A simultaneous knockout knockin genome editing strategy in HSPCs potently inhibits CCR5- and CXCR4-tropic HIV-1 infection

Amanda M Dudek; William N Feist; Elena J Sasu; Sofia E Luna; Kaya Ben-Efraim; Rasmus O Bak; Alma-Martina Cepika; Matthew H Porteus

doi:10.1016/j.stem.2024.03.002

A simultaneous knockout knockin genome editing strategy in HSPCs potently inhibits CCR5- and CXCR4-tropic HIV-1 infection

Amanda M Dudek, William N Feist, Elena J Sasu, Sofia E Luna, Kaya Ben-Efraim, Rasmus O Bak, Alma-Martina Cepika, Matthew H Porteus^*

^*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

5 Citations (Scopus)

Abstract

Allogeneic hematopoietic stem and progenitor cell transplant (HSCT) of CCR5 null (CCR5Δ32) cells can be curative for HIV-1-infected patients. However, because allogeneic HSCT poses significant risk, CCR5Δ32 matched bone marrow donors are rare, and CCR5Δ32 transplant does not confer resistance to the CXCR4-tropic virus, it is not a viable option for most patients. We describe a targeted Cas9/AAV6-based genome editing strategy for autologous HSCT resulting in both CCR5- and CXCR4-tropic HIV-1 resistance. Edited human hematopoietic stem and progenitor cells (HSPCs) maintain multi-lineage repopulation capacity in vivo, and edited primary human T cells potently inhibit infection by both CCR5-tropic and CXCR4-tropic HIV-1. Modification rates facilitated complete loss of CCR5-tropic replication and up to a 2,000-fold decrease in CXCR4-tropic replication without CXCR4 locus disruption. This multi-factor editing strategy in HSPCs could provide a broad approach for autologous HSCT as a functional cure for both CCR5-tropic and CXCR4-tropic HIV-1 infections.

Original language	English
Journal	Cell Stem Cell
Volume	31
Issue	4
Pages (from-to)	499-518.e6
ISSN	1934-5909
DOIs	https://doi.org/10.1016/j.stem.2024.03.002
Publication status	Published - Apr 2024

Keywords

CCR5 knockout
CRISPR-Cas9
HIV
HIV restriction
autologous HSCT
cell therapy
functional cure
gene editing
hematopoietic stem cell transplant
homology-directed repair
Receptors, CXCR4/genetics
Humans
Receptors, CCR5/genetics
Gene Editing/methods
HIV Infections/genetics
HIV-1/genetics
Hematopoietic Stem Cells

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10.1016/j.stem.2024.03.002

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title = "A simultaneous knockout knockin genome editing strategy in HSPCs potently inhibits CCR5- and CXCR4-tropic HIV-1 infection",

abstract = "Allogeneic hematopoietic stem and progenitor cell transplant (HSCT) of CCR5 null (CCR5Δ32) cells can be curative for HIV-1-infected patients. However, because allogeneic HSCT poses significant risk, CCR5Δ32 matched bone marrow donors are rare, and CCR5Δ32 transplant does not confer resistance to the CXCR4-tropic virus, it is not a viable option for most patients. We describe a targeted Cas9/AAV6-based genome editing strategy for autologous HSCT resulting in both CCR5- and CXCR4-tropic HIV-1 resistance. Edited human hematopoietic stem and progenitor cells (HSPCs) maintain multi-lineage repopulation capacity in vivo, and edited primary human T cells potently inhibit infection by both CCR5-tropic and CXCR4-tropic HIV-1. Modification rates facilitated complete loss of CCR5-tropic replication and up to a 2,000-fold decrease in CXCR4-tropic replication without CXCR4 locus disruption. This multi-factor editing strategy in HSPCs could provide a broad approach for autologous HSCT as a functional cure for both CCR5-tropic and CXCR4-tropic HIV-1 infections.",

keywords = "CCR5 knockout, CRISPR-Cas9, HIV, HIV restriction, autologous HSCT, cell therapy, functional cure, gene editing, hematopoietic stem cell transplant, homology-directed repair, Receptors, CXCR4/genetics, Humans, Receptors, CCR5/genetics, Gene Editing/methods, HIV Infections/genetics, HIV-1/genetics, Hematopoietic Stem Cells",

author = "Dudek, {Amanda M} and Feist, {William N} and Sasu, {Elena J} and Luna, {Sofia E} and Kaya Ben-Efraim and Bak, {Rasmus O} and Alma-Martina Cepika and Porteus, {Matthew H}",

year = "2024",

month = apr,

doi = "10.1016/j.stem.2024.03.002",

language = "English",

volume = "31",

pages = "499--518.e6",

journal = "Cell Stem Cell",

issn = "1934-5909",

publisher = "Cell Press",

number = "4",

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A simultaneous knockout knockin genome editing strategy in HSPCs potently inhibits CCR5- and CXCR4-tropic HIV-1 infection. / Dudek, Amanda M; Feist, William N; Sasu, Elena J et al.
In: Cell Stem Cell, Vol. 31, No. 4, 04.2024, p. 499-518.e6.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - A simultaneous knockout knockin genome editing strategy in HSPCs potently inhibits CCR5- and CXCR4-tropic HIV-1 infection

AU - Dudek, Amanda M

AU - Feist, William N

AU - Sasu, Elena J

AU - Luna, Sofia E

AU - Ben-Efraim, Kaya

AU - Bak, Rasmus O

AU - Cepika, Alma-Martina

AU - Porteus, Matthew H

PY - 2024/4

Y1 - 2024/4

N2 - Allogeneic hematopoietic stem and progenitor cell transplant (HSCT) of CCR5 null (CCR5Δ32) cells can be curative for HIV-1-infected patients. However, because allogeneic HSCT poses significant risk, CCR5Δ32 matched bone marrow donors are rare, and CCR5Δ32 transplant does not confer resistance to the CXCR4-tropic virus, it is not a viable option for most patients. We describe a targeted Cas9/AAV6-based genome editing strategy for autologous HSCT resulting in both CCR5- and CXCR4-tropic HIV-1 resistance. Edited human hematopoietic stem and progenitor cells (HSPCs) maintain multi-lineage repopulation capacity in vivo, and edited primary human T cells potently inhibit infection by both CCR5-tropic and CXCR4-tropic HIV-1. Modification rates facilitated complete loss of CCR5-tropic replication and up to a 2,000-fold decrease in CXCR4-tropic replication without CXCR4 locus disruption. This multi-factor editing strategy in HSPCs could provide a broad approach for autologous HSCT as a functional cure for both CCR5-tropic and CXCR4-tropic HIV-1 infections.

AB - Allogeneic hematopoietic stem and progenitor cell transplant (HSCT) of CCR5 null (CCR5Δ32) cells can be curative for HIV-1-infected patients. However, because allogeneic HSCT poses significant risk, CCR5Δ32 matched bone marrow donors are rare, and CCR5Δ32 transplant does not confer resistance to the CXCR4-tropic virus, it is not a viable option for most patients. We describe a targeted Cas9/AAV6-based genome editing strategy for autologous HSCT resulting in both CCR5- and CXCR4-tropic HIV-1 resistance. Edited human hematopoietic stem and progenitor cells (HSPCs) maintain multi-lineage repopulation capacity in vivo, and edited primary human T cells potently inhibit infection by both CCR5-tropic and CXCR4-tropic HIV-1. Modification rates facilitated complete loss of CCR5-tropic replication and up to a 2,000-fold decrease in CXCR4-tropic replication without CXCR4 locus disruption. This multi-factor editing strategy in HSPCs could provide a broad approach for autologous HSCT as a functional cure for both CCR5-tropic and CXCR4-tropic HIV-1 infections.

KW - CCR5 knockout

KW - CRISPR-Cas9

KW - HIV

KW - HIV restriction

KW - autologous HSCT

KW - cell therapy

KW - functional cure

KW - gene editing

KW - hematopoietic stem cell transplant

KW - homology-directed repair

KW - Receptors, CXCR4/genetics

KW - Humans

KW - Receptors, CCR5/genetics

KW - Gene Editing/methods

KW - HIV Infections/genetics

KW - HIV-1/genetics

KW - Hematopoietic Stem Cells

UR - http://www.scopus.com/inward/record.url?scp=85189510061&partnerID=8YFLogxK

U2 - 10.1016/j.stem.2024.03.002

DO - 10.1016/j.stem.2024.03.002

M3 - Journal article

C2 - 38579682

SN - 1934-5909

VL - 31

SP - 499-518.e6

JO - Cell Stem Cell

JF - Cell Stem Cell

IS - 4

ER -

A simultaneous knockout knockin genome editing strategy in HSPCs potently inhibits CCR5- and CXCR4-tropic HIV-1 infection

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Keywords

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