Ayoub Mebarki, University of Calgary

Ayoub Mebarki, Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta, Canada. Blanca E. Callejas, Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada. Cynthia L. Swan, Vaccine and Infectious Disease Organization–International Vaccine Centre (VIDO-InterVac), University of Saskatchewan, Saskatoon, Saskatchewan, Canada. Ryan S. Noyce, Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta, Canada. Simon A. Hirota, Department of Physiology and Pharmacology, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada. Alyson A. Kelvin, Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta, Canada.

Discovering & Evolving

MPXV (formerly monkeypox virus) is a re-emerging zoonotic orthopoxvirus that caused multi-country outbreaks driven by newly described subclades Ib and IIb. Despite significant human-to-human transmission in recent outbreaks, MPXV remains an animal-origin virus, and establishment in new animal reservoirs could cause spillover events in new geographic areas. Canada has diverse wildlife populations and expanding contact zones between cities and natural habitats; however, the susceptibility of Canadian domestic, wildlife, and livestock to MPXV infection is not clear. This project addresses the potential expansion into new reservoir species by developing organoid models from Canadian species, such as the North American beaver (Castor canadensis), alongside wildlife surveillance to evaluate susceptibility to MPXV infection. In collaboration with the Human Organoid Innovation Hub at the University of Calgary, primary cells obtained through ethical partnerships will be dissociated and embedded in extracellular matrix–based three-dimensional cultures using species-appropriate differentiation media to generate organoids from tissues relevant to MPXV pathogenesis, including skin, lung, intestine, and testes. Organoids will be inoculated with MPXV to assess tissue tropism, replication kinetics, cellular damage, and viral localization. Transcriptomic analyses will be used to characterize species-specific host responses to infection. Vaccinia virus will serve as an orthopoxvirus control to establish baseline infection phenotypes, while organoids derived from permissive (CAST/EiJ) and resistant (C57BL/6) mouse strains will serve as experimental controls. Organoid-based susceptibility data will inform targeted wildlife surveillance through opportunistic sampling of relevant tissues and swabs screened by PCR, refining reservoir assessments, and strengthening Canada's capacity to anticipate and mitigate future mpox events.