title: Zinc-finger nuclease editing of human cxcr4 promotes HIV-1 CD4(+) T cell resistance and enrichment.
abstract: HIV-1-infected individuals can harbor viral isolates that can use CCR5, as well as CXCR4, for viral entry. To genetically engineer HIV-1 resistance in CD4(+) T cells, we assessed whether transient, adenovirus delivered zinc-finger nuclease (ZFN) disruption of genomic cxcr4 or stable lentiviral expression of short hairpin RNAs (shRNAs) targeting CXCR4 mRNAs provides durable resistance to HIV-1 challenge. ZFN-modification of cxcr4 in CD4(+) T cells was found to be superior to cell integrated lentivirus-expressing CXCR4 targeting shRNAs when CD4(+) T cells were challenged with HIV-1s that utilizes CXCR4 for entry. Cxcr4 disruption in CD4(+) T cells was found to be stable, conferred resistance, and provided for continued cell enrichment during HIV-1 infection in tissue culture and, in vivo, in peripheral blood mononuclear cell transplanted NSG mice. Moreover, HIV-1-infected mice with engrafted cxcr4 ZFN-modified CD4(+) T cells demonstrated lower viral levels in contrast to mice engrafted with unmodified CD4(+) T cells. These findings provide evidence that ZFN-mediated disruption of cxcr4 provides a selective advantage to CD4(+) T cells during HIV-1 infection.
authors.affiliation: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Jinyun
authors.initials: J
authors.lastname: Yuan
authors.forename: Jianbin
authors.initials: J
authors.lastname: Wang
authors.forename: Karen
authors.initials: K
authors.lastname: Crain
authors.forename: Colleen
authors.initials: C
authors.lastname: Fearns
authors.forename: Kenneth A
authors.initials: KA
authors.lastname: Kim
authors.forename: Kevin L
authors.initials: KL
authors.lastname: Hua
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.forename: Michael C
authors.initials: MC
authors.lastname: Holmes
authors.forename: Bruce E
authors.initials: BE
authors.lastname: Torbett
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D002478:Cells, Cultured; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D051379:Mice; D034741:RNA, Small Interfering / Q000235:genetics*; D019718:Receptors, CXCR4 / Q000235:genetics*; D016335:Zinc Fingers / Q000235:genetics
chemical_list: D034741:RNA, Small Interfering; D019718:Receptors, CXCR4; D004720:Endonucleases