{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T15:11:30Z","timestamp":1788361890570,"version":"build-2803163510"},"reference-count":37,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2023,5,11]],"date-time":"2023-05-11T00:00:00Z","timestamp":1683763200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["R01EY027767"],"award-info":[{"award-number":["R01EY027767"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Vascular endothelial cells (VEC) are essential for retinal homeostasis and their dysfunction underlies pathogenesis in diabetic retinopathy (DR) and exudative age-related macular degeneration (AMD). Studies have shown that recombinant adeno-associated virus (rAAV) vectors are effective at delivering new genetic material to neural and glial cells within the retina, but targeting VECs remains challenging. To overcome this limitation, herein we developed rAAV capsid mutant vectors with improved tropism towards retinal VEC. rAAV2\/2, 2\/2[QuadYF-TV], and rAAV2\/9 serotype vectors (n = 9, capsid mutants per serotype) expressing GFP were generated by inserting heptameric peptides (7AA) designed to increase endothelial targeting at positions 588 (2\/2 and 2\/2[QuadYF-TV] or 589 (2\/9) of the virus protein (VP 1\u20133). The packaging and transduction efficiency of the vectors were assessed in HEK293T and bovine VECs using Fluorescence microscopy and flow cytometry, leading to the identification of one mutant, termed EC5, that showed improved endothelial tropism when inserted into all three capsid serotypes. Intra-ocular and intravenous administration of EC5 mutants in C57Bl\/6j mice demonstrated moderately improved transduction of the retinal vasculature, particularly surrounding the optic nerve head, and evidence of sinusoidal endothelial cell transduction in the liver. Most notably, intravenous administration of the rAAV2\/2[QuadYF-TV] EC5 mutant led to a dramatic and unexpected increase in cardiac muscle transduction.<\/jats:p>","DOI":"10.1371\/journal.pone.0285370","type":"journal-article","created":{"date-parts":[[2023,5,11]],"date-time":"2023-05-11T13:40:34Z","timestamp":1683812434000},"page":"e0285370","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":4,"title":["Improving retinal vascular endothelial cell tropism through rational rAAV capsid design"],"prefix":"10.1371","volume":"18","author":[{"given":"Ramesh","family":"Periasamy","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dwani D.","family":"Patel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sanford L.","family":"Boye","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shannon E.","family":"Boye","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7828-614X","authenticated-orcid":true,"given":"Daniel M.","family":"Lipinski","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2023,5,11]]},"reference":[{"issue":"262","key":"pone.0285370.ref001","doi-asserted-by":"crossref","first-page":"S1","DOI":"10.1038\/eye.2012.32","article-title":"Management of retinal vascular diseases: a patient-centric approach","volume":"2","author":"C.S. 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Homme","year":"2018","journal-title":"Front Physiol"},{"issue":"4","key":"pone.0285370.ref007","doi-asserted-by":"crossref","first-page":"606","DOI":"10.1038\/s41588-019-0351-9","article-title":"Retinal transcriptome and eQTL analyses identify genes associated with age-related macular degeneration","volume":"51","author":"R. Ratnapriya","year":"2019","journal-title":"Nat Genet"},{"issue":"4","key":"pone.0285370.ref008","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1128\/CMR.00008-08","article-title":"Gene therapy using adeno-associated virus vectors","volume":"21","author":"S. Daya","year":"2008","journal-title":"Clin Microbiol Rev"},{"issue":"9","key":"pone.0285370.ref009","doi-asserted-by":"crossref","first-page":"999","DOI":"10.1089\/hum.2009.086","article-title":"Human RPE65 gene therapy for Leber congenital amaurosis: persistence of early visual improvements and safety at 1 year","volume":"20","author":"A.V. 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