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Blood. 2016 May 19;127(20):2416-26. doi: 10.1182/blood-2015-09-672337. Epub 2016 Mar 15.

Long-term multilineage engraftment of autologous genome-edited hematopoietic stem cells in nonhuman primates.

Author information

1
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA;
2
Sangamo BioSciences, Richmond, CA; and.
3
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA; Department of Medicine and Department of Pathology, University of Washington, Seattle, WA.

Abstract

Genome editing in hematopoietic stem and progenitor cells (HSPCs) is a promising novel technology for the treatment of many human diseases. Here, we evaluated whether the disruption of the C-C chemokine receptor 5 (CCR5) locus in pigtailed macaque HSPCs by zinc finger nucleases (ZFNs) was feasible. We show that macaque-specific CCR5 ZFNs efficiently induce CCR5 disruption at levels of up to 64% ex vivo, 40% in vivo early posttransplant, and 3% to 5% in long-term repopulating cells over 6 months following HSPC transplant. These genome-edited HSPCs support multilineage engraftment and generate progeny capable of trafficking to secondary tissues including the gut. Using deep sequencing technology, we show that these ZFNs are highly specific for the CCR5 locus in primary cells. Further, we have adapted our clonal tracking methodology to follow individual CCR5 mutant cells over time in vivo, reinforcing that CCR5 gene-edited HSPCs are capable of long-term engraftment. Together, these data demonstrate that genome-edited HSPCs engraft, and contribute to multilineage repopulation after autologous transplantation in a clinically relevant large animal model, an important step toward the development of stem cell-based genome-editing therapies for HIV and potentially other diseases as well.

PMID:
26980728
PMCID:
PMC4874223
DOI:
10.1182/blood-2015-09-672337
[Indexed for MEDLINE]
Free PMC Article

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