title: Terminal N-linked galactose is the primary receptor for adeno-associated virus 9.
abstract: Sialylated glycans serve as cell surface attachment factors for a broad range of pathogens. We report an atypical example, where desialylation increases cell surface binding and infectivity of adeno-associated virus (AAV) serotype 9, a human parvovirus isolate. Enzymatic removal of sialic acid, but not heparan sulfate or chondroitin sulfate, increased AAV9 transduction regardless of cell type. Viral binding and transduction assays on mutant Chinese hamster ovary (CHO) cell lines defective in various stages of glycan chain synthesis revealed a potential role for core glycan residues under sialic acid in AAV9 transduction. Treatment with chemical inhibitors of glycosylation and competitive inhibition studies with different lectins suggest that N-linked glycans with terminal galactosyl residues facilitate cell surface binding and transduction by AAV9. In corollary, resialylation of galactosylated glycans on the sialic acid-deficient CHO Lec2 cell line with different sialyltransferases partially blocked AAV9 transduction. Quantitative analysis of AAV9 binding to parental, sialidase-treated or sialic acid-deficient mutant CHO cells revealed a 3-15-fold increase in relative binding potential of AAV9 particles upon desialylation. Finally, pretreatment of well differentiated human airway epithelial cultures and intranasal instillation of recombinant sialidase in murine airways enhanced transduction efficiency of AAV9 by >1 order of magnitude. Taken together, the studies described herein provide a molecular basis for low infectivity of AAV9 in vitro and a biochemical strategy to enhance gene transfer by AAV9 vectors in general.
authors.affiliation: Gene Therapy Center, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
authors.forename: Shen
authors.initials: S
authors.lastname: Shen
authors.forename: Kelli D
authors.initials: KD
authors.lastname: Bryant
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Brown
authors.forename: Scott H
authors.initials: SH
authors.lastname: Randell
authors.forename: Aravind
authors.initials: A
authors.lastname: Asokan
mesh_terms: D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D000229:Dependovirus / Q000235:genetics / Q000378:metabolism*; D005690:Galactose / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D019158:N-Acetylneuraminic Acid / Q000235:genetics / Q000378:metabolism*; D011991:Receptors, Virus / Q000378:metabolism*; D012799:Sialyltransferases / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods; D053585:Virus Attachment
chemical_list: D011991:Receptors, Virus; D012799:Sialyltransferases; D019158:N-Acetylneuraminic Acid; D005690:Galactose