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Analysis of recombinant adeno-associated viral vector shedding in sheep following intracoronary delivery

Gene Therapy volume 26, pages 399–406 (2019)Cite this article

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Abstract

Differences between mouse and human hearts pose a significant limitation to the value of small animal models when predicting vector behavior following recombinant adeno-associated viral (rAAV) vector-mediated cardiac gene therapy. Hence, sheep have been adopted as a preclinical animal, as they better model the anatomy and cardiac physiological processes of humans. There is, however, no comprehensive data on the shedding profile of rAAV in sheep following intracoronary delivery, so as to understand biosafety risks in future preclinical and clinical applications. In this study, sheep received intracoronary delivery of rAAV serotypes 2/6 (2 × 1012 vg), 2/8, and 2/9 (1 × 1013 vg) at doses previously administered in preclinical and clinical trials. This was followed by assessment over 96 h to examine vector shedding in urine, feces, nasal mucus, and saliva samples. Vector genomes were detected via real-time quantitative PCR in urine and feces up to 48 and 72 h post vector delivery, respectively. Of these results, functional vector particles were only detected via a highly sensitive infectious replication assay in feces samples up to 48 h following vector delivery. We conclude that rAAV-mediated gene transfer into sheep hearts results in low-grade shedding of non-functional vector particles for all excreta samples, except in the case of feces, where functional vector particles are present up to 48 h following vector delivery. These results may be used to inform containment and decontamination guidelines for large animal dealings, and to understand the biosafety risks associated with future preclinical and clinical uses of rAAV.

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Acknowledgements

Our work was supported by a National Health and Medical Research Council of Australia Project Grant (APP1128864) to EK, Future Leader Fellowship (ID 100463) from the National Heart Foundation of Australia (JJHC) and a Sydney Medical School Foundation Fellowship (JJHC). MF was supported by the MyWestmead Early Career Research Scholarship and Arab Bank Postgraduate Research Scholarship from the Westmead Medical Research Foundation. We acknowledge the facilities and services provided by the Westmead Institute for Medical Research and Westmead Research Hub Core facilities including the following: Microscopy, Flow Cytometry, and Genomics. We also acknowledge the facilities and staff of the Western Sydney Local Health District Large Animal Facility.

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Authors and Affiliations

  1. Sydney Medical School, The University of Sydney, Sydney, NSW, Australia

    Melad Farraha, James Chong & Eddy Kizana

  2. Center for Heart Research, The Westmead Institute for Medical Research, Sydney, NSW, Australia

    Melad Farraha, Thi Y. L. Le, Sindhu Igoor, Renuka Rao, Cindy Kok, James Chong & Eddy Kizana

  3. Department of Cardiology, Westmead Hospital, Sydney, NSW, Australia

    Michael A. Barry, Juntang Lu, Jim Pouliopoulos, James Chong & Eddy Kizana

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Farraha, M., Barry, M.A., Lu, J. et al. Analysis of recombinant adeno-associated viral vector shedding in sheep following intracoronary delivery. Gene Ther 26, 399–406 (2019). https://doi.org/10.1038/s41434-019-0097-0

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