Emerging technologies and the impact on the ethical use of animals in scienceEmerging technologies and the impact on the ethical use of animals in science

Allison Guy*, MSc, Gilly Griffin*, PhD, & Clément Gauthier*, PhD*Canadian Council on Animal Care • 1510-130 Albert, Ottawa ON, Canada K1P 5G4 • www.ccac.ca

AbstractThe Canadian Council on Animal Care (CCAC) is the national organization responsiblefor overseeing the care and use of animals in science. Like most other national oversightsystems, whether legislated or not, the CCAC system is based on the Russell and Burchtenet of the Three Rs (Replacement, Reduction and Refinement). Publication of the mouseand human genome sequences and the ability to manipulate an animal's genome has ledto a rapid increase in the numbers of animals being "designed" to answer questions con-cerning human disease. There are underlying ethical and animal welfare challenges asso-ciated with the use of these animals, including the large numbers of animals that are usedin the pre-experimental production phase, as well as the potential pain and distress asso-ciated with the procedures used and with the disease models themselves. The CCAC isin the process of developing guidelines on: genetically-engineered animals, in particular toaddress these ethical issues and to improve the welfare of the animals.

The development of high-throughput genomics, proteomics and metabolomics technolo-gies are resulting in simultaneous monitoring of the expression of large numbers of indi-vidual genes and proteins, permitting a more detailed study of disease processes. It maybe too early to say what effect this will have on the overall pattern of animal use in sci-ence. However, the potential of genomics technology as a Three Rs tool has already beenrecognized. The emergence of these technologies and the elucidation of biological mech-anisms they provide may allow the identification of opportunities to replace animal usewith in vitro alternatives. In addition, the ability to examine the sub-clinical effects ofsmaller quantities of chemicals holds the promise to minimize pain and distress and toreduce the numbers of animals involved as the examination of earlier and multiple exper-imental endpoints within one animal becomes possible. The challenge will be to ensurethat these emerging technologies are harnessed appropriately to support sound science,including the ethical use of animals.

The CCACThe CCAC is the national peer review organization responsible for overseeing the careand use of animals involved in research, teaching and testing throughout Canada. TheCCAC comprises 25 member organizations whose representatives include veterinarians,scientists, educators, and delegates from industry and the animal welfare movement.Through programs of guidelines, assessment, and education, implemented with the helpof volunteers, including the members of animal care committees at institutions across thenation, the CCAC ensures that the use of animals for research, teaching and testingemploys optimal physical and psychological care without compromising scientificintegrity. The Guidelines Program achieves this goal through the development of guide-lines that assist both investigators and animal care committee members to implementappropriate standards for housing, husbandry and procedures at the local level. Theseguidelines are also used by the Assessment Program in carrying out quality assuranceevaluations of institutional animal care and use programs, and form the basis of their rec-ommendations for the improvement of animal care and use at an institution. The infor-mation gained through these assessments is further used to expand on or refine existingguidelines as well as to create new ones. The CCAC also acts to promote an increasedlevel of knowledge, awareness and sensitivity to relevant ethical principles through theirEducation, Training and Communications Program.

The Ethical Use of Animals in ScienceThere is considerable debate about the rights and wrongs of animal use in science. Somepeople argue that the protection of animal welfare should be paramount, while othersbelieve that the moral arguments support continued use of animals in scientific research. Ingeneral, societal opinion weighs in favour of animal research provided it is conductedunder stringent controls1,2 and in particular, Canadians feel it is acceptable to conductresearch using animals provided the animals are subjected to minimal pain and theresearch substantially benefits humans or animals3. The principles developed by Englishphysiologist and physician, Marshall Hall4, to describe acceptable use of animals in researchin 1847, still hold true today for the ethical treatment of animals in science and form thebasis for the CCAC policy statement on: the ethics of animal investigation†5.

The CCAC and the Three RsIn 1954, the University Federation for Animal Welfare appointed two English scientists,William Russell and Rex Burch, to conduct a study of the ethical treatment of laboratory ani-mals. This led to the publication of The Principles of Humane Experimental Technique in 1959,which outlines the principles of a universal ethic for animals used in research6. These prin-ciples are still relevant today and are commonly referred to as the Three Rs. The tenet of theThree Rs is now incorporated in legislation regulating the use of animals for scientific pur-poses in several countries. In Canada, where there can be no federal legislation in this areabecause of the constitutional division of power7, the CCAC as the national quasi-regulatorybody has incorporated this tenet into its fundamental policy document5.

Application of the Three Rs in GenomicsPublication of the mouse8,9 and human10 genome sequences and the ability to manipulatean animal's genome has led to a rapid increase in the numbers of animals being designedto answer questions concerning human disease, basic biological mechanisms and theresponse to toxic and pharmacological compounds (Figure 1). There are underlying ethi-cal and animal welfare challenges associated with the use of these animals, including thelarge numbers of animals that are used in the development of a genetically-engineeredstrain, as well as the potential pain and distress associated with the procedures used andwith the disease models themselves3. Furthermore, these new technologies raise questionsabout 'intrinsic factors' that urgently need to be answered, such as determining "to whatextent human ends and purposes should be permitted to override the essential nature ofa particular species"11. The CCAC is in the process of developing guidelines on: genetically-engineered animals, a revision of CCAC guidelines on: transgenic animals12, in particular toaddress these ethical issues and to improve the welfare of the animals. Genome Canadaalso acknowledges the far reaching implications of emerging genomic technology and hasbegun to fund research projects that focus on the ethical, environmental, economic, legaland social issues related to genomics research (GE3LS). Animal welfare has been identi-fied as a societal value that has the potential to be compromised by the new applicationsof biotechnology. Therefore, interface with the CCAC was recommended to the SteeringCommittee of Genome Canada at the GE3LS Workshop as part of successful outcomes forimproving integration of GE3LS into genomic science13.

National surveillance bodies will need to make a concerted effort toaddress current inefficiencies in the creation of genetically modifiedanimals if an escalation in numbers is to be avoided.11

Gauthier & Griffin, 2005

Marshall Hall's PrMarshall Hall's Pr inciplesinciples4

1. No experiment should take place if the necessary information could be gained byobservation.

2. Only experiments that would result in the fulfillment of clearly defined and attain-able aims ought to proceed.

3. Unnecessary repetition of an experiment must be avoided – particularly if rep-utable physiologists had been responsible for the original experiment.

4. All experiments must be conducted with a minimum of suffering.

5. All physiological experiments should be witnessed by peers, further reducing theneed for repetition.

Acknowledgment

The authors wish to thank Emily Verlinden for her considerable assistance in the preparation of this poster.

† The use of animals in research, teaching, and testing is acceptable only if it promises to con-tribute to understanding of fundamental biological principles, or to the development ofknowledge that can reasonably be expected to benefit humans or animals.

The The Three RsThree Rs

Replacement: Animals may be used only if the researcher's best efforts to find areplacement by which to obtain the required information have failed.

Reduction: The fewest animals appropriate to provide valid information and sta-tistical significance should be used.

Refinement: The most humane, least invasive techniques must be used.

Figure 1: Procedures involving normal, mutant and genetically modified animals in the UK from 1995-200514.

The use of normal animals in scientific procedures has declined from 2.27 in 1995 to 1.65 million proceduresin 2005, while the use of genetically-modified animals has quadrupled since 1995 and now represents 33%of all scientific procedures (up from 8% in 1995). Meanwhile, the use of mutant animals, those animals withnaturally occurring harmful genetic defects, has only increased from 8% of all procedures to 10%. The UKdata is shown above as animal use data collected by the CCAC does not differentiate between genetically-normal and genetically-engineered animals. This trend was also shown by Gauthier & Griffin (2005).

1 Market & Opinion Research International (MORI) (2000)Animals in Medicine and Science. General Public ResearchConducted for Medical Research Council. London, UK: MORI.

2 Canadian Public Health Association (2001) Animal-to-humantransplantation: Should Canada proceed? 25pp. Ottawa, ON:Canadian Public Health Association.

3 Canadian Biotechnology Advisory Committee (CBAC) (2001)Biotechnology and Intellectual Property: Patenting of Higher LifeForms and Related Issues. Interim Report to the Government ofCanada Biotechnology Ministerial Coordinating Committee.54pp. Ottawa, ON: CBAC.

4 Hall M. (1847) On experiments in physiology, as a question ofmedical ethics. The Lancet 49(1220):58-60.

5 Canadian Council on Animal Care (CCAC) (1989) CCAC policystatement on: ethics of animal investigation. Ottawa, ON: CCAC.Available at: http://www.ccac.ca/en/CCAC_Programs/Guidelines_Policies/POLICIES/policy.htm

6 Russell W.M.S. & Burch R.L. (1959) Principles of HumaneExperimental Technique. London: Metheun. 38pp. UniversitiesFederation for Animal Welfare (UFAW), Potters Bar, Herts,UK: England. Special edition (1992).

7 Wilson P. (1998) Legislative Jurisdiction Over Animals Used inResearch, Teaching and Testing. Canadian Council on AnimalCare commissioned legal opinion. Ottawa, ON: Osler, Hoskin& Harcourt.

8 Gregory S.G. et al. (2002) A physical map of the mousegenome. Nature 418:743-750.

9 Waterston R.H. et al. (2002) Mouse Genome SequencingConsortium. Initial sequencing and comparative analysis ofthe mouse genome. Nature 420:520-562.

10 International Human Genome Sequencing Consortium (2001)Initial sequencing and analysis of the human genome. Nature409:860-921.

11 Gauthier C. & Griffin G. (2005) Using animals in research,teaching and testing. Animal Welfare: global issues, trends andchallenges. OiE Scientific and Technical Review 24(2):735-745.

12 Canadian Council on Animal Care (CCAC) (1997) CCAC guide-lines on: transgenic animals. Ottawa, ON: CCAC. Available at:http://www.ccac.ca/en/CCAC_Programs/Guidelines_Policies/POLICIES/policy.htm

13 Genome Canada (2006) Improving integration of GE3LS intoGenomic Science. GE3LS Workshop Report. Ottawa, ON:Genome Canada.

14 Secretary of State for the Home Department (2006) Statistics ofScientific Procedures on Living Animals. London, UK: HomeOffice.

15 Thole E. (2002) Genomics & Alternatives To Animal Use.Maastricht, the Netherlands: Netherlands GenomicsInitiative.

16 Canadian Council on Animal Care (CCAC) (1998) CCAC guide-lines on: choosing an appropriate endpoint in experiments using ani-mals for research, teaching and testing. Ottawa, ON: CCAC.Available at: http://www.ccac.ca/en/CCAC_Programs/Guidelines_Policies/POLICIES/policy.htm

References

Although there are concerns that emerging genomic technologies may lead to an initialincrease in animal use in research, these technologies have the potential to decrease thenumber of animals used in the long term. The development of high-throughputgenomics, proteomics and metabolomics technologies enable simultaneous monitoring ofthe expression of large numbers of individual genes and proteins, permitting a moredetailed study of disease processes and toxicity mechanisms. These emerging technolo-gies should allow for a better understanding of basic biological mechanisms whichshould allow researchers to ask more specific questions and refine their experimental pro-cedures. It may be too early to say what effect this will have on the overall pattern of ani-mal use in science. However, the potential of genomics technology as a Three Rs tool hasalready been recognized. The following is a list of possible ways emerging genomic tech-nologies may contribute to the implementation of the Three Rs based on the recommen-dations arising from a conference on Genomics & Alternatives to Animal Use15 organized byNetherlands Genomics Initiative and the Netherlands Centre Alternatives to Animal Usein 2004.

Replacement:

•• Elucidation of biological mechanisms, as well as correlations between animal, invitro, and human data, should increase the validity of in vitro studies.

•• The use of human cells or stem cells can increase the physiological relevance of invitro methods to the biological processes in humans.

•• Human studies could be used earlier in pharmacological or toxicity testing as geneactivity can be measured at the sub-clinical level or using low doses.

Reduction:

•• Genomic technologies enable the simultaneous measurement of different param-eters which allows for the collection of more data from each animal model.

•• Establishing a template of the characteristics of known toxicants should enable thepre-screening of compounds and should lead to a decrease in the number of ani-mal studies required for the categorization of chemicals.

Refinement:

•• Identification of biomarkers, such as changes in protein levels, and gene expres-sion patterns induced by a certain compound or treatment can lead to earlier(more appropriate) endpoints. This is in line with CCAC guidelines on: choosing anappropriate endpoint in experiments using animals for research, teaching and testing16.

•• Microarray technology has increased the sensitivity of detection methods andshould allow effects of a compound to be seen at lower doses.

•• Insertion of human genes into animal models may enable better extrapolationfrom these models to humans.

•• Using gene-expression, a short term assay, to predict long-term effects candecrease the pain and distress of animal models.

•• Assessing the homology between species allows the selection of the most appro-priate animal model for the study.