Bioengineered AAV Capsids with Combined High Human Liver Transduction In Vivo and Unique Humoral Seroreactivity.

Autor:	Paulk NK; Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305, USA., Pekrun K; Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305, USA., Zhu E; Translational Vectorology Group, Children's Medical Research Institute, University of Sydney, Sydney, NSW, Australia., Nygaard S; Oregon Stem Cell Center, Oregon Health & Science University, Portland, OR 97239, USA., Li B; Oregon Stem Cell Center, Oregon Health & Science University, Portland, OR 97239, USA., Xu J; Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305, USA., Chu K; Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305, USA., Leborgne C; Genethon and INSERM U951, Evry, France., Dane AP; Department of Haematology, UCL Cancer Institute, London, UK., Haft A; Oregon Stem Cell Center, Oregon Health & Science University, Portland, OR 97239, USA., Zhang Y; Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305, USA., Zhang F; Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305, USA., Morton C; Department of Surgery, St. Jude Children's Research Hospital, Memphis, TN 38105, USA., Valentine MB; Cytogenetic Shared Resource, St. Jude Children's Research Hospital, Memphis, TN 38105, USA., Davidoff AM; Department of Surgery, St. Jude Children's Research Hospital, Memphis, TN 38105, USA., Nathwani AC; Department of Haematology, UCL Cancer Institute, London, UK; Department of Haematology and Katharine Dormandy Haemophilia Centre & Thrombosis Unit, Royal Free London NHS Foundation Trust Hospital, London, UK; National Health Services Blood and Transplant, Watford, UK., Mingozzi F; Genethon and INSERM U951, Evry, France; University Pierre and Marie Curie, Paris, France., Grompe M; Oregon Stem Cell Center, Oregon Health & Science University, Portland, OR 97239, USA., Alexander IE; Translational Vectorology Group, Children's Medical Research Institute, University of Sydney, Sydney, NSW, Australia., Lisowski L; Translational Vectorology Group, Children's Medical Research Institute, University of Sydney, Sydney, NSW, Australia; Military Institute of Hygiene and Epidemiology (MIHE), Puławy, Poland., Kay MA; Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305, USA. Electronic address: markay@stanford.edu.
Jazyk:	angličtina
Zdroj:	Molecular therapy : the journal of the American Society of Gene Therapy [Mol Ther] 2018 Jan 03; Vol. 26 (1), pp. 289-303. Date of Electronic Publication: 2017 Sep 25.
DOI:	10.1016/j.ymthe.2017.09.021
Abstrakt:	Existing recombinant adeno-associated virus (rAAV) serotypes for delivering in vivo gene therapy treatments for human liver diseases have not yielded combined high-level human hepatocyte transduction and favorable humoral neutralization properties in diverse patient groups. Yet, these combined properties are important for therapeutic efficacy. To bioengineer capsids that exhibit both unique seroreactivity profiles and functionally transduce human hepatocytes at therapeutically relevant levels, we performed multiplexed sequential directed evolution screens using diverse capsid libraries in both primary human hepatocytes in vivo and with pooled human sera from thousands of patients. AAV libraries were subjected to five rounds of in vivo selection in xenografted mice with human livers to isolate an enriched human-hepatotropic library that was then used as input for a sequential on-bead screen against pooled human immunoglobulins. Evolved variants were vectorized and validated against existing hepatotropic serotypes. Two of the evolved AAV serotypes, NP40 and NP59, exhibited dramatically improved functional human hepatocyte transduction in vivo in xenografted mice with human livers, along with favorable human seroreactivity profiles, compared with existing serotypes. These novel capsids represent enhanced vector delivery systems for future human liver gene therapy applications. (Copyright © 2017. Published by Elsevier Inc.)
Databáze:	MEDLINE
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