CRISPR/Cas9-Mediated CCR5 Ablation in Human Hematopoietic Stem/Progenitor Cells Confers HIV-1 Resistance In Vivo

Transplantation of hematopoietic stem cells (HSCs) with a naturally occurring CCR5 mutation confers a loss of detectable HIV-1 in the patient, making ablation of the CCR5 gene in HSCs an ideal therapy for an HIV-1 cure. Although CCR5 disruption has been attempted in CD4(+) T cells and hematopoietic stem/progenitor cells (HSPCs), efficient gene editing with high specificity and long-term therapeutic potential remains a major challenge for clinical translation. Here, we established a CRISPR/Cas9 gene editing system in human CD34(+) HSPCs and achieved efficient CCR5 ablation evaluated in long-term reconstituted NOD/Prkdc(scid)/IL-2R gamma(null) mice. The CCR5 disruption efficiency in our system remained robust in secondary transplanted repopulating hematopoietic cells. More importantly, an HIV-1 resistance effect was observed as indicated by significant reduction of virus titration and enrichment of human CD4(+) T cells. Hence, we successfully established a CRISPR/Cas9 mediated CCR5 ablating system in long-term HSCs, which confers HIV-1 resistance in vivo. Our study provides evidence for translating CCR5 gene-edited HSC transplantation for an HIV cure to the clinic.Beijing Science and Technology Major Project [D171100000517004]; National Natural Science Foundation of China [31521004]; National Science and Technology Support Project [2014BA102B01]; National High Technology Research and Development Program of China [2013ZX10001003003]; Guangdong Innovative and Entrepreneurial Research Team Program [2014ZT05S216]; Science and Technology Planning Project of Guangdong Province [2014B020226001]; Science and Technology Program of Guangzhou [2016B030232001]; Ministry of Education of China (111 Project); BeiHao Stem Cell and Regenerative Medicine Translational Research InstituteSCI(E)ARTICLE81782-17892