LY6A was identified as an essential receptor for AAV-PHP.B vectors, with ectopic expression of Ly6a increasing AAV-PHP.eB transduction of human cells by 30-fold.
LY6A is an essential cellular receptor for AAV-PHP.B vectors, enabling efficient gene delivery across the blood-brain barrier.
p-value: p=<0.001
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.
Huang et al. (Thu,) conducted a other in Gene therapy delivery across the blood-brain barrier. AAV-PHP.eB and Ly6a expression vs. AAV9 or nonpermissive strains/untransfected cells was evaluated on AAV-PHP.eB binding and transduction (p=<0.001). LY6A was identified as an essential receptor for AAV-PHP.B vectors, with ectopic expression of Ly6a increasing AAV-PHP.eB transduction of human cells by 30-fold.