De Ravin Suk See, Brault Julie, Meis Ronald J, Liu Siyuan, Li Linhong, Pavel-Dinu Mara, Lazzarotto Cicera R, Liu Taylor, Koontz Sherry M, Choi Uimook, Sweeney Colin L, Theobald Narda, Lee GaHyun, Clark Aaron B, Burkett Sandra S, Kleinstiver Benjamin P, Porteus Matthew H, Tsai Shengdar, Kuhns Douglas B, Dahl Gary A, Headey Stephen, Wu Xiaolin, Malech Harry L
Genetic Immunotherapy Section, Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD.
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Blood. 2021 May 13;137(19):2598-2608. doi: 10.1182/blood.2020008503.
Lentivector gene therapy for X-linked chronic granulomatous disease (X-CGD) has proven to be a viable approach, but random vector integration and subnormal protein production from exogenous promoters in transduced cells remain concerning for long-term safety and efficacy. A previous genome editing-based approach using Streptococcus pyogenes Cas9 mRNA and an oligodeoxynucleotide donor to repair genetic mutations showed the capability to restore physiological protein expression but lacked sufficient efficiency in quiescent CD34+ hematopoietic cells for clinical translation. Here, we report that transient inhibition of p53-binding protein 1 (53BP1) significantly increased (2.3-fold) long-term homology-directed repair to achieve highly efficient (80% gp91phox+ cells compared with healthy donor control subjects) long-term correction of X-CGD CD34+ cells.
慢病毒载体基因疗法用于治疗X连锁慢性肉芽肿病（X-CGD）已被证明是一种可行的方法，但转导细胞中随机的载体整合以及外源启动子产生的亚正常蛋白质表达，在长期安全性和有效性方面仍然令人担忧。先前基于基因组编辑的方法，使用化脓性链球菌Cas9 mRNA和寡脱氧核苷酸供体来修复基因突变，显示出恢复生理蛋白质表达的能力，但在静止的CD34+造血细胞中缺乏足够的效率以用于临床转化。在此，我们报告，短暂抑制p53结合蛋白1（53BP1）可显著增加（2.3倍）长期的同源定向修复，以实现对X-CGD CD34+细胞的高效（与健康供体对照受试者相比，80%的gp91phox+细胞）长期校正。
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