title: Human hematopoietic stem/progenitor cells modified by zinc-finger nucleases targeted to CCR5 control HIV-1 in vivo.
abstract: CCR5 is the major HIV-1 co-receptor, and individuals homozygous for a 32-bp deletion in CCR5 are resistant to infection by CCR5-tropic HIV-1. Using engineered zinc-finger nucleases (ZFNs), we disrupted CCR5 in human CD34(+) hematopoietic stem/progenitor cells (HSPCs) at a mean frequency of 17% of the total alleles in a population. This procedure produces both mono- and bi-allelically disrupted cells. ZFN-treated HSPCs retained the ability to engraft NOD/SCID/IL2rgamma(null) mice and gave rise to polyclonal multi-lineage progeny in which CCR5 was permanently disrupted. Control mice receiving untreated HSPCs and challenged with CCR5-tropic HIV-1 showed profound CD4(+) T-cell loss. In contrast, mice transplanted with ZFN-modified HSPCs underwent rapid selection for CCR5(-/-) cells, had significantly lower HIV-1 levels and preserved human cells throughout their tissues. The demonstration that a minority of CCR5(-/-) HSPCs can populate an infected animal with HIV-1-resistant, CCR5(-/-) progeny supports the use of ZFN-modified autologous hematopoietic stem cells as a clinical approach to treating HIV-1.
authors.affiliation: Keck School of Medicine of the University of Southern California, Los Angeles, California, USA.
authors.forename: Nathalia
authors.initials: N
authors.lastname: Holt
authors.forename: Jianbin
authors.initials: J
authors.lastname: Wang
authors.forename: Kenneth
authors.initials: K
authors.lastname: Kim
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Friedman
authors.forename: Xingchao
authors.initials: X
authors.lastname: Wang
authors.forename: Vanessa
authors.initials: V
authors.lastname: Taupin
authors.forename: Gay M
authors.initials: GM
authors.lastname: Crooks
authors.forename: Donald B
authors.initials: DB
authors.lastname: Kohn
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.forename: Michael C
authors.initials: MC
authors.lastname: Holmes
authors.forename: Paula M
authors.initials: PM
authors.lastname: Cannon
mesh_terms: D000818:Animals; D004720:Endonucleases / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D005818:Genetic Engineering / Q000379:methods*; D015658:HIV Infections / Q000276:immunology / Q000628:therapy* / Q000821:virology; D015497:HIV-1 / Q000276:immunology / Q000378:metabolism; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D019713:Receptors, CCR5 / Q000235:genetics* / Q000378:metabolism; D013234:Stem Cells / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D016335:Zinc Fingers / Q000235:genetics* / Q000502:physiology
chemical_list: D019713:Receptors, CCR5; D004720:Endonucleases