Natural variations in AAVHSC16 significantly reduce liver tropism and maintain broad distribution to periphery and CNS

• Published in: Molecular therapy. Methods & clinical development 2022-09, Vol.26, p.224-238
• Main Authors: Smith, Laura J., Schulman, Lindsay A., Smith, Samantha, Van Lieshout, Laura, Barnes, Carmen M., Behmoiras, Liana, Scarpitti, Meghan, Kivaa, Monicah, Duong, Khanh L., Benard, Ludo O., Ellsworth, Jeff L., Avila, Nancy, Faulkner, Deiby, Hayes, April, Lotterhand, Jason, Rivas, Jose Israel, Sengooba, Arnold V., Tzianabos, Alec, Seymour, Albert B., Francone, Omar L.
• Format: Article
• Language: English
• Subjects: AAV; adeno-associated virus; gene therapy; glycan binding; Original; structure-function relationship; tropism/biodistribution
• ISSN: 2329-0501; 2329-0501
• DOI: 10.1016/j.omtm.2022.06.013

Adeno-associated viruses derived from human hematopoietic stem cells (AAVHSCs) are naturally occurring AAVs. Fifteen AAVHSCs have demonstrated broad biodistribution while displaying differences in transduction. We examine the structure-function relationships of these natural amino acid variations on cellular binding. We demonstrate that AAVHSC16 is the only AAVHSC that does not preferentially bind to terminal galactose. AAVHSC16 contains two unique amino acids, 501I and 706C, compared with other AAVHSCs. Through mutagenesis, we determined that residue 501 contributes to the lack of galactose binding. Structural analysis revealed that residue 501 is in proximity to the galactose binding pocket, hence confirming its functional role in galactose binding. Biodistribution analysis of AAVHSC16 indicated significantly less liver tropism in mice and non-human primates compared with other clade F members, likely associated with overall binding differences observed in vitro. AAVHSC16 maintained robust tropism to other key tissues in the peripheral and central nervous systems after intravenous injection, including to the brain, heart, and gastrocnemius. Importantly, AAVHSC16 did not induce elevated liver enzyme levels in non-human primates after intravenous injection at high doses. The unique glycan binding and tropism of AAVHSC16 makes this naturally occurring capsid an attractive candidate for therapies requiring less liver tropism while maintaining broad biodistribution. [Display omitted] Naturally occurring variations in AAVHSC16 that affect glycan binding lead to a unique in vivo biodistribution of lowered liver transduction with maintenance of broad systemic tropism in other peripheral and CNS tissues. AAVHSC16 displays a safe profile in NHPs with no elevation in alanine transferases after high-dose intravenous injection.