Delivering genes across the blood-brain barrier: LY6A, a novel cellular receptor for AAV-PHP.B capsids

Autoři: Qin Huang aff001;  Ken Y. Chan aff001;  Isabelle G. Tobey aff001;  Yujia Alina Chan aff001;  Tim Poterba aff001;  Christine L. Boutros aff002;  Alejandro B. Balazs aff002;  Richard Daneman aff003;  Jonathan M. Bloom aff001;  Cotton Seed aff001;  Benjamin E. Deverman aff001
Působiště autorů: Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA, United States of America aff001;  Ragon Institute of MGH, MIT, and Harvard, Cambridge, MA, United States of America aff002;  Departments of Neurosciences and Pharmacology, University of California, San Diego, La Jolla, CA, United States of America aff003;  Department of Pharmacology, University of California, San Diego, La Jolla, CA, United States of America aff004
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0225206


The engineered AAV-PHP.B family of adeno-associated virus efficiently delivers genes throughout the mouse central nervous system. To guide their application across disease models, and to inspire the development of translational gene therapy vectors for targeting neurological diseases in humans, we sought to elucidate the host factors responsible for the CNS tropism of the AAV-PHP.B vectors. Leveraging CNS tropism differences across 13 mouse strains, we systematically determined a set of genetic variants that segregate with the permissivity phenotype, and rapidly identified LY6A as an essential receptor for the AAV-PHP.B vectors. Interfering with LY6A by CRISPR/Cas9-mediated Ly6a disruption or with blocking antibodies reduced transduction of mouse brain endothelial cells by AAV-PHP.eB, while ectopic expression of Ly6a increased AAV-PHP.eB transduction of HEK293T and CHO cells by 30-fold or more. Importantly, we demonstrate that this newly discovered mode of AAV binding and transduction can occur independently of other known AAV receptors. These findings illuminate the previously reported species- and strain-specific tropism characteristics of the AAV-PHP.B vectors and inform ongoing efforts to develop next-generation AAV vehicles for human CNS gene therapy.

Klíčová slova:

Capsids – Cell binding – Cell binding assay – Central nervous system – Endothelial cells – Galactose – Immunohistochemistry techniques – Mammalian genomics


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2019 Číslo 11