title: An essential receptor for adeno-associated virus infection.
abstract: Adeno-associated virus (AAV) vectors are currently the leading candidates for virus-based gene therapies because of their broad tissue tropism, non-pathogenic nature and low immunogenicity. They have been successfully used in clinical trials to treat hereditary diseases such as haemophilia B (ref. 2), and have been approved for treatment of lipoprotein lipase deficiency in Europe. Considerable efforts have been made to engineer AAV variants with novel and biomedically valuable cell tropisms to allow efficacious systemic administration, yet basic aspects of AAV cellular entry are still poorly understood. In particular, the protein receptor(s) required for AAV entry after cell attachment remains unknown. Here we use an unbiased genetic screen to identify proteins essential for AAV serotype 2 (AAV2) infection in a haploid human cell line. The most significantly enriched gene of the screen encodes a previously uncharacterized type I transmembrane protein, KIAA0319L (denoted hereafter as AAV receptor (AAVR)). We characterize AAVR as a protein capable of rapid endocytosis from the plasma membrane and trafficking to the trans-Golgi network. We show that AAVR directly binds to AAV2 particles, and that anti-AAVR antibodies efficiently block AAV2 infection. Moreover, genetic ablation of AAVR renders a wide range of mammalian cell types highly resistant to AAV2 infection. Notably, AAVR serves as a critical host factor for all tested AAV serotypes. The importance of AAVR for in vivo gene delivery is further highlighted by the robust resistance of Aavr(-/-) (also known as Au040320(-/-) and Kiaa0319l(-/-)) mice to AAV infection. Collectively, our data indicate that AAVR is a universal receptor involved in AAV infection.
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, 299 Campus Drive, Stanford, California 94305, USA.
authors.forename: S
authors.initials: S
authors.lastname: Pillay
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Medicine, Oregon Health &Science University, 3181 Sam Jackson Park Road, Portland, Oregon 97239-3098, USA.
authors.forename: N L
authors.initials: NL
authors.lastname: Meyer
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, 299 Campus Drive, Stanford, California 94305, USA.
authors.forename: A S
authors.initials: AS
authors.lastname: Puschnik
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Medicine, Oregon Health &Science University, 3181 Sam Jackson Park Road, Portland, Oregon 97239-3098, USA.
authors.forename: O
authors.initials: O
authors.lastname: Davulcu
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, 299 Campus Drive, Stanford, California 94305, USA.
authors.forename: J
authors.initials: J
authors.lastname: Diep
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Medicine, Oregon Health &Science University, 3181 Sam Jackson Park Road, Portland, Oregon 97239-3098, USA.|Shriners Hospital for Children, 3101 Sam Jackson Park Road, Portland, Oregon 97239, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Ishikawa
authors.affiliation: Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, Netherlands.
authors.forename: L T
authors.initials: LT
authors.lastname: Jae
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, 299 Campus Drive, Stanford, California 94305, USA.
authors.forename: J E
authors.initials: JE
authors.lastname: Wosen
authors.affiliation: Department of Comparative Medicine, Stanford University School of Medicine, 287 Campus Drive, Stanford, California 94305, USA.
authors.forename: C M
authors.initials: CM
authors.lastname: Nagamine
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Medicine, Oregon Health &Science University, 3181 Sam Jackson Park Road, Portland, Oregon 97239-3098, USA.
authors.forename: M S
authors.initials: MS
authors.lastname: Chapman
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, 299 Campus Drive, Stanford, California 94305, USA.
authors.forename: J E
authors.initials: JE
authors.lastname: Carette
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D002460:Cell Line; D000229:Dependovirus / Q000145:classification / Q000187:drug effects / Q000502:physiology*; D004705:Endocytosis / Q000187:drug effects; D005260:Female; D017353:Gene Deletion; D015316:Genetic Therapy / Q000379:methods; D058507:Host Specificity; D006801:Humans; D008297:Male; D051379:Mice; D010322:Parvoviridae Infections / Q000378:metabolism* / Q000821:virology*; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D011991:Receptors, Virus / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D056189:Viral Tropism / Q000187:drug effects; D053586:Virus Internalization / Q000187:drug effects; D021601:trans-Golgi Network / Q000187:drug effects
chemical_list: D000906:Antibodies; C541251:KIAA0319L protein, human; C000605136:KIAA0319L protein, mouse; D011956:Receptors, Cell Surface; D011991:Receptors, Virus
references.citation: Trends Biochem Sci. 2005 Jun;30(6):309-17
references.pmid: 15950875
references.citation: J Virol. 1998 Feb;72(2):1438-45
references.pmid: 9445046
references.citation: Hum Mol Genet. 2000 Jul 1;9(11):1641-9
references.pmid: 10861291
references.citation: J Cell Biol. 2004 Apr;165(1):111-22
references.pmid: 15078902
references.citation: Nature. 2015 Apr 9;520(7546):186-91
references.pmid: 25830891
references.citation: J Virol. 2005 Jan;79(1):609-14
references.pmid: 15596854
references.citation: PLoS One. 2009 Aug 06;4(8):e6529
references.pmid: 19657394
references.citation: Nature. 2011 Nov 30;481(7379):81-4
references.pmid: 22139420
references.citation: Mol Ther. 2008 Jun;16(6):1073-80
references.pmid: 18414476
references.citation: N Engl J Med. 2011 Dec 22;365(25):2357-65
references.pmid: 22149959
references.citation: Virol J. 2013 Mar 06;10:74
references.pmid: 23497173
references.citation: J Virol. 2004 Jul;78(13):7186-98
references.pmid: 15194795
references.citation: Nat Protoc. 2012 Jan 05;7(1):171-92
references.pmid: 22222791
references.citation: Curr Opin Cell Biol. 2011 Aug;23(4):404-12
references.pmid: 21450449
references.citation: Trends Cell Biol. 2011 Mar;21(3):159-67
references.pmid: 21183348
references.citation: Nat Rev Genet. 2014 Jul;15(7):445-51
references.pmid: 24840552
references.citation: J Virol. 2000 Jan;74(2):992-6
references.pmid: 10623762
references.citation: Nat Med. 1999 Jan;5(1):71-7
references.pmid: 9883842
references.citation: Nat Rev Mol Cell Biol. 2004 Feb;5(2):121-32
references.pmid: 15040445
references.citation: Nat Rev Mol Cell Biol. 2003 Mar;4(3):202-12
references.pmid: 12612639
references.citation: Curr Opin Lipidol. 2012 Aug;23(4):310-20
references.pmid: 22691709
references.citation: Gene Ther. 2012 Jun;19(6):649-58
references.pmid: 22357511
references.citation: Cell Host Microbe. 2011 Dec 15;10(6):563-76
references.pmid: 22177561
references.citation: Mol Psychiatry. 2011 Apr;16(4):365-82
references.pmid: 20956978
references.citation: Traffic. 2011 Aug;12(8):963-71
references.pmid: 21463457
references.citation: Nat Protoc. 2013 Nov;8(11):2281-308
references.pmid: 24157548
references.citation: Nature. 2011 Aug 24;477(7364):340-3
references.pmid: 21866103
references.citation: Nature. 2014 Feb 20;506(7488):382-6
references.pmid: 24390344
references.citation: Philos Trans R Soc Lond B Biol Sci. 2015 Feb 5;370(1661):20140035
references.pmid: 25533093
references.citation: Science. 2014 Jun 27;344(6191):1506-10
references.pmid: 24970085