PMID- 26235624 OWN - NLM STAT- MEDLINE DCOM- 20160527 LR - 20220408 IS - 2211-1247 (Electronic) VI - 12 IP - 6 DP - 2015 Aug 11 TI - In Silico Reconstruction of the Viral Evolutionary Lineage Yields a Potent Gene Therapy Vector. PG - 1056-68 LID - S2211-1247(15)00759-7 [pii] LID - 10.1016/j.celrep.2015.07.019 [doi] AB - Adeno-associated virus (AAV) vectors have emerged as a gene-delivery platform with demonstrated safety and efficacy in a handful of clinical trials for monogenic disorders. However, limitations of the current generation vectors often prevent broader application of AAV gene therapy. Efforts to engineer AAV vectors have been hampered by a limited understanding of the structure-function relationship of the complex multimeric icosahedral architecture of the particle. To develop additional reagents pertinent to further our insight into AAVs, we inferred evolutionary intermediates of the viral capsid using ancestral sequence reconstruction. In-silico-derived sequences were synthesized de novo and characterized for biological properties relevant to clinical applications. This effort led to the generation of nine functional putative ancestral AAVs and the identification of Anc80, the predicted ancestor of the widely studied AAV serotypes 1, 2, 8, and 9, as a highly potent in vivo gene therapy vector for targeting liver, muscle, and retina. CI - Copyright (c) 2015 The Authors. Published by Elsevier Inc. All rights reserved. FAU - Zinn, Eric AU - Zinn E AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. FAU - Pacouret, Simon AU - Pacouret S AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. FAU - Khaychuk, Vadim AU - Khaychuk V AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. FAU - Turunen, Heikki T AU - Turunen HT AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. FAU - Carvalho, Livia S AU - Carvalho LS AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. FAU - Andres-Mateos, Eva AU - Andres-Mateos E AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. FAU - Shah, Samiksha AU - Shah S AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. FAU - Shelke, Rajani AU - Shelke R AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. FAU - Maurer, Anna C AU - Maurer AC AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. FAU - Plovie, Eva AU - Plovie E AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. FAU - Xiao, Ru AU - Xiao R AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. FAU - Vandenberghe, Luk H AU - Vandenberghe LH AD - Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA. Electronic address: luk_vandenberghe@meei.harvard.edu. LA - eng SI - GENBANK/KT235804 SI - GENBANK/KT235805 SI - GENBANK/KT235806 SI - GENBANK/KT235807 SI - GENBANK/KT235808 SI - GENBANK/KT235809 SI - GENBANK/KT235810 SI - GENBANK/KT235811 SI - GENBANK/KT235812 GR - DP1 EY023177/EY/NEI NIH HHS/United States GR - DP1 OD008267/OD/NIH HHS/United States GR - 5DP1EY023177-03/DP/NCCDPHP CDC HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20150730 PL - United States TA - Cell Rep JT - Cell reports JID - 101573691 SB - IM MH - Dependovirus/*genetics MH - *Gene Transfer Techniques MH - Genetic Therapy MH - Genetic Vectors/*genetics MH - Molecular Sequence Data PMC - PMC4536165 MID - NIHMS709331 COIS- CONFLICT OF INTEREST STATEMENT LHV and EZ are inventors on a patent describing some of the methods and reagents described here. LHV is inventor on several patents on gene therapy technologies. LHV is co-founder, shareholder, member of the Scientific Advisory Board, and consultant of GenSight Biologics, an ophthalmology gene therapy company. EDAT- 2015/08/04 06:00 MHDA- 2016/05/28 06:00 PMCR- 2016/08/11 CRDT- 2015/08/04 06:00 PHST- 2015/05/12 00:00 [received] PHST- 2015/06/26 00:00 [revised] PHST- 2015/07/09 00:00 [accepted] PHST- 2015/08/04 06:00 [entrez] PHST- 2015/08/04 06:00 [pubmed] PHST- 2016/05/28 06:00 [medline] PHST- 2016/08/11 00:00 [pmc-release] AID - S2211-1247(15)00759-7 [pii] AID - 10.1016/j.celrep.2015.07.019 [doi] PST - ppublish SO - Cell Rep. 2015 Aug 11;12(6):1056-68. doi: 10.1016/j.celrep.2015.07.019. Epub 2015 Jul 30.