Endothelial retargeting of AAV9 in vivo

Abstract Background Adeno-associated virus (AAV) is favorable choice for gene transfer and editing applications in vivo, due to its low integration rate immunogenicity, as well a variety of natural engineered serotypes providing different tissue cell type tropism. Endothelial cells, however, are remarkably resistant transduction by AAVs. Purpose The aim this study retarget wild AAV9 vectors using combination polyamidoamine dendrimers (PAMAM) peptides with endothelial affinity. Methods An endothelial-affine peptide sequence was obtained via bio-panning phage display library on culture cells. Second-generation PAMAM were linked the PEG linker. coated these modified (G2CNN) immediately before vivo applications. Results mTmG-mice or mTmG-pigs utilized reporter organisms, where red-to green fluorescence change upon Cre expression displays cellular resolution transduction. encoding AAV G2CNN application systemically (mice) locally (pigs) transduced over 30% CD31+ cells skeletal muscle heart. Functional relevance retargeting assessed systemic injections three transgenes: 1) artificial adhesion molecule (S1FG). In mice, S1FG increased leukocyte cremaster vasculature (day 10 post injection). 2) Anti-inflammatory Annexin A1 (Anxa1). ApoE−/− mice high-fat diet, Anxa1 reduced long term recruitment carotid artery 28 3) sgRNA targeting vasodilatory enzyme eNOS (endothelial nitric oxide synthase). conditional Cas9 transgenic delivered caused increase blood pressure 56 Conclusion retargeted efficiently heart vivo. Accordingly, vascular atherosclerosis models, may enable transfer. Funding Acknowledgement Type funding sources: Public grant(s) – National budget only. Main source(s): Bundesministerium für Bildung und ForschungDeutsche Forschungsgemeinschaft