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

Nicole K. Paulk, Katja Pekrun, Erhua Zhu, Sean Nygaard, Bin Li, Jianpeng Xu, Kirk Chu, Christian Leborgne, Allison P. Dane, Annelise Haft, Yue Zhang, Feijie Zhang, Chris Morton, Marcus B. Valentine, Andrew M. Davidoff, Amit C. Nathwani, Federico Mingozzi, Markus Grompe, Ian E. Alexander, Leszek LisowskiMark A. Kay

Research output: Contribution to journalArticle

39 Scopus citations

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.

Original languageEnglish (US)
JournalMolecular Therapy
DOIs
StateAccepted/In press - 2017

Keywords

  • AAV
  • Evolution
  • Hepatocyte
  • Human
  • Library
  • Liver
  • Neutralization
  • Screen
  • Transduction

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

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    Paulk, N. K., Pekrun, K., Zhu, E., Nygaard, S., Li, B., Xu, J., Chu, K., Leborgne, C., Dane, A. P., Haft, A., Zhang, Y., Zhang, F., Morton, C., Valentine, M. B., Davidoff, A. M., Nathwani, A. C., Mingozzi, F., Grompe, M., Alexander, I. E., ... Kay, M. A. (Accepted/In press). Bioengineered AAV Capsids with Combined High Human Liver Transduction In Vivo and Unique Humoral Seroreactivity. Molecular Therapy. https://doi.org/10.1016/j.ymthe.2017.09.021