Generation of SIV-resistant T cells and macrophages from nonhuman primate induced pluripotent stem cells with edited CCR5 locus

Saritha S D'Souza; Akhilesh Kumar; Jason Weinfurter; Mi Ae Park; John Maufort; Lihong Tao; HyunJun Kang; Samuel T Dettle; Thaddeus Golos; James A Thomson; Matthew R Reynolds; Igor Slukvin

doi:10.1016/j.stemcr.2022.03.003

Generation of SIV-resistant T cells and macrophages from nonhuman primate induced pluripotent stem cells with edited CCR5 locus

Stem Cell Reports. 2022 Apr 12;17(4):953-963. doi: 10.1016/j.stemcr.2022.03.003. Epub 2022 Mar 31.

Authors

Affiliations

¹ Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI 53715, USA.
² Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI 53706, USA.
³ Morgridge Institute for Research, Madison, USA.
⁴ Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Comparative Biosciences, University of Wisconsin-Madison, Madison, WI 53706 USA; Department of Obstetrics and Gynecology, University of Wisconsin-Madison, Madison, WI 53705 USA.
⁵ Morgridge Institute for Research, Madison, USA; Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA 93106, USA.
⁶ Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI 53706, USA.
⁷ Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Cell and Regenerative Biology, University of Wisconsin-Madison, Madison, WI 53706, USA; Department of Pathology and Laboratory Medicine, University of Wisconsin-Madison, Madison, WI 53792, USA. Electronic address: [email protected].

Abstract

Adoptive therapies with genetically modified somatic T cells rendered HIV resistance have shown promise for AIDS therapy. A renewable source of HIV-resistant human T cells from induced pluripotent stem cells (iPSCs) would further facilitate and broaden the applicability of these therapies. Here, we report successful targeting of the CCR5 locus in iPSCs generated from T cells (T-iPSCs) or fibroblasts (fib-iPSCs) from Mauritian cynomolgus macaques (MCM), using CRISPR-Cas9 technology. We found that CCR5 editing does not affect hematopoietic and T cell differentiation potentials of fib-iPSCs. However, T-iPSCs with edited CCR5 lost their capacity to differentiate into CD4⁺CD8⁺ T cells while maintaining myeloid differentiation potential. T cells and macrophages produced from CCR5-edited MCM iPSCs did not support replication of the CCR5-tropic simian immunodeficiency viruses SIVmac239 (T cell tropic) and SIVmac316 (macrophage-tropic). Overall, these studies provide a platform for further exploration of AIDS therapies based on gene-edited iPSCs in a nonhuman primate model.

Keywords: CCR5; CRISPR-Cas9 gene editing; HIV; SIV; T cells; induced pluripotent stem cells; macrophages; nonhuman primates.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Acquired Immunodeficiency Syndrome*
Animals
CD8-Positive T-Lymphocytes
Induced Pluripotent Stem Cells*
Macaca fascicularis
Macrophages
Receptors, CCR5 / genetics

Substances

Receptors, CCR5

Abstract

Publication types

MeSH terms

Substances

Grants and funding