Annual report 2009
The 3R Research Foundation in 2009
In 2009 the Foundation provided a total of Fr. 634,000 for 17 research projects. The Confederation and Interpharma each made a contribution of Fr. 389,000. The Administrative Board approved 3 new projects, while 6 projects were successfully completed; 18 applications were rejected. The 3R-Info-Bulletins 39-41, which were circulated to around 1,000 readers, included the results of three of the completed projects. The aim of the 3R Internet Training Programme, which was started in 2005, was to provide individual specialised training, but for technical and financial reasons, and because it has not attracted so many users as hoped, it has been decided to abandon this project. The Administrative Board has decided to try to network with other institutions and organisations in the future, in order to achieve greater awareness of the 3R principles.
The 3Rs are Replace, Reduce and Refine animal experimentation. The 3Rs must be the guiding principles behind animal experimentation; if a study can be carried out without using any laboratory animals then such a procedure must be used. If it is essential to use laboratory animals under the terms of animal protection legislation the number used must be kept to a strict minimum. The third “R” requires that animals used for laboratory experiments be made to suffer an absolute minimum of pain and/or stress. The 3R Research Foundation funds research projects whose aim is to improve present-day experimental methods from the point of view of the 3Rs.
Summary of the Year’s Activities
The Foundation’s website
Detailed information about all the Foundation’s activities can be found on its website at www.forschung3r.ch.
17 projects subsidised
A total amount of Fr. 634,308.20 was paid out for 11 ongoing projects and 6 that were completed during 2009.
Three new projects
Three new projects were approved in 2009 for which a total of Fr. 578,500 was earmarked. These new projects are described in detail in the list of funded projects on the Foundation’s website (www.forschung3r.ch/en/projects/index.html).
Engineering of a human brain tumour model to replace animal experimentation (115/09) Dr. Olivier Preynat, Faculty of Medicine, University of Geneva. Brain-like tissue obtained from human stem cells (engineered neural tissue = ENT) will be combined with a glioblastoma-like tumour tissue (engineered glial tumours = EGT) to produce an in vitro model that can be used for examining interactions between human brain and tumour cells as well as for testing potential organ-specific cytostats. The researchers expect to be able to reduce the number of animals required for experiments, which involve considerable suffering.
Organotypic slice cultures derived from brains obtained from slaughterhouses as an in vitro alternative for the investigation of neuroinfectious diseases in ruminants (116/09) Dr. Anna Oevermann and Dr. Torsten Seuberlich, Vetsuisse Faculty, University of Berne. Until now it has only been possible to examine the causes of spongiform encephalitis (e.g. prions) or listeria in ruminants using infected animals. This project involves obtaining tissue samples from various parts of the brain of regularly slaughtered animals (calves and sheep) and culturing them in vitro. If it is possible to maintain structures and functions in the brain tissue samples that are similar to those in vivo it will be possible to better understand the causes (mechanisms) of these diseases as well as to obtain infected tissue for examination without using live animals.
Development of an in vitro model from embryonic stem cells for identifying tissue inflammation as a reaction to implanted material (INFPLANT) (117/09) Prof. Maria Wartenberg, Friedrich-Schiller-University, Jena, and Prof. Heinrich Sauer, Justus-Liebig-University, Giessen, Germany. In the case of artificial implants (teeth, blood vessels, heart valves) it is important to ensure that they are not rejected by the body’s immune system. This issue is currently being studied in animals. In order to replace these animal experiments the research team intends to develop an immune-competent cell culture system from murine embryonic stem cells. Unlike with existing, simple in vitro procedures, the team expect to be able to recognise inflammation and formation of vessels.
Six projects successfully completed
Adjuvanticity of microbial-derived particles and synthetic analogs in vitro (92/04) Prof. Elisabetta Padovan, Gulbenkian Institute of Science, Oeiras, Portugal. Certain adjuvants that stimulate the immune response may also produce toxic side-effects. In order to reduce animal experimentation to test for these unwanted side-effects a three-level cell-culture system using human blood cells (monocytes, dendritic cells and T-cells) has been developed. This system enables researchers to identify possible unwanted toxic characteristics as well as desirable stimulation of the immune system. Consequently, it is possible to largely avoid using laboratory animals through in vivo testing.
Assessment of pain and stress in mice by monitoring gene expression changes (96/05) Dr. Paolo Cinelli, Institute for Laboratory Animal Science, University of Zurich. The aim of this project was to identify pain in animals (rodents) using modified genetic expression. This would provide a basis for developing new methods of recognising pain. Micro-array technology was used to examine two hundred genes while 27 genes were examined using the sensitive RT-PCR (real-time polymerase chain reaction) method. No significant differences were found between the genetic expression in selected areas of the brain of animals following surgical intervention in comparison with earlier.
Establishment of a murine syngeneic co-culture system of intestinal epithelial cells with intraepithelial T-lymphocyte subsets (98/05) Prof. Christoph Müller, Institute of Pathology, University of Berne. A co-culture system of human and murine cells was developed in this project. This enabled the research team to examine the interaction between intestinal epithelial cells and intra-epithelial lymphocytes, which differ from other T-lymphocytes. The results can be used direct for clinical application in humans. This system results in a marked reduction in the number of laboratory animals that will be required for mechanistic testing in the future.
Isolated, autologous blood-perfused heart: Replacement of heterotopic heart transplantation (102/06) Dr. Anna Bogdanova, Institute of Veterinary Physiology, University of Zurich. In this project the researchers successfully developed an ex vivo model of a rat heart that was perfused with the animal’s own blood. This method will enable researchers to carry out ex vivo studies that until now have only been possible using the heterotopic heart transplant method which causes considerable suffering to the laboratory animals used.
Development of in vitro strategies to propagate and characterize hemotrophic mycoplasmas (104/06) Prof. Regina Hofmann-Lehmann, Clinical Laboratory, Vetsuisse Faculty, University of Zurich. The aim of this study was to replace the ethically questionable propagation of hemoplasmas in host animals (e.g. pigs) by an in vitro culture system for M. suis. Using a mycoplasma-specific medium with added fetal calf serum, porcine embryo extract and transferrin the team succeeded in maintaining the continuous growth of M. suis. With this method it is possible to study the characteristics of M. suis without prior propagation in a host animal.
Standardization and pre-validation of MucilAir: A novel in vitro cell model of the human airway epithelium for testing acute and chronic effects of chemical compounds (106/07) Dr. Song Huang, Epithelix Ltd, Plan-les-Ouates. The MucilAir culture system, which consists of a human ciliated lung epithelium, was tested successfully. Procedures were further standardised and one dose-effect ratio was established for each of 9 reference substances (from the EU Acute-Tox Project). The cultured cells showed a stable phenotype and retained their organ-specific characteristics.
3R-Info bulletins are published on the Foundation’s website (www.forschung3r.ch/en/publications/index.html).
Detection of pain in laboratory animals via gene expression? (no. 39, February 2009) In this project, Dr. Paolo Cinelli, University of Zurich attempted to identify pain in animals (rodents) using modified genetic expression. This would provide a basis for developing new methods of recognising pain. Micro-array technology was used to examine two hundred genes while 27 genes were examined using the sensitive RT-PCR (real-time polymerase chain reaction) method. No significant differences were found between the genetic expression in selected areas of the brain of animals following surgical intervention in comparison with earlier.
Refined ex-vivo rodent heart model reduces in vivo experimentation (no. 40, June 2009) Dr. Anna Bogdanova from the University of Zurich and her research team successfully developed an ex vivo model of a rat heart that was perfused with the animal’s own blood. This method will enable researchers to carry out ex vivo studies that until now have only been possible using the heterotopic heart transplant method which causes considerable suffering to the laboratory animals used.
A novel in-vitro cell model of the human airway epithelium (no. 41, October 2009). Dr. Song Huan, Epithelix Ltd, established the optimum culture conditions and defined standard test procedures (duration of exposure, length of testing) for the use of the in vitro lung epithelium model (MucilAir) developed by his company from primary human cells that can be cultured over a period of several months. The tissue showed similar structural and functional characteristics in vivo and makes it possible to elucidate the toxicity of substances and particles that may enter the human airway.
The 3R Internet Training Programme
In 2005 the Foundation set up an internet training programme in German and English at http://3R-training.tierversuch.ch. This programme was intended to provide personal specialised further training. For technical and financial reasons, it is unfortunately not possible to continue to offer the training programme on the platform used so far. Over the past 5 years a total of 81 people have used this internet opportunity to test their knowledge in various fields. To date the programme has not been as successful as hoped among the target market.
1. Origin of the Foundation
The Foundation is a cooperative institution set up by the Parliamentary Group for Animal Experimentation Questions (public organ), Interpharma (association of pharmaceutical companies that carry out research, comprising at present Actelion Ltd, Merck Serono Ltd, Novartis Pharma Ltd, F. Hoffmann-La Roche Ltd, and the associated members Bayer (Switzerland) Ltd, Cilag Ltd and Vifor Ltd) and the Animalfree Research Foundation (animal protection). The Foundation was entered in the commercial register on 18 August, 1987.
The funds provided to subsidise research stem from the Federal Veterinary Office and Interpharma.
2. Purpose of the Foundation
The purpose of the 3R Research Foundation Switzerland is to promote alternative research methods which avoid the use of animals, through grants for research projects. The organisation supports first and foremost projects aimed at developing new methods or refining accepted methods (validation) which offer practical improvements vis-à-vis standard animal experimentation in line with the 3R motto, Replace, Reduce, Refine.
A broad range of projects is sponsored on the condition that they are likely to replace animal experimentation, to reduce the number of animals used or the stress and/or pain suffered. Accordingly, projects based on the Foundation’s three principles and covering any of a broad selection of bio-medical disciplines will be taken into consideration.
3. Organisation of the Foundation
The Administrative Board
The Administrative Board of the Foundation is made up of nine members, three representing the Parliamentary Group for Animal Experimentation Questions (1 seat vacant), two representing animal protection, two from Interpharma and two from the Federal Veterinary Office. Current members are:
Christine Egerszegi-Obrist, member of the National Council, Mellingen (Chairwoman)
The Evaluation Committee
Prof. Peter Maier, University of Zurich (Chairman)
Prof. Peter Maier, University of Zurich
Ernst P. Diener, lawyer, Münsingen
Die Wirtschaftsprüfer.ch AG, Thun
Federal Department of Home Affairs
Articles and statutes of the Foundation
Dr. Martin Reist, Group Head at the Veterinary Public Health Institute at the University of Berne, was elected to the Evaluation Committee for the remaining part of the period of office 2007/2010.
5. Activities during 2009
In its twenty-third year of existence the Administrative Board met three times, namely in May, August and December, for a half-day meeting. Apart from the statutory business concerning the end of the business year 2008, the Board addressed the following issues.
Research funds for 2009 were allotted to 11 projects already underway. In addition, 3 new projects were approved, while 18 applications were rejected. The Board also took note of the final assessment by the Evaluation Committee of 6 projects which had been completed in the previous year. An internal working group was set up to review the scope of the Foundation’s activities. Successfully completed 3R projects are to be monitored more closely by the Foundation in order to encourage the adoption of new methods in practice. On the same principle, more support should be given for verifying newly developed 3R methods. In this connection, the Administrative Board has decided to try to network with other institutions and organisations in the future, in order to achieve greater awareness of the 3R principles.
At its meeting in May, the Board focused on the financial statements for 2008 and the approval of new projects, as well as those projects that had been completed. The question of whether the internet training programme should be continued remained unanswered owing to uncertainty as to its use and necessity. Following the report by the internal working group on the Foundation’s strategic direction, the Board decided to hold a special meeting to discuss this issue.
At the meeting held in August the Board concentrated on the future activities of the Foundation. In the future, apart from its core activity of granting funds for research projects, the 3R Foundation would like to play a more active role as a national platform for spreading awareness of its 3R principles among the Swiss research community. It was decided to ask the members of the Evaluation Committee for their opinion at the next Board meeting.
At the December Board meeting, the focus was on hearing the opinion of the members of the Evaluation Committee, as well as approving new projects. The meeting was rounded off with a dinner. Moreover, the Administrative Board have left the question of the possibility of a new version of the internet training programme open until such time as the level of future interest can be determined.
The Secretary is responsible for the day-to-day running of the Foundation; he deals with all matters that cannot be passed on to anyone else. In particular, he prepares all the necessary information for the Administrative Board to take their decisions, as well as dealing with correspondence with applicants and project managers. The Secretary also deals with payments, book-keeping, closing the books at the end of the financial year and the budget. In addition, he prepares the text of the Annual Report as well as texts for the Foundation’s website. During 2009, a good deal of his load concerned the internal working group on the future scope of the Foundation’s activities.
Under the chairmanship of the Scientific Adviser, the Evaluation Committee held two meetings during the year, where in particular they examined 21 new applications for funding and evaluated the 6 completed projects. The voluntary work of the members of the Evaluation Committee in this connection is much appreciated.
The Scientific Adviser's tasks included publishing the 3R-Info-Bulletins (as a brochure and on the Foundation’s website at www.forschung3r.ch), writing brief scientific reports in English which present the projects receiving funding on the Foundation’s website and regularly updating these reports. As a co-organiser of the EU START-UP project he helped to prepare and ensure the smooth running of the three meetings of experts that took place during the year, as well as taking the minutes that would be included in the final report in 2010. He was also kept busy – as always – advising applicants and project managers, obtaining intermediate reports, evaluating project outlines, dealing with enquiries and explaining why projects had been rejected. Finally, he represented the Foundation at several scientific meetings in Switzerland and abroad, namely as a member of the board of the European Consensus Platform for 3R Alternatives to Animal Experimentation (www.ecopa.eu) in Brussels and as a member of the EPAA Initiative (ec.europa.eu/enterprise/epaa/index_en.htm) also in Brussels.
Overview of the number of applications and approvals
The bar-chart shows that the proportion of applications approved varies only slightly from year to year. The long-term approval rate for applications is around 30%. This figure is dependent on the limited funds available and reflects the great care that is taken to examine applications in the light of their relevance to the 3R principles. Consequently, it often happens that projects that are well structured and of considerable scientific interest are not approved for funding because their relevance to the 3R principles is not sufficiently great. Since the inception of the Foundation, an average of approximately 5 projects have been approved each year.
6. List of new projects approved in 2009
A complete list of projects with summaries of each can be found on the Foundation’s website (www.forschung3r.ch/en/projects/index.html).
The brief scientific project reports in English, which are updated once a year, indicate that almost all projects have progressed well. These reports published on the internet are much appreciated by those involved in the research projects as a platform for presenting their work. From the opposite point of view, this system also enables other researchers all over the world to discover new 3R methods without delay.
In 2009 three more new 3R-Info-Bulletins (ISSN 1421-6590) were published in English and distributed to some 1,000 interested parties. The information bulletins are also published on the Foundation’s website (www.forschung3r.ch/en/publications/index.html), as well as in pdf format.
The latest 3R-Info bulletins are:
8. Financial business
A total of some Fr. 635,551 was paid out for research in 2009 (Fr. 634,308.20 for grants to research projects and Fr. 1,242,85 for participation in conferences). Expenditure on current projects was some Fr. 11,000 over budget (Fr. 623,351.40). On the other hand, relatively little of the Fr. 10,000 budgeted for attendance at conferences was used, so that the overall expenditure for research exceeded budget only by Fr. 2,000.
Operational expenditure for 2009 amounted to Fr. 242,531.85 (project monitoring and information Fr. 128,538.20, administrative costs including office infrastructure Fr. 113,993.65). The total exceeded the budget of Fr. 229,700 by around Fr. 12,831 (5.5%). This was principally due to an invoice for Fr. 17,216 for past maintenance of the 3R Training Programme that had not been budgeted for. Administrative costs were approximately Fr. 2,000 over budget. Total expenditure therefore amounted to Fr. 878,082.90.
On the income side, the equal financial commitment of the federal authorities and Interpharma represented the basic funding for the Foundation’s activities. In 2009 the federal authorities and Interpharma each granted the Foundation Fr. 389,000. As a result of low interest rates, interest on capital was only Fr. 3,949. In addition, income from the 3R Training Course yielded Fr. 1,600, while accumulated contributions for unfavourable age structure from the BVG Insurance Fund resulted in an extraordinary income item of Fr. 7,402.85.
Total income was therefore around Fr. 790,952 while total expenditure amounted to Fr. 878,083, giving an excess of expenditure over income of around Fr. 87,131. The unused contributions item therefore fell from approximately Fr. 559,250 at the end of 2008 to Fr. 472,120 at the end of 2009.
At the end of 2009 the total earmarked for projects approved by the Board but not yet paid out amounted to Fr. 1,092,487.51. This future liability is covered by Interpharma’s new promise of funding. The Foundation’s credit with this institution amounted to Fr. 1,927,000 at the end of 2009.
The budget for 2010 includes around Fr. 685,770 for current projects and a maximum amount of Fr. 500,000 for new projects.
Overview of grants awarded between 1987 and 2009
At the end of 2009 a total of Fr. 16,493,279.15 had been granted for projects and other subsidies, of which Fr. 15,400,791.64 has been paid out so far. Together the federal authorities and Interpharma have contributed Fr. 18,446,000 to the Foundation since 1987.
9. Financial statements
Münsingen, 30 April 2010
3R RESEARCH FOUNDATION
10. Auditors' report to the Administrative Board
Die Wirtschaftsprüfer.ch AG in Thun audited the financial statements for the year according to standards of limited auditing and did not find any indication that the accounts and statements do not correspond to current legislation or the principles and regulations of the Foundation.