@article{eef5ca07cf5048db92fe4f8eb26d4b2d,
title = "Bioengineered AAV Capsids with Combined High Human Liver Transduction In Vivo and Unique Humoral Seroreactivity",
abstract = "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. Paulk et al. performed sequential directed evolution screens of AAV capsid libraries in primary human hepatocytes in vivo and against human sera from thousands of patients. Resultant capsid variants AAV-NP40, AAV-NP59, and AAV-NP84 exhibit unique seroreactivity and significantly increase functional human hepatocyte transduction in vivo compared to existing hepatotropic serotypes.",
keywords = "AAV, evolution, hepatocyte, human, library, liver, neutralization, screen, transduction",
author = "Paulk, {Nicole K.} and Katja Pekrun and Erhua Zhu and Sean Nygaard and Bin Li and Jianpeng Xu and Kirk Chu and Christian Leborgne and Dane, {Allison P.} and Annelise Haft and Yue Zhang and Feijie Zhang and Chris Morton and Valentine, {Marcus B.} and Davidoff, {Andrew M.} and Nathwani, {Amit C.} and Federico Mingozzi and Markus Grompe and Alexander, {Ian E.} and Leszek Lisowski and Kay, {Mark A.}",
note = "Funding Information: The authors wish to acknowledge Derek Pouchnik and Mark Wildung of the WSU Laboratory for Biotechnology & Analysis for sequencing help. N.K.P. was supported by postdoctoral fellowships from the National Heart Lung & Blood Institute (grant F32-HL119059), the American Liver Foundation Hans Popper Memorial Fellowship, and the Stanford Dean's Fellowship. This work was supported by grants to M.A.K. from the NIH (grant R01-HL092096); to M.V. from the NIH (grant P30-CA21765); to A.D. from the NIH (grant R01-HL073838), the ALSAC St. Jude Children's Research Hospital, and the Assisi Foundation of Memphis; to M.G. from the NIH (grant R01-DK048252); to F.M. from the ERC (grant CoG-617432); and to I.E.A. from the Australian NHMRC (grant APP-1008021). This project was supported by a NIH Shared Instrumentation Grant (S10-OD01058001-A1) from the NCRR with significant contribution from Stanford's Beckman Center, as well as the OHSU Flow Cytometry Shared Resource. The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of the various funding bodies or universities involved. Packaging plasmids for any of the new capsids described herein must be obtained through a material transfer agreement (MTA) with Stanford University. Funding Information: The authors wish to acknowledge Derek Pouchnik and Mark Wildung of the WSU Laboratory for Biotechnology & Analysis for sequencing help. N.K.P. was supported by postdoctoral fellowships from the National Heart Lung & Blood Institute (grant F32-HL119059 ), the American Liver Foundation Hans Popper Memorial Fellowship , and the Stanford Dean{\textquoteright}s Fellowship . This work was supported by grants to M.A.K. from the NIH (grant R01-HL092096 ); to M.V. from the NIH (grant P30-CA21765 ); to A.D. from the NIH (grant R01-HL073838 ), the ALSAC St. Jude Children{\textquoteright}s Research Hospital , and the Assisi Foundation of Memphis ; to M.G. from the NIH (grant R01-DK048252 ); to F.M. from the ERC (grant CoG-617432 ); and to I.E.A. from the Australian NHMRC (grant APP-1008021 ). This project was supported by a NIH Shared Instrumentation Grant ( S10-OD01058001-A1 ) from the NCRR with significant contribution from Stanford{\textquoteright}s Beckman Center , as well as the OHSU Flow Cytometry Shared Resource . The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of the various funding bodies or universities involved. Packaging plasmids for any of the new capsids described herein must be obtained through a material transfer agreement (MTA) with Stanford University. Publisher Copyright: {\textcopyright} 2017",
year = "2018",
month = jan,
day = "3",
doi = "10.1016/j.ymthe.2017.09.021",
language = "English (US)",
volume = "26",
pages = "289--303",
journal = "Molecular Therapy",
issn = "1525-0016",
publisher = "Nature Publishing Group",
number = "1",
}