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Human hematopoietic stem/progenitor cells modified by zinc-finger nucleases targeted to CCR5 control HIV-1 in vivo

Nature Biotechnology volume 28, pages 839–847 (2010)Cite this article

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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/IL2rγnull 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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Figure 1: ZFN-mediated disruption of CCR5 in CD34+ HSPCs.
Figure 2: Protection of human CD4+ T cells in peripheral blood of HIV-infected mice previously engrafted with ZFN-modified CD34+ HSPCs.
Figure 3: Effects of HIV-1 infection on human cells in HSPC-engrafted NSG mice.
Figure 4: HIV-1 infection selects for disrupted CCR5 alleles.
Figure 5: ZFN activity produces heterogeneous mutations in CCR5.
Figure 6: Control of HIV-1 replication in mice receiving ZFN-treated CD34+ HSPCs.

Change history

  • 20 July 2010

    In the version of this article initially published online, the callout to Figure 6b was written incorrectly as Figure 6c. Also, in Figure 2b, the label on the y axis was missing a “/” between CD4+ and CD8+. The errors have been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

We would like to thank A. Cuddihy, S. Ge, R. Hollis and N. Smiley for expert technical assistance; C. Lutzko, V. Garcia, R. Akkina, B. Torbett and M. McCune for advice regarding humanized mice; and M. McCune for communicating unpublished data. This work was supported by funding from the California HIV/AIDS Research Project (P.M.C.), The Saban Research Institute (V.T.), and the National Heart, Lung, and Blood Institute P01 HL73104 (G.M.C., D.B.K. and P.M.C.).

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Authors and Affiliations

  1. Keck School of Medicine of the University of Southern California, Los Angeles, California, USA

    Nathalia Holt & Paula M Cannon

  2. Sangamo BioSciences, Inc., Richmond, California, USA

    Jianbin Wang, Kenneth Kim, Geoffrey Friedman, Philip D Gregory & Michael C Holmes

  3. Childrens Hospital Los Angeles, Los Angeles, California, USA

    Xingchao Wang & Vanessa Taupin

  4. David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, California, USA

    Gay M Crooks & Donald B Kohn

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  1. Nathalia Holt
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Contributions

N.H. performed most of the experiments; J.W., K.K., G.F. and X.W. developed assays and analyzed samples; V.T. contributed to discussions; N.H., G.M.C., D.B.K., P.D.G., M.C.H. and P.M.C. designed the experiments and analyzed data; N.H. and P.M.C. wrote the manuscript.

Corresponding author

Correspondence to Paula M Cannon.

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J.W., K.K., G.F., P.D.G. and M.C.H. are employees of Sangamo BioSciences.

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Holt, N., Wang, J., Kim, K. et al. Human hematopoietic stem/progenitor cells modified by zinc-finger nucleases targeted to CCR5 control HIV-1 in vivo. Nat Biotechnol 28, 839–847 (2010). https://doi.org/10.1038/nbt.1663

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