Human hematopoietic stem/progenitor cells modified by zinc-finger nucleases targeted to CCR5 control HIV-1 in vivo
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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.
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Similar to CD4+ T cells, gene-modified HSCs were greatly enriched after infection.
“Similar to CD4+ T cells engraftment described earlier, the gene-modified cells were greatly enriched after infection [43].”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [43]
Holt and co-workers used nucleofection to achieve ZFN-mediated CCR5 disruption in human CD34+ HSCs.
“Holt and co-workers used nucleofection to achieve ZFN-mediated CCR5 disruption in human CD34+ HSCs.”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [43]
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- Authors
- Holt, N., Wang, J., Kim, K., Friedman, G., Wang, X., Taupin, V., Crooks, G. M., Kohn, D. B., Gregory, P. D., Holmes, M. C., & Cannon, P. M.
- Affiliations
- University of Southern California (US)
- Sangamo BioSciences (United States) (US)
- Children's Hospital Los Angeles (US)
- University of California, Los Angeles (US)
- Corresponding author
- Cannon, Paula M
- ORCID
- Venue
- Nature Biotechnology
- Published
- 2010-07-02
- Volume
- 28
- Issue
- 8
- Pages
- 839-847
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- PMC
- PMC3080757
- OpenAlex
- W2099106733
- Cited by
- 677
- References
- 75
- Citations by year
- 2026: 8 · 2025: 10 · 2024: 10 · 2023: 18 · 2022: 15 · 2021: 29 · 2020: 34 · 2019: 28 · 2018: 37 · 2017: 45 · 2016: 74 · 2015: 83 · 2014: 78 · 2013: 70 · 2012: 72
- FWCI
- 22.85
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- National Heart, Lung, and Blood Institute (P01HL073104, P01 HL73104)
- Topics
- Life Sciences › Immunology and Microbiology › Virology › HIV Research and Treatment
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Keywords
- HIV-1, CCR5, zinc finger nucleases, hematopoietic stem and progenitor cells, gene knockout, CCR5-tropic HIV-1, HIV drug resistance, autologous stem cell transplantation, CD4+ T cell depletion
- MeSH terms
- Animals, Endonucleases, Genetic Engineering, Hematopoietic Stem Cells, Humans, Stem Cells, T-Lymphocytes, HIV-1, HIV Infections, Zinc Fingers, Mice, SCID, Mice, Inbred NOD, Gene Deletion, Hematopoietic Stem Cell Transplantation, Receptors, CCR5, Mice
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