A Comprehensive Atlas of AAV Tropism in the Mouse

Christopher J Walkey; Kathy J Snow; Jote Bulcha; Aaron R Cox; Alexa E Martinez; M Cecilia Ljungberg; Denise G Lanza; Marco De Giorgi; Marcel A Chuecos; Michele Alves-Bezerra; Carlos Flores Suarez; Sean M Hartig; Susan G Hilsenbeck; Chih-Wei Hsu; Ethan Saville; Yaned Gaitan; Jeff Duryea Jr; Seth Hannigan; Mary E Dickinson; Oleg Mirochnitchenko; Dan Wang; Cathleen M Lutz; Jason D Heaney; Guangping Gao; Stephen A Murray; William R Lagor

doi:10.1016/j.ymthe.2025.01.041

A Comprehensive Atlas of AAV Tropism in the Mouse

Mol Ther. 2025 Jan 25:S1525-0016(25)00043-7. doi: 10.1016/j.ymthe.2025.01.041. Online ahead of print.

Authors

Christopher J Walkey¹, Kathy J Snow², Jote Bulcha³, Aaron R Cox⁴, Alexa E Martinez¹, M Cecilia Ljungberg⁵, Denise G Lanza⁶, Marco De Giorgi¹, Marcel A Chuecos¹, Michele Alves-Bezerra¹, Carlos Flores Suarez¹, Sean M Hartig⁷, Susan G Hilsenbeck⁸, Chih-Wei Hsu¹, Ethan Saville², Yaned Gaitan², Jeff Duryea Jr², Seth Hannigan², Mary E Dickinson¹, Oleg Mirochnitchenko⁹, Dan Wang¹⁰, Cathleen M Lutz², Jason D Heaney⁶, Guangping Gao³, Stephen A Murray², William R Lagor¹¹

Affiliations

¹ Department of Integrative Physiology, Baylor College of Medicine, Houston, TX 77030, USA.
² The Jackson Laboratory, 600 Main Street, Bar Harbor, ME 04609 USA.
³ Horae Gene Therapy Center and Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01605, USA; Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, MA 01605, USA.
⁴ Section of Diabetes, Endocrinology, and Metabolism, Department of Medicine, Baylor College of Medicine, Houston, TX 77030, USA.
⁵ Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030, USA; Duncan Neurological Research Institute, Texas Children's Hospital, Houston, TX 77030, USA.
⁶ Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas, USA.
⁷ Section of Diabetes, Endocrinology, and Metabolism, Department of Medicine, Baylor College of Medicine, Houston, TX 77030, USA; Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
⁸ Dan L. Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, Texas, USA.
⁹ Office of Research Infrastructure Programs, Division of Program Coordination, Planning, and Strategic Initiatives, Office of the Director, National Institutes of Health, Bethesda, MD USA.
¹⁰ Horae Gene Therapy Center and Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01605, USA; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USA.
¹¹ Department of Integrative Physiology, Baylor College of Medicine, Houston, TX 77030, USA. Electronic address: [email protected].

PMID: 39863928
DOI: 10.1016/j.ymthe.2025.01.041

Abstract

Gene therapy with Adeno-Associated Virus (AAV) vectors requires knowledge of their tropism within the body. Here we analyze the tropism of ten naturally occurring AAV serotypes (AAV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrh10 and AAVrh74) following systemic delivery into male and female mice. A transgene expressing ZsGreen and Cre recombinase was used to identify transduction in a cell-dependent manner based on fluorescence. Cre-driven activation of tdTomato fluorescence offered superior sensitivity for transduced cells. All serotypes except AAV3B and AAV4 had high liver tropism. Fluorescence activation revealed transduction of unexpected tissues, including adrenals, testes and ovaries. Rare transduced cells within tissues were also readily visualized. Biodistribution of AAV genomes correlated with fluorescence, except in immune tissues. AAV4 was found to have a pan-endothelial tropism while also targeting pancreatic beta cells. This public resource enables selection of the best AAV serotypes for basic science and preclinical applications in mice.