A dirofilariasis mouse model for heartworm preclinical research

Marriott, A. E.; Dagley, J. L.; Hegde, S.; Steven, A.; Fricks, C.; DiCosty, U.; Mansour, A.; Campbell, E. J.; Wilson, C. M.; Gusovsky, F.; Ward, S. A.; Hong, W. D.; O'Neill, P.; Moorhead, A.; McCall, S.; McCall, J. W.; Taylor, M. J.; Turner, J. D.

doi:10.1101/2023.04.03.535321

bioRxiv · 10.1101/2023.04.03.535321

A dirofilariasis mouse model for heartworm preclinical research

Marriott, A. E.·Dagley, J. L.·Hegde, S.·Steven, A.·Fricks, C.·DiCosty, U.·Mansour, A.·Campbell, E. J.·Wilson, C. M.·Gusovsky, F.·Ward, S. A.·Hong, W. D.·O'Neill, P.·Moorhead, A.·McCall, S.·McCall, J. W.·Taylor, M. J.·Turner, J. D.

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Abstract

Use of experimental cats and dogs in veterinary heartworm preclinical drug research is increasing. As a potential alternative primary in vivo heartworm preventative drug screen, we assessed lymphopenic mice with ablation of the interleukin-2/7 common gamma chain ({gamma}c) as susceptible to the larval development phase of D. immitis. Non-obese diabetic (NOD) Severe Combined ImmunoDeficient (SCID){gamma}c-/-(NSG / NXG) mice consistently yielded viable D. immitis larvae at 2-4 weeks post-infection across multiple experiments, different batches of infectious larvae inoculates, different isolates of D. immitis and at independent laboratories. Mice did not display any overt clinical signs associated with infection up to 4 weeks. Developing larvae were found in subcutaneous and muscle fascia tissues, the natural site of this stage of heartworm in dogs. Larvae retrieved from NSG / NXG mice were mid-L4 stage of development. Compared with 14-day in vitro propagated larvae, in vivo derived L4 were significantly larger and contained expanded Wolbachia endobacteria titres, determined by QPCR and Fluorescent in situ Hybridisation (FISH). We established an ex vivo 6-day L4 paralytic screening system against nematodicidal agents (moxidectin, levamisole) which highlighted discrepancies in relative drug sensitivities in comparison with in vitro reared L4 D. immitis. We demonstrated effective depletion of Wolbachia by 70-90% in D. immitis L4 following 2-7 day oral in vivo exposures of NSG / NXG infected mice with doxycycline or the rapid-acting investigational anti-Wolbachia drug, AWZ1066S. We validated the NSG / NXG mouse model as a filaricide drug screen by in vivo treatments with single injections of moxidectin, which mediated 60-88% reduction in L4 larvae at 14-28 days. Future adoption of the mouse model as a first-line efficacy screen will benefit end-user laboratories conducting research and development of novel heartworm preventatives via increased access, rapid turnaround and reduced costs whilst simultaneously decreasing need for experimental cat or dog use.

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Marriott, A. E., Dagley, J. L., Hegde, S., Steven, A., Fricks, C., DiCosty, U., Mansour, A., Campbell, E. J., Wilson, C. M., Gusovsky, F., Ward, S. A., Hong, W. D., O'Neill, P., Moorhead, A., McCall, S., McCall, J. W., Taylor, M. J., Turner, J. D.. 2023-04-03. A dirofilariasis mouse model for heartworm preclinical research. https://doi.org/10.1101/2023.04.03.535321

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