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

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 and Matthew H. Porteus
Cell stem cell, Vol.31(4), pp.499-518.e6
04/04/2024
DOI: 10.1016/j.stem.2024.03.002
PMCID: PMC11212398
PMID: 38579682

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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. [Display omitted] •HSPC editing to develop HSC transplantation to treat HIV•Edited HSPCs and T cells retain normal functionality•CCR5 knockout causes complete loss of CCR5-tropic HIV replication•Multi-factor knockin causes a multi-log-fold decrease in CXCR4-tropic HIV replication Dudek, Feist et al. describe a gene editing strategy for a functional cure for HIV-1 through hematopoietic stem and progenitor cell transplant. CCR5-locus knockin of multiple inhibitory protein factors to generate combinatorial resistance in terminally differentiated progeny allows strong inhibition of both CCR5-tropic and CXCR4-tropic HIV-1 strains.
autologous HSCT CCR5 knockout cell therapy CRISPR-Cas9 functional cure gene editing hematopoietic stem cell transplant HIV HIV restriction homology-directed repair

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