Xu Lei, Yang Huan, Gao Yang, Chen Zeyu, Xie Liangfu, Liu Yulin, Liu Ying, Wang Xiaobao, Li Hanwei, Lai Weifeng, He Yuan, Yao Anzhi, Ma Liying, Shao Yiming, Zhang Bin, Wang Chengyan, Chen Hu, Deng Hongkui
Department of Hematopoietic Stem Cell Transplantation, 307 Hospital of PLA, Beijing 100071, China; Cell and Gene Therapy Center, Academy of Military Medical Sciences, Beijing 100850, China.
Department of Cell Biology, School of Basic Medical Sciences, Peking University Stem Cell Research Center; State Key Laboratory of Natural and Biomimetic Drugs, Peking University Health Science Center, Peking University, Beijing 100191, China; MOE Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100191, China; Shenzhen Stem Cell Engineering Laboratory, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen 518055, China.
Mol Ther. 2017 Aug 2;25(8):1782-1789. doi: 10.1016/j.ymthe.2017.04.027. Epub 2017 May 17.
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/IL-2Rγ 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.
移植具有天然存在的CCR5突变的造血干细胞（HSC）可使患者体内检测不到HIV-1，因此在HSC中敲除CCR5基因是治愈HIV-1的理想疗法。尽管已尝试在CD4 T细胞和造血干/祖细胞（HSPC）中破坏CCR5，但具有高特异性和长期治疗潜力的高效基因编辑仍然是临床转化的主要挑战。在此，我们在人CD34 HSPC中建立了CRISPR/Cas9基因编辑系统，并在长期重建的NOD/Prkdc/IL-2Rγ小鼠中评估了高效的CCR5敲除。我们系统中的CCR5破坏效率在二次移植的再填充造血细胞中仍然很强劲。更重要的是，观察到了HIV-1抗性效应，表现为病毒滴定显著降低和人CD4 T细胞富集。因此，我们成功地在长期HSC中建立了CRISPR/Cas9介导的CCR5敲除系统，该系统在体内赋予HIV-1抗性。我们的研究为将CCR5基因编辑的HSC移植用于治愈HIV的疗法转化至临床提供了证据。
