animals – such as rabbits, chickens, mice, rats, guinea pigs - in order to produce the antibodies. Aptamer production does not require animals and can also replace antibodies in many other applications. For example: •antibodies have been used to produce treatments for venomous bites and stings of snakes, spiders or insects •they are also used to diagnose and treat some diseases, such as asthma and chronic obstructive pulmonary disease (COPD) by identifying and targeting substances in the blood that are only found in people with that disease
Dr Arron Tolley, CEO of the York-based, Aptamer Group said: “The failure rate of antibody projects for certain targets is high, leading to multiple attempts when antibodies are not found. In many cases, even when antibodies are found, they are not fit for their intended purpose. Often this is because the target cannot be presented in the same form as in the end test or because the target is not sufficiently reactive or stable enough to elicit an immune response.
“Aptamers however have properties which are fast seeing them become the reagent of choice for those who value speed of delivery, high quality and non-animal testing. Because no animals are used in aptamer selection, molecules that do not elicit an immune response are also good targets for selection purposes”. Arron continued: “Aptamer Group have cutting edge robotic platforms for the identification and characterisation of aptamers against a wide range of targets. Aptamers can be used in all applications where traditional animal-derived antibodies have been used.
“By replacing antibodies with synthetic aptamers we can replace the animal requirement altogether. In addition, the resulting synthetic nucleic acid aptamer will give improved assay consistency, stricter quality control and indefinite supply without ever needing to resort to an animal system”.
• they can also be used in environmental applications for identifying particular pollutants or microorganisms.
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FRAME journal successes FRAME’s scientific journal ATLA has published a number of prestigious papers in recent issues...
At the end of last year ATLA (Alternatives to Laboratory Animals) published a special issue featuring winners in the Lush Prize award scheme. The Lush Prize aims to bring forward the time when safety testing will take place without the use of animals. It focuses on the use of alternatives to animals for toxicity testing of consumer products and ingredients, in a way designed to complement projects that address the use of animals in medical testing. The issue included reports on new models of human bronchial and respiratory systems, how young researchers are contributing to the Three Rs, replacement work taking place in industry, training programmes, and public awareness initiatives. This year ATLA has featured a paper by Dr Tuula Heinonen, winner of the 2014 Björn Ekwall Memorial Award. She was presented with her trophy during the 9th World Congress on Alternatives and Animal Use in the Life Sciences in Prague last year, where she also delivered the memorial lecture: “Better science with human-based organ and tissue models”. Dr Heinonen is from the School of Medicine at the University of Tampere in Finland and the award was presented in recognition of her achievements in promotion and implementation of alternative methods by developing human cell based tissue and organ models. She also established the Finnish Centre for Alternative Methods (FICAM), of which she is Director. She and her group are working to develop human cell based tissue models for biomedical research and toxicology. The first validated model developed by Dr Heinonen’s group is an angiogenesis model applicable for both biomedical and toxicological purposes. Another notable author is Dr Roman Kolar, one of the keynote speakers at the Prague World Congress. Dr Kolar is Deputy Director of the Animal Welfare Academy in Neubiberg, Germany. He has a particular interest in EU policies concerning animal welfare and is a representative on a number of European bodies. He discussed how, in spite of many major political initiatives to reduce the number of animal experiments, totals across Europe continue to rise, and considered some of the hurdles that must be overcome to bring about change. All the articles can be found on the ATLA website, www.atla.org.uk, and many are available free of charge, however, readers will need to register their email address at the site.
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News from the FAL FRAME helps support a laboratory at the University of Nottingham, which investigates new and innovative ways to carry out human medical research without the need for animals. There are currently a number of post-doctoral researchers and PhD students working there on a variety of projects. Among the newest is Tara Stirland, who moved to the FRAME Alternatives Laboratory (FAL) after completing a biology degree at the University’s School of Life Sciences. Tara’s project will involve building three-dimensional skin models from different kinds of cells isolated directly from ethically obtained skin samples. The samples are donated to Nottingham’s City Hospital biobank by patients undergoing mastectomies. Tara will then use the models to investigate how the positive effects that lipids (fatty substances) have on skin health can be enhanced. Completing a PhD was not Tara’s original plan, but after finding she enjoyed writing up her undergraduate dissertation she decided to continue an academic career and was delighted to find an opportunity to work on human tissue.
“I have never supported the use of animals in medical experiments as I not only view it as unethical but acknowledge that the human relevance of obtained results can be questionable. This is especially true for my PhD, as the skin of mice and humans has been shown to differ, both in terms of structure and function. By building a 3D model from human-derived skin cells, I will be able to study skin biology in a more humanrelevant way.” The start of her project has concentrated on building up a deep understanding of her research topic by reading the scientific literature. This was partly driven by the time needed to secure ethically-obtained samples. ^Tara Stirland
“I have never supported the use of animals in medical experiments as I not only view it as unethical but acknowledge that the human relevance of obtained results can be questionable. This is especially true for my PhD, as the skin of mice and humans has been shown to differ, both in terms of structure and function. By building a 3D model from human-derived skin cells, I will be able to study skin biology in a more human-relevant way.”
“As my project is very different from what I was doing before, it has been helpful to be able to spend time studying the literature before undertaking any practical work. I now feel that I am in a position where I understand the background surrounding my PhD and therefore I have been able to plan my work in a more targeted fashion.” While the project is still at its early stages Tara has her sights set on her future. “After my PhD I wish to continue in a career in which I can make a meaningful contribution towards scientific knowledge. I am fascinated by the complexity of the human body and how pharmaceuticals can be designed to alter its biology and thus improve health."
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Meetings Throughout the year FRAME staff attend a variety of meetings held by other organisations in order to share news and information across the scientific community. At some we offer advice, at others we learn about new methods and techniques or changes to legislation governing the use of animals in laboratories. Here are highlights from some we have attended. RSPCA Lay Members Forum All establishments breeding or using animals for scientific procedures in the UK must have an Animal Welfare and Ethical Review Body (AWERB). The RSPCA runs regular meetings for those involved in AWERBs to provide them with helpful information and advice. FRAME attends the meetings to give its own perspective. Among the topics at the latest meeting was the question of increased use of fish in experiments. Some researchers see fish as an alternative to higher animals, but there is increasing evidence that fish can experience pain and suffering so there is a need for serious consideration of the ethics of using them. The meeting also heard Dr Gillian Currie from the University of Edinburgh discuss the importance of good experimental design in order to minimise the number of animals used in experiments. In particular she concentrated on proper randomisation and blind testing in order to avoid unintentional bias in the results. Proper experimental design is one of FRAME's key messages. We promote it through regular training schools around Europe and Scandinavia. See page 13 for more information.
The Boyd Group The Boyd Group is a UK based forum for open exchange of views on issues related to the use of animals in science. Members span a range of expertise and perspectives and the group aims to promote dialogue and, where there is consensus, to recommend practical steps towards achieving common goals. FRAME attended a meeting on openness in animal research and testing. Participants were divided into small groups with a mixture of backgrounds who discussed and identified points of agreement or disagreement. They then considered how progress might be made where disagreement exists. Topics included: What should â€˜opennessâ€™ look like? How well do current initiatives meet these expectations? What are the challenges to delivering openness?
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The group also discussed how openness initiatives might be improved. There was a lot of focus on getting the public involved but it was recognised that increased openness might not lead directly to public involvement and that science practices might need to change. FRAME Scientific Programme Manager Michelle HudsonShore, who attended the Group, said: "This meeting was useful. It highlighted that there is a lot of consensus regarding openness but that there are some barriers that will need to be overcome. It also showed that the concordat on openness is not enough to drive the aspirational openness discussed. Openness is still seen by many animal users and/or institutions as risky, so there needs to be an improvement in mutual trust. That is, scientists trusting the public and other interested parties, and the public and others trusting scientists. Openness is important for FRAME as it allows us to access data and to generate a dialogue with scientists."
Imperial College London Animal Research Forum Following severe criticism of management of its animal research facility last year, Imperial College London opened its doors to interested parties to show the improvements it has made in response. FRAME attended the open day. Visitors were able to visit labs and were given examples of new protocols that have been put in place. Imperial has also set up a detailed governance structure to ensure new policies are operated fully. The college now plans to stage similar events annually as part of its improvements, and its commitment to openness about the work it carries out and the conditions under which its research animals are held.
FRIENDS OF FRAME giftware Friends of FRAME is the group for FRAME supporters who help us through regular donations and by spreading the word about FRAME’s work. They are individuals who believe in what we stand for and the way we go about promoting the end of animal experiments. We could not continue our vital work without the support of people like them and we are always grateful for their help.
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What will drive the end of animal testing? In spite of campaigns by many bodies and individuals for an end to animal experimentation the numbers used in procedures in UK laboratories continue to rise. If a change is to happen there must be a driver behind it. Here we discuss some of the forces involved in the question of implementing Replacement.
Public opinion In any democratic system, the voice of the public should be heard and acted upon. Historically, the power of the people to initiate change has been demonstrated. Campaigns against animal testing on cosmetics began in the early 1970s and continued until a full ban was introduced across Europe in 2013. The use of animals for scientific experimentation is a polarising topic, but if enough of the populace feels that change is required, and is willing to act on it, that could be the strongest driving force.
Government legislation The capacity for control, whether to encourage or hinder social, economic, or industrial change, is central to the power of a government. Through legislative authority a government has the final say regarding how a country will respond to a given situation. In the EU, the clearest example would be the passing of Directive 2010/63/EU which regulates the use and protection of animals used for scientific purposes. Evidently there is some level of effectiveness related to the decision-making abilities of the legislative body.
Institutions and organisations (such as industry, academia, charities, etc.) Ultimately, neither the government nor the general public are the actual bodies conducting the research that uses animals. Those on the front line, including universities, are the ones responsible for the scientific use of animals. While working within all legal boundaries, each institute or laboratory can set its own precedent or criteria for the application and frequency of its animal use. As opposed to relying entirely on the regulators and general opinion, individuals working directly with animals can make the most important changes effectively and promptly but they are often driven along established lines through habit or resistance to unproven ideas.
We would welcome your views. A survey can be found on the FRAME website that will enable you to comment.
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news in brief Home Office latest on household product testing The Home Office has announced that there will be a ban on testing household products on animals with effect from October. It will apply to products bought by the public for use at home, including cleaning products, polishes, detergents, paint and decorating materials, laundry products and adhesives. It does not cover pesticides, medical products, or products intended for use by professionals in an industrial setting. FRAME’s Scientific Director Dr Gerry Kenna said: “FRAME is delighted to see this change, which could mean a huge reduction in the number of animals used for safety testing each year in the UK. It is heartening that the Minister stressed the Government’s determination to apply strict controls on any tests that will still be permitted under the new rules.” Earlier this year Derby North MP Chris Wiliamson asked a question in the House of Commons about what research the Home Office had commissioned into the contribution to the UK economy that such tests made. Mr Williamson told FRAME: "I have campaigned on animal rights and welfare since the mid-1970s. I'm a trustee of the League Against Cruel Sports and have been a vegan since 1976. I am against unnecessary experiments on animals." Home Office Minister Lynne Featherstone said: "Although superficially straightforward, this issue has not been easy to resolve. We cannot ban testing which may be required under EU law.
"We also need to be cautious that we do not develop a solution that precludes research that is essential or drives necessary research overseas."
^ Kevin with his poster in Prague
Kevin takes on a permanent role After completing a one-year contract with FRAME, Kevin Coll has accepted a permanent post as a Scientific Officer. During his first year Kevin worked on a number of projects, including the new FRAME Alternatives Timeline, as well as attending several conferences, and presenting a poster about the increase in use of genetically modified mice at the 9th World Congress on Alternatives and Animal Use in the Life Sciences in Prague. Kevin returned to his native Canada over Christmas to spend time with his family but came back to England early in January to start his new job. He is now looking forward to working on projects to promote the Three Rs and promote FRAME and its work. 12 | FRAME News 74, Spring 2015
Training scientists to use fewer animals Although FRAMEâ€™s ultimate vision is a world where the default position in biosciences is to use non-animal methods it acknowledges that animal experiments do take place, and campaigns to reduce the numbers involved. In their 1959 book The Principles of Humane Experimental Technique, William Russell and Rex Burch identified that the way in which reduction in laboratory animal use can most effectively be achieved is by rigorous experimental design and appropriate statistical analysis of any results. To help promote those ideals FRAME runs regular training schools in experimental design and statistics. The schools were launched in 2008 and at least one a year has been held since then, in various venues across the UK, Europe and Scandinavia. The latest took place at the end of March at the University of Coimbra in Portugal. The schools aim to give participants an appreciation of what they need to consider when designing a biomedical experiment and to provide them with the basic skills needed to plan and analyse their work in such a way as to minimise the number of animals needed and maximise quality and relevance of scientific output. This is in line with the Three Rs principle set out by Russell and Burch: Replacement, Reduction and Refinement. Where replacement of animal models is not possible the numbers used should be reduced to the lowest necessary to give meaningful results and any suffering minimised. The main objective of the schools is to reduce the number of animals used in research, education and testing without compromising the quality of data or hindering scientific progress. They aim to provide researchers with an understanding of basic experimental design concepts that they do not seem to be gaining from other sources. A key outcome is increased awareness and understanding among scientists about the need to reduce animal numbers in experiments and to refine procedures undertaken on them which will lead to a fall in the number of laboratory animals used and a reduction in the suffering they encounter. In addition, with a better understanding of how to properly design and effectively analyse their experimental programmes, participants will go on to produce higher quality science, which has made most efficient use of resultant improved data.
^ The University of Coimbra, Portugal
Delegates report that the schools are extremely helpful. One said: â€œThis course gave an excellent presentation of what is often a poorly-taught or often overlooked aspect of research training. It finally explained all the issues surrounding experimental design and actually linked biological relevance, practical consideration, ethics and the Three Rs to planned statistical tests. I have no doubt that I will draw on what I have learned in this course in the future.â€? FRAME News 74, Spring 2015 | 13
new trustee appointed FRAME has appointed a new Trustee, Gary Thomson. Born in Whitby and educated in St Andrews in Scotland, Gary went to the University of Stirling to study biochemistry, but dropped out in the third year to become a professional musician after a band he was in gained a recording contract with Virgin Records. He spent the next 10 years playing trumpet and keyboards in various bands, during which time he settled in Nottingham with his wife Anna. When their son was born he set up an internet design and IT business so he could be at home more often. In 2003 he sold the business to go to art college to study painting. He now works with a film and TV production company and is currently engaged in producing his first feature film. He has long-standing experience of charity working. His parents founded Worldwide Cancer Research and, as children, he and his siblings were involved in fundraising and promoting activities for it.
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“At that time much of cancer research was conducted using animals, but thanks to the work that FRAME and others are doing that continues to decrease. Like most people I hate the idea that animals are so casually used but many people are only here because of the animal work done in the past, so I accept it is sometimes necessary. “I am sure that new technologies and techniques will eventually lead to no animals being used at all. Till then we must practice the Three Rs. That is why I believe FRAME is more important than ever. I have known several of the FRAME Trustees for more than 20 years and was delighted to be asked to help.” Gary is married to FRAME’s chair of Trustees Dr Anna Cadogan and they have two children Alex (17) and Alice (15). They also have three pet pigs and two cats.
FRAME Annual Lecture The latest FRAME Annual Lecture was given by Dr Malcolm Skingle of GlaxoSmithKline. He manages academic liaison at GSK with staff in the US and UK. His role involves working with several groups outside the company, including Government departments, research and funding councils. He has worked in the pharmaceutical industry his entire career, was formerly a neuropharmacologist and has over 60 publications including articles on the interface between industry and academia. He sits on many external bodies including several UK university department advisory groups. He chairs several groups including the Diamond (Synchotron) Industrial Advisory Board, and the ABPI academic liaison group. He has also served on the NC3Rs Board and chaired the NC3Rs Studentship Panel. Dr Skingle was awarded a CBE in the 2009 Queen’s Birthday Honours List in recognition of his contribution to the pharmaceutical industry. He also holds an Honorary Professorship from the College of Medical and Dental Sciences at the University of Birmingham, honorary degrees from the University of Hertfordshire and the University of Dundee, and a fellowship from Imperial College London medical faculty. The talk detailed GSK’s commitment to the Three Rs agenda and his work in collaborating with external groups to deliver research of high quality. FRAME was delighted to welcome long-time supporter Dr Brian May among the lecture audience.
A recording is available to watch online at www.youtube.com — search for 'FRAME Annual Lecture'. FRAME News 74, Spring 2015 | 15
FRAME's vision FRAMEâ€™s vision is a world where nonanimal methods are accepted as scientific best practice. Its mission is to enable and support the timely development and use of scientifically valid methods that provide reliable data, and replace the need for animal experiments.
Why are non-animal methods needed? Animal studies cannot be ethically justified unless they will provide valuable biomedical data that have the potential to enhance human or animal health substantially and there is no alternative method. For example, if they are intended to provide novel insight into disease processes, to enable discovery or development of new pharmaceuticals to treat human or animal diseases, or to support human or animal risk assessment of chemicals to which they are exposed. In many cases it is also difficult to justify them on scientific grounds. Laboratory animal strains are not representative of the diverse human population. We do not have good animal models of many human diseases or adverse reactions caused by potential new drugs. Animal studies are expensive, slow and unsuitable in many cases, such as drug discovery phases where there is abundant chemical choice, or testing safety of large numbers of household products, industrial or agricultural chemicals.
Why does FRAME stand in the middle ground? We are often asked why we do not take a stronger stance in either direction in the debate over animal experiments. There are already advocates at the extremes but that has so far failed to bring about any major steps forward in the search for working non-animal methods. Laboratory animals exist and scientists will continue to use them until better, more effective, more scientifically valid methods are found. Developing and implementing effective non-animal methods is feasible, but difficult. Wasting time and energy on arguing about the current situation gains nothing. By taking the middle ground FRAME is able to talk to people from all viewpoints and find common ground where advances might be made. The goal is to share views, engage and foster a way forwards, not 'win or lose' and create enemies. Achieving full replacement of animals will take time and effort and will require co-operation. It will be achieved in part from in vivo studies in animals (undertaken according to Three Rs principles), in part from in vivo studies in humans and in part through mathematical data modelling. We must all work in partnership, not in conflict, if we are to move forwards and find effective, scientifically valid, non-animal models.
Published on Apr 1, 2015
Twice-yearly newsletter of the Fund for the Replacement of Animals in Medical Experiments. Promoting non-animal laboratory methods through b...