Interviews

Antidote Europe (AE): Could you tell our readers how your team came to launch this company and how you became interested in non animal replacement methods?

Samuel Constant (SC): There were two reasons: (1). A desire to develop and promote model systems to predict toxicity that were scientifically and ethically more acceptable than using laboratory animals, and (2) the challenge of starting our own company.

In the course of our research work in academia, we realised that many modern in vitro models were not readily accessible to researchers. Not being able to make use of these modern methods in research can make it difficult to reproduce data published in the scientific literature.

In view of these challenges, and thanks to the efforts of our colleague Ludovic Wiszniewski in the field of cystic fibrosis, we decided to set up the company Epithelix in order to develop and market in vitro test models using tissue and primary cells sourced from the human respiratory system.

AE : Could you describe your unique product « MucilAir » bearing in mind that most of our readers do not have a science background?

SC : Epithelix has developed a unique product, a human reconstituted tissue, which  can be maintained in culture for extended periods of time. The product is called “MucilAir” and is made of respiratory epithelial cells obtained from surgical waste. This model is a kind of “mini-lung” containing the same cells and performing the same functions as the upper respiratory tract in humans. MucilAir mimics the first line of defence of the respiratory system against the outside environment. Its unique long shelf life of one year means that it can be used for both acute and chronic studies. MucilAir is therefore very versatile and useful for studying (1) the toxic effects of environmental pollutants, the effect of e-cigarettes, chemicals and other inhaled substances, etc. and (2) the efficacy of new treatments for respiratory diseases, especially asthma, chronic obstructive bronchitis, cystic fibrosis as well as bacterial and viral infections.

AE : It would appear that there are very few in vitro methods available to replace the use of animals in inhalation tests for pharmaceutical and chemical products. What is the status of your product “MucilAir” with respect to the replacement of animals by industry?

SC : There are sadly no in vitro methods currently approved by the official body of the European Union (EURL ECVAM) to replace the use of animals for inhalation toxicity assessment. As a result, the pharmaceutical, cosmetics, and chemical industries continue to use animals in order to market their products. The aim of Epithelix is to change this. We are currently developing new tests, based on MucilAir, in order to study the short-term and long-term effect of inhaled substances on human health.

Each test method is different, depending on whether the aim is to test corrosivity, irritancy, sensitisation and even carcinogenicity of inhaled products. Currently, the most advanced test model is trans-epithelial permeability testing, which we are developing in collaboration with the team of Prof Carrupt at the University of Geneva and that of Dr Sandra Coecke at EURL ECVAM. This test method provides toxicokinetic data [the rate at which a chemical will enter the body and what happens to it once it is in the body] and measures the amount of inhaled substance that crosses the epithelium and can potentially be found in the blood stream. We are working hard to get this test method approved as quickly as possible.

AE : What in your view are the biggest obstacles to replacing the use of animals for your products? How can progress be made?

SC : The foremost challenge for our company is to maintain our independence in order to preserve our values. For a company like Epithelix, whose strategy is to re-invest its profits into research and development, each day is a struggle. It is clear that smaller companies find it much harder to finance their projects.

Another challenge is time. The time required to validate a new method [by the regulatory authorities] is beyond our control.

Yet another challenge is conservatism. It is much riskier for a regulator or an industrial toxicologist to endorse a new in vitro method than a “tried and trusted” traditional animal test.

Finally, it should be noted that the cost of validating a new non animal method falls largely on the shoulders of the company developing the method. In some cases, the cost is simply prohibitive. It would therefore be helpful to establish a fund whose aim would be to financially support the validation of alternative methods.

AE : We would like to thank you sincerely for giving up the time to take part in this  interview. Are there any additional thoughts that you would like to share with our readers?

SC : I would like to thank you too. The European Citizen’s Initiative [www.stopvivisection.eu] has been a huge success and I congratulate Antidote for its part in this. This initiative is very encouraging for researchers like us, who want to promote alternative methods. We would also like to sincerely thank the Swiss League Against Vivisection, the Egon Naef Foundation and the committee of Proanima, for their support.

Short CV

Dr Samuel Constant is co-founder and COO of Epithelix (http://www.epithelix.com), a Swiss biotech company that specialises in tissue engineering. Epithelix is a leader in the field of in vitro efficacy and toxicity testing of human respiratory passages. Epithelix developed a unique 3D in vitro model of the respiratory mucosa and associated test systems for the study of human pathologies such as asthma, cystic fibrosis and chronic obstructive bronchitis. Samuel is responsible for management and development strategies for the company and also for collaborations with other private and public research groups.

Over the past six years, Dr Samuel Constant and his team have focused their research efforts on the development of new in vitro approaches to the study of the effects of xenobiotics on the human respiratory system. His efforts have provided a model system for assessing repeat dose toxicity (90 day exposure) as well as assessing respiratory absorption and metabolism, inflammation, and identifying respiratory sensitisers. In the years since 2006, Dr Samuel Constant and his team have been the recipients of 14 prizes in recognition of their scientific achievements, technological innovation and financial development.

Antidote Europe (AE): At what point in your science training did you begin to question the value of animal models with respect to human medicine? What are your current views on animal experiments in the field of research and testing and how did you come about them?

Meg Lewis (ML):

It wasn’t until my current post that I became aware of the inefficacies of animal experiments, almost 4 years after I’d completed my PhD training.

The accepted thinking had always been that animal models are used where it would be unethical to do a test on a human – for example to find out if a compound is toxic. The motivation behind animal experiments is that they are supposed to give us information about whether something would be effective in humans, whether it would cause harm and to what extent. The underlying assumption is that animals are good models of humans, and that the information obtained from experimenting on them allows us to make accurate predictions of what happens in humans. I was shocked to learn the extent to which this is simply not the case.

We’ve known about the inefficacies of animal experiments for a long while, certainly since the 1960s, so I’m surprised that it hasn’t cropped up during my education. The figures are staggering – 94% of the drugs that make it past pre-clinical testing in animals fail at the clinical trials stage because they are either toxic or not effective in humans. There were an estimated 197,000 deaths related to adverse drug reactions in the EU in 2011. The increasing cost of getting a single drug to market (now an estimated average in the range of $1.5bn – $6bn), and the diminishing returns the pharmaceutical industry is seeing are bringing the sector into economic crisis. Animals are not humans, and respond differently to drugs. We need to accept these insufficiencies in current testing and move away from the use of animals for research and toxicology by developing in vitro tests and assays based on better science, that are more sensitive and more accurate than animal tests.

The replacement of animals in research and testing is not going to happen overnight, and may not even happen completely, but the earlier we start, and the more time and money we invest in alternative solutions, the more successful the end result will be.

AE: Today you are employed full-time as R&D Manager in a forward-looking biotech company (Kirkstall Ltd). Could you describe to our readers your proprietary product “Quasi-Vivo System” and what role it plays in the replacement of animals in research and testing?

ML:

Kirkstall has a strong commitment to the 3Rs. The Quasi-Vivo system is a tool for cell culture, and can be thought of as a “Lego kit” for cell biologists, allowing them to culture cells in an environment which more closely represents the way that cells grow in the human body.

In order to successfully replicate the in vivo environment in vitro, a number of key in vivo aspects are required: the architecture of the organ and secreted factors that would be present in the extracellular matrix, tissue movement and plasticity, blood flow, specific oxygen pressures, and co-culture of the different cell types that make up an organ. The balance of these elements will depend on the requirements of each tissue, for example, bone cells will function best under higher pressures on a rigid 3D scaffold, and lung cells will prefer an environment under lower pressure with an air interface.

The central element of the Quasi-Vivo system is to expose cells to flow, with different chambers designed to support specific tissues in specific ways, by simulating their environment either through the chamber itself (as in our air-liquid interface chamber designed for skin or lung culture) or through compatibility with other technologies that replicate other key elements of the in vivo environment. Most standard cell culture is done in cell culture medium under static conditions. The introduction of a dynamic environment, where the medium is circulated by a pump and passes across cells, ensures cells are constantly provided with fresh nutrients, stimulated by low shear stress, and kept oxygenated. Recirculation of medium allows cells to condition their environment with secreted cytokines, as they would do in the body, increasing their longevity and proliferation, and enabling different tissues to talk to each other between chambers modelling inter-organ communication in vivo. This type of system makes it possible to look at how the drug metabolic products affect different organs without using an animal model.

Building a representative in vitro environment encourages cells to behave as they do in the body – we have had great success with our system in getting  cells to function in similar ways to in vivo in terms of gene expression, drug metabolism, and improved longevity in culture. The system has been used to detect the toxicity of the drugs diclofenac, roflumilast and cyclophosphamide with between 4 and 30x greater sensitivity than standard in vitro assays.

AE: What advantages does your “Quasi-Vivo System” have over those of your competitors? Is there scope for improvement of your product or collaboration with like-minded scientists?

ML:

Groups developing technology in this area of bioscience are often doing so out of a drive to improve in vitro cell culture, and make it more representative so that data from in vitro experiments gives a more accurate picture of what is happening in the human in vivo environment. This has led to ingenious solutions for reducing and eventually replacing the need for animals in research, including spheroids to mimic 3D tissue strictures, and the ‘human on a chip’ idea of fluidic microchips where small populations of cells can communicate with each other through microscopic channels, in addition to our own system.

The Quasi-Vivo system is designed around the novel idea of individual chambers that can be interconnected to model the in vivo environment communications between organs in an easy-to-use way. The system enables researchers to study these communications and the effects they have on populations of cells without having to worry about technical problems such as leaks (which often plague other flow systems), bubbles and pressure artefacts (problems that come with microfluidics). The dimensions of the chambers are modelled on a standard 24 well plate commonly used for in vitro research, so they are compatible with 3D cell scaffolds designed for that size of culture vessel. Cells can be easily recovered after experiments, in numbers suitable for analysis of gene expression or protein levels.

We welcome any collaborations with like-minded scientists as we continue to develop our system for new applications. Our system is compatible with many other alternative technologies, such as induced pluripotent stem cells and 3D culture scaffolds. We believe that by working together with these technology providers we get the best out of the cells, and stand the best chance of making realistic in vitro models to replace animal use.

AE: How do you see the future of R&D in your field? If you had a budget of £ 10 million today, what would be your number one R&D priority?

ML:

The future is very bright, not only in the field of alternative technologies, where recognition and funding are increasing year on year, but also in the fields these technologies impact, such as general bioscience research, toxicology testing and so on. We know what the problems are, and we know the solutions, so it is just a matter of connecting the dots, developing these alternative technologies in synergy and optimising them to provide the most realistic models possible.

My research priorities for alternative technologies over the next 5-10 years would be to continue to develop and optimise new models and explore new applications, particularly where animal use is prevalent as in cancer research and toxicology testing. Alternative models will also be able to support the growing sphere of personalised medicine, which is reliant on individual human data, for example by providing a platform for testing a patient’s susceptibility to a drug for multiple organs in a connected system, using cells derived from a patient’s blood and then re-differentiated to lung, kidney, liver or heart. It will be very interesting to watch how the landscape develops.

AE: We thank you for giving up your time for this interview. Are there any additional comments that you would like to add that were not covered in the interview?

ML:

In my opinion, the long-term answer to widespread adoption of alternative technologies lies in education and in streamlining the alternative methods validation process.

If students could be taught about the pros and cons of animal testing and the availability of other solutions, they would come out of school or University with an informed, critical perspective on animal experiments, and this would encourage them to question and consider alternatives rather than re-enforcing the doctrine that animal experiments are necessary in order for developments in medicine and science to continue. Part of our aim as a company is to teach scientists the value of alternative technologies, and how to adopt them, in our monthly training courses.

Validation remains a significant roadblock to uptake of alternative methods by industry. At 10 years and €5m average investment to get a single assay validated by ECVAM, the cost is simply too great for most small companies, but without it, it is very difficult to convince contract research organisations and pharmaceuticals to use new technology and change their approaches (though not impossible).

CV:

Meg Lewis graduated from the University of Sheffield with a BSc in genetics and microbiology and a PhD in molecular biochemistry, after which she worked for the Department for Business, Innovation and Skills before joining Kirkstall in May 2011. As Principal Researcher, she oversaw the completion of the second TSB-funded Grant for Research and Development project, and had involvement in a number of commercial and collaborative projects.

In her current role as Research and Development Manager, she co-ordinates the research programme and research strategy, puts together new collaborative projects with leading research groups from across the world, and this year established a course programme training researchers how to use the Quasi-Vivo system.

In addition to research work, she is involved in supporting commercial relationships and academic partnerships in a technical and supervisory role, and in 2012 founded the Kirkstall Young Scientist Programme for science graduates in collaboration with the Job Centre to help new researchers into jobs in relevant fields.

Antidote Europe (AE): Your academic background is that of physics. Please could you tell our readers how you made the transition from this field to becoming the head of what is surely the most successful organisation for promoting the replacement of harmful animal use in education and training worldwide?  Was there a particular event in your life that altered the course of your career or was this a gradual process?

Nick Jukes (NJ): My interests had always been in the arts and the sciences, but I chose the sciences at university. My previously passive support of environmental awareness and animal rights flourished towards the end of my degree due to the vibrant atmosphere of debate and campaigning action. In the late 1980’s I took on an active leadership role within the local radical green group, addressing a wide range of issues from anti-vivisection to anarchism, health to human rights.

A science education can in theory encourage critical thinking, but it often teaches conformity and an avoidance of the concept of social responsibility. I realised that my real education was that provided by grassroots political activism and a community of campaigners. For the 1990’s my work and life was a combination of peaceful direct action, information networking, permaculture design and the promotion of alternatives to harmful animal use.

The humane education and alternatives work was done through EuroNICHE, whose first public conference I had attended in 1989. As the network I was now leading grew more international and was renamed InterNICHE, I appreciated the potential of the organisation and movement to empower people and to create sustainable radical change.

While the InterNICHE focus is very specific and appears to have a narrow application, in fact we are informed by an understanding of intersectionality: how different forms of discrimination and exploitation are interlinked, and how a holistic approach can help bring multiple positive impact that reaches across education, the professions and society as a whole. This is clearly illustrated by the intersection of student rights to a high quality, ethical education on the one hand, and animal rights and freedoms on the other. We articulate a bold vision of humane education and full replacement, and employ a creative, solution-oriented approach to make the vision real. Despite its challenges, the work of helping to catalyse many small revolutions in people’s psychology and practice across the world is truly rewarding.

AE: Could you give our readers an overview of your work? Do you mainly focus at university education level or other areas as well? What countries have you visited in your campaigning?

NJ: We work primarily at university level, addressing the use of animals and alternatives in practical classes for biology, medicine and veterinary medicine. Harmful animal use includes animal experimentation and the dissection of purpose-killed animals in disciplines such as anatomy, physiology, pharmacology, clinical skills and surgery. We also address professional training, such as in advanced surgery training courses, and sometimes secondary level education, where dissection might still be performed.

Alternatives include non-animal alternative tools such as digital video, multimedia software and virtual reality (VR), training models, mannekins and simulators. Alternatives also include approaches such as student self-experimentation, the use of ethically sourced animal cadavers, and the clinical learning opportunities of work on animal patients. Our own experience of the pedagogical, ethical and economic advantages of alternatives over harmful animal use is backed up by that of teachers, students and many published studies. Indeed, these innovative and humane methods are now widely available and often no longer considered ‘alternative’, but the norm.

InterNICHE works internationally with teachers, students and campaigners to facilitate the implementation of alternatives and the replacement of harmful animal use. InterNICHE projects include information resources such as the book from Guinea Pig to Computer Mouse and the on-line Studies Database and Alternatives Database (the latter with details of over 1000 alternatives); further multi-language website resources; Alternative Loan Systems, or libraries, to enable the trial, assessment and demonstration of alternatives; a Humane Education Award grant program to support local development and implementation of alternatives; empowerment of student conscientious objectors; a range of freeware and other low cost alternatives; and conferences, exhibitions, outreach visits and training worldwide.

I have travelled very widely – so far having visited nearly 50 countries. My large carbon footprint is my one big compromise in a life in which I aspire to compromise very little. Nevertheless, I always try to maximise the impact of my visits. By being part of a large network of people who want to take action in this area, there has been the opportunity to work in countries as diverse as Brazil, South Africa, Iran, Russia, Uzbekistan, India and China, as well as across Europe.

AE: Our director, Andre Menache, was very privileged to travel with you on a lecture tour of South African universities in August 2012, which included medical and veterinary schools. Could you share with us some of the highlights of that tour in terms of the sort of positive feedback you received from the teaching staff?

NJ: The outreach to South Africa was very successful. Over a week of events on replacement alternatives in education, research and testing were held, comprising the first major alternatives outreach in the country. The main workshops were followed by seminars at universities at 6 cities across the country. The education and training workshop explored best practice and the advantages of humane learning tools, illustrated with video clips and demonstrations. A Multimedia Exhibition provided access to nearly 100 learning tools from across the disciplines, including models, mannekins and simulators, and software that had been networked across 50 computers.

A live demonstration of laparoscopic surgery illustrated the potential of advanced training tools for superior surgical skills acquisition and the replacement of animal experiments. Ethically sourced organs were perfused using the POP (Pulsating Organ Perfusion) Trainer. The organs derived from a companion animal dog that had been euthanised for medical reasons, following the strict InterNICHE Policy on replacement methods.

For the research and testing focus, presentations were given by experts Barbara Grune from the German Federal Institute for Risk Assessment (BfR) and Andre Menache. Andre explored the use of animal models and human-based research in an age of personalised medicine. He reflected on ethical committees and their effectiveness, with a call to establish a level playing field, and questioned the use of sentient animals in fundamental research, suggesting it is time to raise the bar.

Barbara Grune introduced alternatives in these fields and provided a review of legislative developments concerning animal experimentation, such as the new EU Directive 2010/63/EU. She described the work of AXLR8 and its efforts to accelerate the transition to a toxicity pathway-based paradigm for chemical safety assessment. Finally she reviewed funding and validation of alternatives, and reviewed global information resources and techniques for reliable searches on alternatives.

The workshops had an audience of over 70 people who comprised a diverse cross-section of representatives from academic, industry and regulatory bodies. The range and high level of the questions were very promising for the future. And as has been reported from previous InterNICHE outreach, comments were received from participants who had arrived cynical but left convinced of the need for replacement – and willing to help achieve it.

AE: Based on your extensive travelling and your encounters with various academic institutions, what would you say are the remaining obstacles to the wider use and implementation of non harmful teaching and training methods?

 NJ: All the obstacles to full replacement are surmountable. InterNICHE and others are working to remove real and perceived obstacles and to make the transition easy and desirable. The projects and resources we offer have been developed for that purpose. Amongst other advantages, alternatives are usually cheaper than the costs associated with animal experiments, so widespread appreciation of this will help in the transition. For some countries, access to computer hardware can sometimes hinder implementation, but just as the reach of technology is increasing, so is the price decreasing.

Rigid adherence to tradition is often a problem, but when this inertia is recognised as blocking the enhancement of education and training – as well as being unscientific – then the benefits of developments in technology and the sharing of excellent new teaching tools can be reaped. There are also a number of myths and misconceptions that need naming and shaming:

First is that full replacement is not realistic, and that the 3Rs of reduction, replacement and refinement are appropriate in education and training. The reality is that full replacement has already been achieved in countless universities across the world – to everyone’s benefit – and is a far better approach than any other that holds back the implementation of advanced methods.

Second is that alternatives are not ‘the real thing’, and that animal experiments are. In fact, animal experimentation is a method, and should not be confused with an objective. The ‘real thing’ should refer to the range of teaching objectives and the process of successfully meeting them. A focus on ethical and effective acquisition of knowledge, skills and attitudes, rather than emotional attachment to a specific method of questionable benefit, is essential.

Third is that if there are no more animal experiments then students and trainees will have no learning and training tools at all, and no access to animals. The reality is that alternatives significantly add to and improve education and training. Replacement means replacement of harmful animal use, so as well as introducing non-animal methods for all students and trainees, those like vets who really needs hands-on experience of animals can be exposed to neutral or beneficial work with animals, for example within the healing context of a clinic.

AE: We would like to thank you very much for giving up the time to take part in this interview, knowing what a very busy schedule you have. In conclusion, are there any additional thoughts or comments that you would like to share that were not covered in the interview?

NJ: There’s a growing momentum for replacement of harmful animal use by alternatives in education and training, so now is a very exciting time. Formal agreements that we have signed with Russian and Ukrainian universities have led to the annual saving of over 50,000 animals. In India, the national academic council for zoology has come out against dissection, so as alternatives are introduced, up to 19 million animals will no longer be killed each year.

One reason for the success is the fact that alternatives provide a win-win solution. Students win, because they have access to best practice learning tools and a good learning environment; teachers win because they can more effectively share their knowledge and skills; society and the professions win because there is a new generation of better trained, critical thinking, ethically literate and compassionate graduates; and of course the animals win: no more killing, and no other forms of harm.

Biography: Nick Jukes

Nick Jukes graduated in the physical sciences from the University of Leicester, England, in 1988. Since the mid-1990s he has co-ordinated InterNICHE, helping to build a diverse network of individuals and campaigns working for curricular transformation and humane education within the life sciences. He co-produced the award-winning film Alternatives in Education (InterNICHE, 1999), and project managed and co-authored the 2^nd edition of from Guinea Pig to Computer Mouse (InterNICHE, 2003). These have been distributed widely across the world in many translations. He has helped establish a range of other resources and projects on alternatives, including the InterNICHE Alternatives Loan Systems and Humane Education Awards (from 2002). He co-organised the Alternatives, Animal Welfare and the Curriculum series of training seminars (India 2004), the 2^nd InterNICHE Conference Alternatives in the Mainstream: Innovations in life science education and training (Norway 2005), and the Multimedia Exhibitions at the 5^th, 7^th and 8^th World Congresses on Alternatives and Animal Use in the Life Sciences (Germany 2005, Italy 2009 and Canada 2011). He also co-organised the Latin American series of seminars Alternative Methods for an Ethical Education: Best Practice and Innovation in the Life Sciences (2008) as well as the 1^st Pan-African Seminar and 1^st North Africa and Middle East Seminar on Alternatives to Animal Experiments in Education and Training (Kenya 2009 and Egypt 2010). Recent (2012) outreach has included seminars and exhibitions across Central Asia and Latin America. InterNICHE activity has led to widespread direct replacement of animal experiments and dissections. Nick is currently doing further global outreach, producing a new film on alternatives in veterinary education and training, and co-organising the 3^rd InterNICHE Conference (England 2013). He is also interested in photography and psychosynthesis.