Selection of an Efficient AAV Vector for Robust CNS Transgene Expression

Killian S Hanlon; Jonah C Meltzer; Tetyana Buzhdygan; Ming J Cheng; Miguel Sena-Esteves; Rachel E Bennett; Timothy P Sullivan; Roshanak Razmpour; Yi Gong; Carrie Ng; Josette Nammour; Daniela Maiz; Simon Dujardin; Servio H Ramirez; Eloise Hudry; Casey A Maguire

doi:10.1016/j.omtm.2019.10.007

Selection of an Efficient AAV Vector for Robust CNS Transgene Expression

Mol Ther Methods Clin Dev. 2019 Oct 23:15:320-332. doi: 10.1016/j.omtm.2019.10.007. eCollection 2019 Dec 13.

Authors

Killian S Hanlon^{1

2

3}, Jonah C Meltzer^{3

4}, Tetyana Buzhdygan^{5

6

7}, Ming J Cheng^{2

3}, Miguel Sena-Esteves⁸, Rachel E Bennett^{3

4}, Timothy P Sullivan^{5

6

7}, Roshanak Razmpour^{5

6

7}, Yi Gong^{2

3}, Carrie Ng^{2

3}, Josette Nammour^{2

3}, Daniela Maiz^{3

4}, Simon Dujardin^{3

4}, Servio H Ramirez^{5

6

7}, Eloise Hudry^{3

4}, Casey A Maguire^{2

3}

Affiliations

¹ Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA.
² Molecular Neurogenetics Unit, Department of Neurology, Massachusetts General Hospital, Charlestown, MA 02129.
³ Harvard Medical School, Boston, MA 02115, USA.
⁴ Alzheimer's Disease Research Laboratory, Department of Neurology, Massachusetts General Hospital, Charlestown, MA 02129, USA.
⁵ Department of Pathology and Laboratory Medicine, Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, USA.
⁶ Shriners Hospital's Pediatric Research Center, Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, USA.
⁷ Center for Substance Abuse Research, Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, USA.
⁸ University of Massachusetts Medical School, Worcester, MA 01655, USA.

Abstract

Adeno-associated virus (AAV) capsid libraries have generated improved transgene delivery vectors. We designed an AAV library construct, iTransduce, that combines a peptide library on the AAV9 capsid with a Cre cassette to enable sensitive detection of transgene expression. After only two selection rounds of the library delivered intravenously in transgenic mice carrying a Cre-inducible fluorescent protein, we flow sorted fluorescent cells from brain, and DNA sequencing revealed two dominant capsids. One of the capsids, termed AAV-F, mediated transgene expression in the brain cortex more than 65-fold (astrocytes) and 171-fold (neurons) higher than the parental AAV9. High transduction efficiency was sex-independent and sustained in two mouse strains (C57BL/6 and BALB/c), making it a highly useful capsid for CNS transduction of mice. Future work in large animal models will test the translation potential of AAV-F.

Keywords: AAV capsid library; AAV vector; adeno-associated virus vector; central nervous system; gene delivery; gene therapy; transduction efficiency.

Abstract

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