In Vivo Selection of a Computationally Designed SCHEMA AAV Library Yields a Novel Variant for Infection of Adult Neural Stem Cells in the SVZ

David S Ojala; Sabrina Sun; Jorge L Santiago-Ortiz; Mikhail G Shapiro; Philip A Romero; David V Schaffer

doi:10.1016/j.ymthe.2017.09.006

In Vivo Selection of a Computationally Designed SCHEMA AAV Library Yields a Novel Variant for Infection of Adult Neural Stem Cells in the SVZ

Mol Ther. 2018 Jan 3;26(1):304-319. doi: 10.1016/j.ymthe.2017.09.006. Epub 2017 Sep 8.

Authors

David S Ojala¹, Sabrina Sun¹, Jorge L Santiago-Ortiz¹, Mikhail G Shapiro², Philip A Romero³, David V Schaffer⁴

Affiliations

¹ Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA, USA.
² Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
³ Department of Biochemistry, University of Wisconsin, Madison, WI, USA.
⁴ Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA, USA; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA; The Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, CA, USA; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA. Electronic address: schaffer@berkeley.edu.

Abstract

Directed evolution continues to expand the capabilities of complex biomolecules for a range of applications, such as adeno-associated virus vectors for gene therapy; however, advances in library design and selection strategies are key to develop variants that overcome barriers to clinical translation. To address this need, we applied structure-guided SCHEMA recombination of the multimeric adeno-associated virus (AAV) capsid to generate a highly diversified chimeric library with minimal structural disruption. A stringent in vivo Cre-dependent selection strategy was implemented to identify variants that transduce adult neural stem cells (NSCs) in the subventricular zone. A novel variant, SCH9, infected 60% of NSCs and mediated 24-fold higher GFP expression and a 12-fold greater transduction volume than AAV9. SCH9 utilizes both galactose and heparan sulfate as cell surface receptors and exhibits increased resistance to neutralizing antibodies. These results establish the SCHEMA library as a valuable tool for directed evolution and SCH9 as an effective gene delivery vector to investigate subventricular NSCs.

Keywords: CNS; adeno-associated virus; capsid engineering; directed evolution; gene therapy; neural stem cell; subventricular zone.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Capsid Proteins / chemistry
Capsid Proteins / genetics
Dependovirus / classification
Dependovirus / genetics*
Dependovirus / ultrastructure
Galactose / metabolism
Gene Library
Gene Transfer Techniques
Genetic Engineering*
Genetic Therapy / methods
Genetic Vectors / genetics*
Genome, Viral
Heparitin Sulfate / metabolism
Humans
Imaging, Three-Dimensional
Lateral Ventricles / cytology*
Mice
Models, Molecular
Mutation
Neural Stem Cells / metabolism*
Protein Conformation
Transduction, Genetic*

Substances

Capsid Proteins
Heparitin Sulfate
Galactose

Grants and funding

R01 EY022975/EY/NEI NIH HHS/United States