PMID- 28789965 OWN - NLM STAT- MEDLINE DCOM- 20180604 LR - 20240315 IS - 1873-4995 (Electronic) IS - 0168-3659 (Print) IS - 0168-3659 (Linking) VI - 262 DP - 2017 Sep 28 TI - Application of polyploid adeno-associated virus vectors for transduction enhancement and neutralizing antibody evasion. PG - 348-356 LID - S0168-3659(17)30772-1 [pii] LID - 10.1016/j.jconrel.2017.08.005 [doi] AB - Adeno-associated virus (AAV) vectors have been used successfully in clinical trials for patients with hemophilia or blindness, but pre-existing neutralizing antibodies (Nab) are common in the general population and exclude many patients from clinical trials. Exploration of effective strategies to enhance AAV transduction and escape from Nab activity is still imperative. Previous studies have shown the compatibility of capsids from AAV serotypes and homology of recognition sites of AAV Nab located on different capsid subunits from one virion. In this study, we co-transfected AAV2 and AAV8 helper plasmids at different ratios (3:1, 1:1 and 1:3) to assemble haploid capsids and study both their transduction efficiency and Nab escape activity. After muscular injection, all of the haploid viruses induced higher transduction than their parental AAV vectors (2- to 9-fold over AAV2), with the highest of these being the haploid vector AAV2/8 3:1. After systemic administration, a 4-fold higher transduction in the liver was observed with haploid AAV2/8 1:3 than that with AAV8 alone. We then packaged the therapeutic factor IX cassette into haploid AAV2/8 1:3 capsids and injected them into FIX knockout mice via the tail vein. Higher FIX expression and improved phenotypic correction were achieved with the haploid AAV2/8 1:3 virus vector when compared to that of AAV8. Additionally, the haploid virus AAV2/8 1:3 was able to escape AAV2 neutralization and did not increase capsid antigen presentation capacity when compared to AAV8. To improve the Nab evasion ability of the haploid virus, we produced the triploid vector AAV2/8/9 by co-transfecting AAV2, AAV8 and AAV9 helper plasmids at a ratio of 1:1:1. After systemic administration, a 2-fold higher transduction in the liver was observed with the triploid vector AAV2/8/9 than that with AAV8. Nab analysis demonstrated that the triploid AAV2/8/9 vector was able to escape Nab activity from mouse sera immunized with parental serotypes. These results indicate that polyploid viruses might potentially acquire advantages from parental serotypes for enhancement of AAV transduction and evasion of Nab recognition without increasing capsid antigen presentation in target cells. Polyploid AAV vectors can be generated from any AAV serotype, whether natural, rational, library derived or a combination thereof, providing a novel strategy that should be explored in future clinical trials in patients with neutralizing antibodies. CI - Copyright (c) 2017. Published by Elsevier B.V. FAU - Chai, Zheng AU - Chai Z AD - Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, United States. FAU - Sun, Junjiang AU - Sun J AD - Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, United States. FAU - Rigsbee, Kelly Michelle AU - Rigsbee KM AD - Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, United States. FAU - Wang, Mei AU - Wang M AD - Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, United States; Institute of Hematology, Chinese Academy of Medical Sciences, Tianjin, China. FAU - Samulski, R Jude AU - Samulski RJ AD - Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, United States; Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, United States. Electronic address: rjs@med.unc.edu. FAU - Li, Chengwen AU - Li C AD - Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, United States; Department of Pediatrics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, United States. Electronic address: chengwen_li@med.unc.edu. LA - eng GR - R01 DK084033/DK/NIDDK NIH HHS/United States GR - R01 AI117408/AI/NIAID NIH HHS/United States GR - R01 AI072176/AI/NIAID NIH HHS/United States GR - P30 CA016086/CA/NCI NIH HHS/United States GR - R01 HL125749/HL/NHLBI NIH HHS/United States GR - U54 CA151652/CA/NCI NIH HHS/United States GR - P01 HL112761/HL/NHLBI NIH HHS/United States PT - Journal Article DEP - 20170805 PL - Netherlands TA - J Control Release JT - Journal of controlled release : official journal of the Controlled Release Society JID - 8607908 RN - 0 (Antibodies, Neutralizing) SB - IM MH - Animals MH - Antibodies, Neutralizing/immunology MH - Capsid MH - Cell Line MH - Cell Line, Tumor MH - Cell Proliferation MH - Dependovirus/*genetics MH - Female MH - *Genetic Vectors MH - Humans MH - Liver/metabolism MH - Mice MH - Mice, Inbred C57BL MH - Mice, Knockout MH - Muscle, Skeletal/metabolism MH - T-Lymphocytes MH - Transduction, Genetic PMC - PMC5819605 MID - NIHMS940937 OTO - NOTNLM OT - AAV OT - Gene therapy OT - Neutralizing antibody OT - Polyploid OT - Transduction COIS- Disclosure of conflict of interest R. Jude Samulski is the founder, and a shareholder, at Asklepios BioPharmaceutical. He receives research support through the University of North Carolina from Asklepios BioPharmaceutical. He holds patents that have been licensed by UNC to Asklepios Biopharmaceutical, for which he receives royalties. He has consulted for Baxter Healthcare and has received payment for speaking. EDAT- 2017/08/10 06:00 MHDA- 2018/06/05 06:00 PMCR- 2018/09/28 CRDT- 2017/08/10 06:00 PHST- 2017/04/22 00:00 [received] PHST- 2017/07/18 00:00 [revised] PHST- 2017/08/02 00:00 [accepted] PHST- 2017/08/10 06:00 [pubmed] PHST- 2018/06/05 06:00 [medline] PHST- 2017/08/10 06:00 [entrez] PHST- 2018/09/28 00:00 [pmc-release] AID - S0168-3659(17)30772-1 [pii] AID - 10.1016/j.jconrel.2017.08.005 [doi] PST - ppublish SO - J Control Release. 2017 Sep 28;262:348-356. doi: 10.1016/j.jconrel.2017.08.005. Epub 2017 Aug 5.