The course programme is split into specialty topics as outlined below, with Day 2 consisting of two tracks from which attendees can choose.
AAALAC International updates, AALAS and FELASA working groups, education, harm/benefit analysis, and harmonisation of training requirements
Directive 2010/63/EU – Progress to Date and Future Challenges
Speaker: Susanna Louhimies, Policy Co-ordinator, European Commission, Belgium
Harm-Benefit Analysis in Light of the AALAS/FELASA Working Group
Speaker: Aurora Brønstad, University of Bergen, Norway (Part 1 +2)
Harmonisation and Mutual Recognition of Education and Training in Europe
Speaker: Patri Vergara, Universitat Autònoma de Barcelona, Spain
Although the European Directive EU63 2010 made only general provision on the education and training (E & T) requirements for different personnel working with experimental animals, several organizations have been working to get mutual recognition of E & T in laboratory animal science, thereby facilitating the mobility of laboratory animal science (LAS) personnel in Europe.
Firstly, the European Commission with the support of all Member States have established an Expert Working Group (EWG). This group has produced a Guidance document including the principles and criteria for a modular, outcome-based education and training framework.
Secondly, the FELASA Accreditation system for LAS programmes, implemented in 2003, has contributed to the harmonization of LAS training programs, particularly for personnel conducting and designing experiments with animals. FELASA has adapted its requirements in accordance with the EWG Guidance document and has accredited new courses under this system. FELASA is still the only pan-European accreditation system.
Thirdly, in anticipating the difficulty of harmonising programmes and obtaining recognition of competences by each Member State, the EWG has proposed the Education and Training Platform for Laboratory Animal Science (ETPLAS). Until now, ETPLAS has focused on two documents relevant for mutual recognition: Minimal Requirements for Mutual Recognition of Courses in Laboratory Animal Science and Accreditation bodies, Quality assurance of LAS Education and Training to promote the development of quality systems in LAS E & T. ETPLAS also provides information about courses available in Europe.
Finally, in the specific case of veterinarians, efforts have also been made to promote mutual recognition, for instance the approval of a Program in Laboratory Animal Science and Medicine by VetCEE (Veterinary Continuous Education in Europe).
The New Guide and the European Framework: a Good Combination for a Successful Animal Care and Use Programme
Speaker: Javier Guillen, AAALAC International, Spain
Welfare Assessment and Severity Classification of Genetically Altered Rodents
Speaker: Anne Zintzsch, The Max Delbrück Center for Molecular Medicine, Berlin, Germany
Genetically Altered Mouse Models And New technologies
New technologies in model creation, humanised mouse models, and cryopreservation technologies
CRISPR/Cas9 in Rodent Animals: the Good, the Bad and the Ugly
Speaker: Guillaume Pavlović, Phenomin – Institut Clinique de la Souris, Illkirch, France
Strategies for Improving Reproducibility with In Vivo Studies
Speaker: Patrick Reilly, Phenomin – Institut Clinique de la Souris, Illkirch, France
Genetic Drift: What It Is and How to Minimize Its Impact on Your Research
Speaker: Catherine Hagan, The Jackson Laboratory, USA
Epigenetics: Beyond the Genetic Code
Speaker: Belén Pintado, Centro National Biotecnologia (CNB), Madrid, Spain
Cryopreservation of Mouse Sperm at -80 °C versus Liquid Nitrogen (LN2): A New and Simple Approach
Speaker: Marcello Raspa, National Research Council, Rome, Italy
Cryopreservation of germplasm enables researchers to dramatically reduce the use of animals and resources (3Rs approach) associated with unnecessary mouse colony maintenance, particularly of low use strains.
Sperm cryopreservation is a well-established technique for archiving mouse mutant strains and it is the most common approach in modern bio-repositories. We have investigated the possibility of cryopreserving mouse sperm in the absence of liquid nitrogen (LN2) using only a standard -80 °C freezer. Sperm fertility was also assessed after short-medium term storage at -80 °C. Sperm was frozen from two different genetically altered (GA) mouse lines on B6N background. The sperm was prepared for freezing in accordance with the MTG protocol which is widely used by the community. Sperm from two males was pooled and divided into 20 plastic (0.25 mL) semen straws. Ten straws were frozen using a standard freezing chamber filled with LN2. The other 10 straws were placed in a -80 °C freezer. After freezing the straws were maintained for up to 12 months at their freezing temperature i.e. -80 °C or LN2, as appropriate. Sperm was thawed at 7 days, 1 month, 3 months, 6 months, 9 months and 12 months after freezing and used in a standard IVF protocol in order access its fertility. Fertilisation rates, as judged by the generation of 2-cell embryo were not statistically different from the respective controls in LN2.
Our results show that sperm frozen using a standard -80 °C freezer can be maintained for at least 12 months without significant loss of fertility compared to sperm maintained in LN2. Furthermore, sperm maintained at -80 °C can be stored for long period in LN2 successfully. Sperm freezing in the absence of LN2 offers considerable advantages to laboratories that don’t have easy access to LN2 but still have a need to cryopreserve sperm. What is more, our results demonstrate that sperm can be held at -80 °C for at least 12 months without loss of potency. This will allow laboratories that don’t have access to LN2 to receive sperm held in repositories around the world. We believe it is important for all small laboratories to be able to cryopreserve and store mouse lines, even if they need to transfer the samples to a more secure repository at a later date. This is particularly important now that genome editing procedures (e.g. CRISPR/Cas9) permits the rapid generation of novel mouse models. Our hope is that this technique will remove another barrier to systemic sperm freezing in transgenic laboratories.
Genome Engineering with Integrases: Overexpression Strategies in the Mouse
Speaker: Ben Davies, Wellcome Trust Centre for Human Genetics, University of Oxford, UK
Animal Health Diagnostic Updates
What’s new in health monitoring and diagnostics, animal facility environments and animal research;
special emphasis will be placed on zebrafish health monitoring
Zebrafish Health Monitoring
Speaker: J-P Mocho, Daniovet, UK
Identifying and Managing Critical Control Points to Reduce the Risk of Adventitious Rodent Infections
Speaker: Guy B. Mulder, Charles River Laboratories, Wilmington, USA
Interceptor: “Interviewing” Air Exhaust Particles by PCR to Understand the Microbiological Status of Laboratory Rodents
Speaker: Gianpaolo Milite, Tecniplast, Italy
Husbandry and Biosecurity for Gnotobiotic Animals at the Centre for Gnotobiology at the Institut Pasteur
Speaker: Speaker: Marion Bérard, Institut Pasteur, France
Influence of the microbiota on animal models and monitoring baseline
Microbiome: Introduction, Pathophysiological Effects and Relevance for Laboratory Animals
Speaker: André Bleich, Institute for Laboratory Animal Science, Hannover Medical School, Germany
Humans and animals harbor 10-100 times more bacteria in their intestine than host cells. It is well established that these endogenous bacterial communities influence the host´s physiology considerably. Glrowth, aging, and various diseases (e.g., inflammatory bowel diseases, diabetes mellitus, arthritis, and obesity) are influenced by these communities, which are called microbiome, microbiota, or, historically, bacterial flora. This applies well to animal models. Phenotypes of these models, as well as the outcome of therapeutic interventions, depend considerably on the microbiome, and its influence on these models has outreached the effect of pathogens. Therefore, animal veterinarians and scientists should know how the microbiome of laboratory animal species is composed, which effect it might have, how its impact is currently being analysed, and how this research might cause a paradigm shift in health monitoring.
The Impact of the Microbiota on Rodent Models and How to Handle It
Speaker: Axel Kornerup Hansen, Department of Veterinary Disease Biology, University of Copenhagen, Denmark
Disease expression in a range of animal models, such as models of inflammatory bowel disease (IBD), diabetes, obesity, psychiatric disease and dermatitis correlate with the composition of the microbiota. Examples of commensal bacteria with an essential impact on animal model parameters are segmented filamentous bacteria (SFB), Akkermansia muciniphila, Prevotella spp. and bifidobacteria, which all have essential impact on the development or protection against inflammatory diseases in animal models. Stress, dietary fluctuations and other forms of uncontrolled impact may change the microbiota of the animal, and through this induce a change in the animal model, but newer studies indicate that a stable microbiota can be maintained even in IVC-housed mice. The simplest approach for handling microbiota impact on animal models is to screen animals when used for microbiota-sensitive studies and incorporate this information in data evaluation, thereby turning this uncontrolled variation into controlled variation. In the long run, we may be able to produce animals with less variation by microbiota standardization, e.g., by inoculation of tailor made microbiotas or feeding certain prebiotic diets. Ultimately, we may want to humanize the microbiota of the mouse and thereby the phenotype of the mouse itself. To do so, we still need to solve basic problems with the lack of immune priming from human bacteria in mice.
Translational Science Experiences
Experimental models to support translational science and medicine
New Translational Concepts in Liver Preservation for Transplantation; From Bench to Bedside
Speaker: René Tolba, University of Aachen Medical Center RWTH, Germany
Abstract: Coming soon
Patient-Derived Xenografts for Precision Oncology
Speaker: Andrea Bertotti, Laboratory of Translational Cancer Medicine, University of Torino, Italy
Almost twenty years after completion of the Human Genome Project, genetic alterations with causative roles in cancer have been identified. Inhibitors that target “druggable” mutant gene products have been developed. Finally, a number of these drugs have entered the clinic and are now used in patients.
Some fundamental issues need to be addressed before the informative potential of genome‐wide molecular surveys is translated into clinical benefit. First, given the heavy mutational load accrued during neoplastic progression, driver alterations are difficult to distinguish from passenger events. Second, next‐generation approaches are likely to identify previously neglected small subsets of very rare variants, which requires population‐scale studies for statistically solid genotype/response correlations. Third, not always does the presence of an oncogenic driver predict response. Therefore, treatment paradigm may not be decided on the sole basis of the mutational status of a single gene with potential cancer‐causing function, but rather on the basis of the heterogeneous context in which that mutation is found (‘precision medicine’).
All these challenges also apply to metastatic colorectal cancer (mCRC). Less than 20% of mCRC patients respond to anti‐EGFR antibodies – the gold standard targeted therapy – which calls for the identification of novel predictive biomarkers and suggests that uncharacterised drivers other than EGFR can sustain the growth of different CRC subsets.
The categorisation of tumour subpopulations featuring specific driver lesions, the validation of such lesions as targets, and the need for discrimination between ‘actionable’ opportunities and those with weak clinical transferability require reliable preclinical models. The scope of our studies is to refine genetic, mechanistic and therapeutic annotation of mCRC subsets using predictive in vivo models – namely, large collections of molecularly annotated patient‐derived tumour xenografts (PDXs) – as a means to systematically increase the success rate of rationally based clinical trials that emanate from preclinical findings.
Pathogenesis and Therapy of Hepatitis B Virus (HBV) Infection: Lessons from Animal Models
Speaker: Luca Guidotti, San Raffaele Scientific Institute, Italy
Owing to patients, experimentally infected chimpanzees, transgenic mice and other surrogate animal models (woodchucks, ground squirrels or ducks infected by naturally occurring hepadnaviruses), our understanding of hepatitis B virus (HBV) immunopathogenesis has greatly improved in the last few decades. It is generally acknowledged that HBV replicates noncytopathically in the hepatocyte, and that most of the clinical syndromes associated with this infection reflect the adaptive immune response, especially the virus-specific effector CD8+ T cell response. It is also evident that CD8+ T cell effector functions promote the resolution of HBV infection and that viral persistence results from the failure to induce or maintain these CD8+ T cell-dependent events. The means by which CD8+ T cells traffic, recognize hepatocellular antigens and deploy effector functions – and how such processes are affected by the anatomical conditions that characterize the normal or diseased liver – have been recently elucidated. This lecture discusses these scientific achievements that are related to HBV immunopathogenesis.
Highly Translational Phenotyping of Mice Bearing Disease-Relevant Mutations
Speaker: Maksym V. Kopanitsa, Charles River Discovery Research Services, Kuopio, Finland
Refinements in Experimental Procedures
Experimental design, research bias, and mathematical modelling
Refinements and Humane Endpoints in the Animal Model of Sepsis
Speaker: Manasi Nandi, Institute of Pharmaceutical Science, King’s College London, UK
Using Mathematics to Extract More from Physiological “Big Data”
Speaker: Manasi Nandi, Institute of Pharmaceutical Science, King’s College London, UK
More than Refinement – Improving the Validity and Reproducibility of Animal Research
Speaker: Hanno Würbel, Veterinary Public Health Institute, University of Bern, Switzerland
Introduction to Animal PBPK - Modelling and Prediction of PK-PD
Speaker: Janak Wedagedara, BSc, PhD, Senior Research Scientist, Certara
Back To Basics–Mouse and Rat
Biology of the mouse, identifying risk in research postmortem examination and tissue collection for diagnostics
Biology and Care of Laboratory Mice
Speaker: Jennifer C. Smith, Henry Ford Health System, Detroit, USA
Necropsy and Postmortem
Speaker: Christina Parkinson, Charles River Laboratories, Wilmington, USA