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

Akhilesh Kumar

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

Akhilesh Kumar

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

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.

Original language	English (US)
Pages (from-to)	953-963
Number of pages	11
Journal	Stem Cell Reports
Volume	17
Issue number	4
DOIs	https://doi.org/10.1016/j.stemcr.2022.03.003
State	Published - Apr 12 2022

Bibliographical note

Funding Information:
This work is supported by funds from the National Institutes of Health, United States (R24OD021322, R01HL132891 and P51OD011106). The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: SIVmac316 open SpX from Dr. Ronald C. Desrosiers.

Funding Information:
This work is supported by funds from the National Institutes of Health, United States ( R24OD021322 , R01HL132891 and P51OD011106 ). The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: SIV mac 316 open SpX from Dr. Ronald C. Desrosiers.

Publisher Copyright:
© 2022 The Author(s)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

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

PubMed: MeSH publication types

Journal Article
Research Support, N.I.H., Extramural

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10.1016/j.stemcr.2022.03.003

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

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.",

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author = "Akhilesh Kumar",

note = "Funding Information: This work is supported by funds from the National Institutes of Health, United States (R24OD021322, R01HL132891 and P51OD011106). The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: SIVmac316 open SpX from Dr. Ronald C. Desrosiers. Funding Information: This work is supported by funds from the National Institutes of Health, United States ( R24OD021322 , R01HL132891 and P51OD011106 ). The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: SIV mac 316 open SpX from Dr. Ronald C. Desrosiers. Publisher Copyright: {\textcopyright} 2022 The Author(s)",

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AU - Kumar, Akhilesh

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N2 - 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.

AB - 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.

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KW - CRISPR-Cas9 gene editing

KW - HIV

KW - induced pluripotent stem cells

KW - macrophages

KW - nonhuman primates

KW - SIV

KW - T cells

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SN - 2213-6711

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JO - Stem Cell Reports

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

Abstract

Bibliographical note

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Keywords

PubMed: MeSH publication types

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