Internal consistency and compatibility of the 3Rs and 3Vs principles for project evaluation of animal research.

Using animals for research raises ethical concerns that are addressed in project evaluation by weighing expected harm to animals against expected benefit to society. A harm-benefit analysis (HBA) relies on two preconditions: (a) the study protocol is scientifically suitable and (b) the use of (sentient) animals and harm imposed on them are necessary for achieving the study's aims. The 3Rs (Replace, Reduce and Refine) provide a guiding principle for evaluating whether the use of animals, their number and the harm imposed on them are necessary. A similar guiding principle for evaluating whether a study protocol is scientifically suitable has recently been proposed: the 3Vs principle referring to the three main aspects of scientific validity in animal research (construct, internal and external validity). Here, we analyse the internal consistency and compatibility of these two principles, address conflicts within and between the 3Rs and 3Vs principles and discuss their implications for project evaluation. We show that a few conflicts and trade-offs exist, but that these can be resolved either by appropriate study designs or by ethical deliberation in the HBA. In combination, the 3Vs, 3Rs and the HBA thus offer a coherent framework for a logically structured evaluation procedure to decide about the legitimacy of animal research projects.

From Mice to Monkeys? Beyond Orthodox Approaches to the Ethics of Animal Model Choice.

Recent developments in genome editing tools, along with limits in the translational potential of rodent models of human disease, have spurred renewed biomedical research interest in large mammals like nonhuman primates, pigs, and dogs. Such scientific developments raise ethical issues about the use of these animals in comparison with smaller mammals, such as mice and rats. To examine these ethical questions, we first consider standard (or "orthodox") approaches, including ethics oversight within biomedical research communities, and critical theoretical reflections on animal research, including rights-based and utilitarian approaches. We argue that oversight of biomedical research offers guidance on the profession's permitted uses of animals within a research setting and orthodox approaches to animal ethics questions when and whether animals should be used in biomedicine; however, neither approach sufficiently investigates the nuances of ethical practices within the research setting. To fill this lacuna, we consider a virtue ethical approach to the use of specific animal models in biomedicine. From this perspective, we argued that limitations on flourishing for large mammals in a research setting, as well as potential human-animal bonds, are two sources of likely ethical tensions in animal care and use in the context of larger mammals.

Reevaluating Benefits in the Moral Justification of Animal Research: A Comment on "Necessary Conditions for Morally Responsible Animal Research".

In a recent paper in Cambridge Quarterly of Healthcare Ethics on the necessary conditions for morally responsible animal research David DeGrazia and Jeff Sebo claim that the key requirements for morally responsible animal research are (1) an assertion of sufficient net benefit, (2) a worthwhile-life condition, and (3) a no-unnecessary-harm condition. With regards to the assertion (or expectation) of sufficient net benefit (ASNB), the authors claim that morally responsible research offers unique benefits to humans that outweigh the costs and harms to humans and animals. In this commentary we will raise epistemic, practical, and ethical challenges to DeGrazia and Sebo's emphasis on benefits in the prospective assessment of research studies involving animals. We do not disagree with DeGrazia and Sebo that, at the theoretical level, the benefits of research justify our using animals. Our contribution intends to clarify, at the practical level, how we should understand benefits in the prospective assessment and moral justification of animal research. We argue that ASNB should be understood as an assessment of Expectation of Knowledge Production (EKP) in the prospective assessment and justification of animal research. EKP breaks down into two further claims: (1) that morally responsible research generates knowledge worth having and (2) that morally responsible research is designed and executed to produce generalizable knowledge. We understand the condition called knowledge worth having as scientists' testing a hypothesis that, whether verified or falsified, advances an important interest, and production of generalizable knowledge in terms of scientific integrity. Generalizable knowledge refers to experimental results that generalize to a larger population beyond the animals studied. Generalizable scientific knowledge is reliable, replicable, and accurately descriptive. In sum, morally responsible research will be designed and carefully executed to successfully test a hypothesis that, whether verified or falsified, advances important interests. Our formulation of EKP, crucially, does not require further showing that an experiment involving animals will produce societal benefits.

Is selecting better than modifying? An investigation of arguments against germline gene editing as compared to preimplantation genetic diagnosis.

BACKGROUND: Recent scientific advances in the field of gene editing have led to a renewed discussion on the moral acceptability of human germline modifications. Gene editing methods can be used on human embryos and gametes in order to change DNA sequences that are associated with diseases. Modifying the human germline, however, is currently illegal in many countries but has been suggested as a 'last resort' option in some reports. In contrast, preimplantation genetic (PGD) diagnosis is now a well-established practice within reproductive medicine. Both methods can be used to prevent children from being born with severe genetic diseases. MAIN TEXT: This paper focuses on four moral concerns raised in the debate about germline gene editing (GGE) and applies them to the practice of PGD for comparison: Violation of human dignity, disrespect of the autonomy and the physical integrity of the future child, discrimination of people living with a disability and the fear of slippery slope towards immoral usage of the technology, e.g. designing children for specific third party interests. Our analysis did not reveal any fundamental differences with regard to the four concerns. CONCLUSION: We argue that with regard to the four arguments analyzed in this paper germline gene editing should be considered morally (at least) as acceptable as the selection of genomes on the basis of PGD. However, we also argue that any application of GGE in reproductive medicine should be put on hold until thorough and comprehensive laws have been implemented to prevent the abuse of GGE for non-medical enhancement.

No Pain, No Gain? In Defence of Genetically Disenhancing (Most) Research Animals.

Every year, around 12 million animals are used for the purpose of scientific research in the European Union alone. The procedures performed on them often cause significant pain and suffering. Despite regulations aimed at reducing this suffering, we can expect millions of research animals to continue to suffer in the near to mid-term future. Given this reality, we propose the use of gene editing to create research animals with a reduced capacity for suffering, in particular, from pain. We argue that our proposal would be in line with moral principles embedded in European regulations regarding animal research, and that it would facilitate compliance with these regulations. We also respond to the strongest argument against our proposal-the 'no pain no gain' argument.

Necessary, but Not Sufficient. The Benefit Concept in the Project Evaluation of Animal Research in the Context of Directive 2010/63/EU.

Directive 2010/63/EU (henceforth "Directive") on the protection of animals used for scientific purposes mandates that every project proposal in EU member states involving procedures on living non-human vertebrates and cephalopods has to be approved in an review process, including a harm-benefit-analysis (HBA), to assess "whether the harm to the animals in terms of suffering, pain and distress is justified by the expected outcome taking into account ethical consideration and may ultimately benefit human beings, animals or the environment". Despite the justifying relevance of "outcome" and "benefit", it remains unclear how to understand these concepts. However, national authorities and applicants require a clear understanding of this to carry out a HBA. To analyze the underlying premises of the HBA and its consequences for the evaluation process, we introduce a heuristic to analyze the relation between "outcome", "benefit" and "prospective benefit assessment". We then apply the heuristic to all seven legitimate purposes for animal research stated in the Directive, namely basic research, translational or applied research, product safety, education and training, protection of the environment, preservation of species and forensic inquiries. As we show, regardless of which purpose is aimed for, applicants are hard-pressed to demonstrate tangible benefits in a prospective assessment. In the HBA, this becomes a problem since-as we argue-the only reasonable, expected and tangible outcome of research can ever be knowledge. The potential long-term benefits on the basis of gained knowledge are unforeseeable and impossible to predict. Research is bound to fall short of these proclaimed societal benefits and its credibility will suffer as long as research has to validate itself through short-term societal benefit. We propose to revise the ethical evaluation based on the HBA and we think it necessary to develop an alternative model for project evaluation that focuses on the value of knowledge as a scientific outcome as a necessary but not sufficient condition for societal benefit.

The Road to Hell Is Paved with Good Intentions: Why Harm-Benefit Analysis and Its Emphasis on Practical Benefit Jeopardizes the Credibility of Research.

It is our concern that European Union Directive 2010/63/EU with its current project evaluation of animal research in the form of a harm-benefit analysis may lead to an erosion of the credibility of research. The HBA assesses whether the inflicted harm on animals is outweighed by potential prospective benefits. Recent literature on prospective benefit analysis prioritizes "societal benefits" that have a foreseeable, positive impact on humans, animals, or the environment over benefit in the form of knowledge. In this study, we will argue that whether practical benefits are realized is (a) impossible to predict and (b) exceeds the scope and responsibility of researchers. Furthermore, we believe that the emphasis on practical benefits has the drawback of driving researchers into speculation on the societal benefit of their research and, therefore, into promising too much, thereby leading to a loss of trust and credibility. Thus, the concepts of benefit and benefit assessment in the HBA require a re-evaluation in a spirit that embraces the value of knowledge in our society. The generation of scientific knowledge has been utilised to great benefit for humans, animals, and the environment. The HBA, as it currently stands, tends to turn this idea upside down and implies that research is of value only if the resulting findings bring about immediate societal benefit.

The microtubule stabilizer patupilone counteracts ionizing radiation-induced matrix metalloproteinase activity and tumor cell invasion.

BACKGROUND: Ionizing radiation (IR) in combination with microtubule stabilizing agents (MSA) is a promising combined treatment modality. Supra-additive treatment responses might result from direct tumor cell killing and cooperative indirect, tumor cell-mediated effects on the tumor microenvironment. Here we investigated deregulation of matrix metalloproteinase (MMP) activity, as an important component of the tumor microenvironment, by the combined treatment modality of IR with the clinically relevant MSA patupilone. METHODS: Expression, secretion and activity of MMPs and related tissue inhibitors of metalloproteinases (TIMPs) were determined in cell extracts and conditioned media derived from human fibrosarcoma HT1080 and human glioblastoma U251 tumor cells in response to treatment with IR and the MSA patupilone. Treatment-dependent changes of the invasive capacities of these tumor cell lines were analysed using a Transwell invasion assay. Control experiments were performed using TIMP-directed siRNA and TIMP-directed inhibitory antibodies. RESULTS: Enzymatic activity of secreted MMPs was determined after treatment with patupilone and irradiation in the human fibrosarcoma HT1080 and the human glioblastoma U251 tumor cell line. IR enhanced the activity of secreted MMPs up to 2-fold and cellular pretreatment with low dose patupilone (0.05-0.2 nM) counteracted specifically the IR-induced MMP activity. The cell invasive capacity of HT1080 and U251 cells was increased after irradiation with 2 Gy by 30% and 50%, respectively, and patupilone treatment completely abrogated IR-induced cell invasion. Patupilone did not alter the level of MMP expression, but interestingly, the protein level of secreted TIMP-1 and TIMP-2 was lower after combined treatment than after irradiation treatment alone. Furthermore, siRNA depletion of TIMP-1 or TIMP-2 prevented IR-mediated induction of MMP activity and cell invasion. CONCLUSIONS: These results indicate that patupilone counteracts an IR-induced MMP activation process by the reduction of secreted TIMP-1 and TIMP-2 proteins, which are required for activation of MMPs. Since IR-induced MMP activity could contribute to tumor progression, treatment combination of IR with patupilone might be of great clinical benefit for tumor therapy.