Nine new awards in our fifth year of 3Rs knowledge transfer

The funding scheme is a critical part of the NC3Rs’ commitment to tackling the lag in uptake of 3Rs methods, by enabling exchange of models, tools and technologies between research groups.

The Skills and Knowledge Transfer grants are an annual competition with awards of up to £75k available over a maximum of 24 months. Applications made to the scheme include both a ‘developer’ and ‘end-user(s)’ of the model, tool or technology; either party can be the primary applicant. Of the nine awards made in the 2021 competition, five were made to developers and four to end-users. This year the Skills and Knowledge Transfer scheme evolved to enable SMEs to apply as lead applicant, facilitating greater connectivity between academics and industry. Three of the new awards include partnerships between SMEs, CROs and academia.

Our collaboration with Cancer Research UK continues to support researchers transferring 3Rs approaches in the cancer research field. One co-funded award was made this year to replace the use of xenograft mice in some breast cancer metastasis studies.

This year’s awards are summarised below:

Dr Hannah Harrison, University of Manchester, £75,381 (co-funded with CRUK) – Replacing in vivo models with the Quasi vivo system to investigate metastatic site priming by tumour cells.

Hannah will take up and adapt an in vitro method from her collaborators at the University of Pisa to investigate signalling events that prime secondary tumour formation during metastasis. Hannah will co-culture breast cancer cells with other cell types from major sites of breast cancer metastasis, including bone, liver and lung cells, which are cultured in separate chambers with media flowing constantly through the whole system. This method will replace some in vivo experiments in Hannah’s research where mice are injected with cancer cells to study metastasis by determining where secondary tumours develop in the animal.

Dr Matthew Parker, University of Portsmouth, £63,587 – A non-protected larval zebrafish model for the investigation of novel strategies to protect against nerve agent-induced toxicity and seizures.

As a postdoctoral researcher on an NC3Rs-funded Project grant, Matthew developed non-invasive imaging procedures to visualise seizures in larval zebrafish using lightsheet microscopy. Based on current thinking, at this early stage the larvae are not considered capable of suffering and they therefore provide a partial replacement for the use of other animals.

Matthew will adapt the imaging method for use with standard confocal microscopes, enabling wider uptake by other groups. He will then transfer this method to Dstl to identify novel treatments for nerve agent poisoning. Matthew will work with Dstl to create high-throughput assays that accurately quantify convulsive behaviour, replacing procedures in mice associated with high levels of suffering. Dstl will also promote the method more widely to military and defence organisations in Australia, America and Canada.

Dr Ben Newland, Cardiff University, £74,526 – Establishing the novel ex-vivo focal demyelination model of Multiple Sclerosis in other laboratories to reduce and replace live animal use.

In vivo studies of multiple sclerosis are associated with severe suffering, including weakness, forelimb and hindlimb paralysis and incontinence, requiring specialist husbandry and care. Ben and colleagues have developed a method using demyelinating agents to induce focal MS-like lesions in ex vivo tissue slice cultures from embryonic mice. Up to six brain slices and ten spinal cord slices can be isolated from each embryo and the method ensures both healthy and diseased tissue is present in the slice culture, similarly to how MS presents in patients. Ben will transfer the method to two research groups at Queen’s University Belfast and create detailed protocols and training videos to enable others to take up the method.

Professor Simon Schultz, Imperial College London, £75,552 – Cerebral organoid models for optical investigation of neural circuit dynamics in neurodegenerative diseases.

Brain recordings are taken in vivo, typically in mice, to better understand how accumulation of tau and amyloid beta affects patients in Alzheimer’s disease. These studies require a window to be surgically implanted in the skull to be able to visualise the brain. Simon aims to replace some of this research in his laboratory by combining 3Rs methods from research groups based in the MRC Laboratory of Molecular Biology and University College London. Simon will use established protocols to generate cerebral organoids from human iPSCs and characterise neural circuits using multiphoton calcium imaging to determine how these are affected by altered tau pathology. This will demonstrate further utility of the cerebral organoids and their potential to replace neuroimaging in mice.

Dr Patrick Varga-Weisz, University of Essex, £64,941 – Acquisition of the Drosophila model system to understand mechanisms of innate immunity regulation by chromatin dynamics.

Patrick has previously identified a critical component in the innate immunity response of the gut using in vivo infection studies in mice. To explore this further, without the need for mouse models, he will collaborate with Dr Dominique Ferrandon at the Institute de Biologie Moléculare et Cellulaire to set up the infrastructure needed to work with Drosophila at the University of Essex. Dominique will train Patrick and colleagues in key techniques to study innate immunity in Drosophila, which is highly conserved in vertebrates and invertebrates. Patrick will then work with collaborators in Brazil, where Drosophila is not widely used as a model, to further promote non-protected multicellular models in basic scientific research.

Dr Anne Herrmann, University of Liverpool, £66,435 – Go North: Expanding the use of the chick embryo model in cancer research to a multiuser facility at the University of Strathclyde.

Through an NC3Rs David Sainsbury Fellowship, Anne developed a platform using chick embryos to study cancer initiation, growth and metastasis. She has generated standard operating procedures for multiple cancer types, including breast, head and neck, lung, colon, and pancreatic cancers replacing some xenograft mouse studies in these areas. To enable further replacement beyond the University of Liverpool, Anne will transfer protocols and cell lines to the University of Strathclyde and train a specialised technician to set up the infrastructure needed for chick embryo experiments. Researchers at Strathclyde will then be invited to trial using chick embryos in their own studies reducing the barriers to uptake.

Dr Laura Randle, Liverpool John Moores University, £74,093 – Determining the efficacy and safety of cancer chemotherapeutics for cholangiocarcinoma (CCA) using Human Precision Cut Tissue Slices (hPCTS).

Bile duct cancer is typically studied in mice, either by implanting tumour tissue from patients or introducing relevant genetic mutations through breeding programmes. The percentage of animals that possess the mutation as a result of these programmes is small, increasing the number of animals needing to be bred for studies. Precision cut tissue slices can be cultured ex vivo using human tissue, retaining all cell types in a 3D environment. Laura will establish this method in her laboratory receiving training, expertise and advice from her European and American collaborators on working with human tissue. Laura will confirm the clinical relevance of the method and validate the model by analysing tissue response to chemotherapy drugs currently used in patient treatment.

Dr Virginia Marugan-Hernandez, Royal Veterinary College, £73,136 – Transfer of an in vitro model for the evaluation of anticoccidial compounds.

Virginia will transfer an in vitro screening method to test new interventions for coccidiosis, a parasitic disease that infects the intestines of chickens, to SALUVET-innova, an SME spun-out of the Complutense University of Madrid. This will enable SALUVET-innova to screen out ineffective compounds so that they do not progress into in vivo studies. The in vitro platform can also inform treatment strategies, including dosing timings, further reducing the need for in vivo testing in chickens. The in vitro assay requires parasites to be propagated so to enable wider 3Rs impact, SALUVET-innova will offer the assay as a pre-screening service to other companies and research groups who may not have the infrastructure necessary to produce the parasites.

Dr Joseph Turner, Liverpool School of Tropical Medicine, £75,686 – Adoption of a mouse model of veterinary filariasis for preclinical drug testing.

Veterinary filariasis, or heartworm disease, is a parasitic disease that predominantly affects cats and dogs. New interventions for heartworm are currently tested in cats and dogs, which can cause adverse events due to the infection, from reduced activity and laboured breathing to cardiopulmonary embolism. With NC3Rs funding, Joe has developed a refined model of heartworm that mimics the early stages of infection using immunodeficient mice. The parasites are propagated under the skin reducing the likelihood of adverse events as experiments in mice last for a few weeks rather than up to 12 months in cats and dogs. Joe will transfer the refined mouse model to two companies, who between them published over 50% of heartworm drug research publications in the last three years, reducing the number of heartworm drug testing procedures needing to be performed in cats and dogs.