Xu Saijuan, Liang Dan, Wang Qiudao, Cheng Yan, Xie Da, Gui Yang, Zhang Haokun, Feng Changrui, Zhao Feiyan, Ren Wendan, Sun Gongrui, Yang Yang, Li Lin, Lai Yongrong, Fu Bin, Lu Yuming, Wang Zi Jun, Wu Yuxuan
Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China.
Department of Obstetrics and Gynecology, the First Affiliated Hospital of Anhui Medical University, Hefei, China.
Nat Biomed Eng. 2025 Aug 12. doi: 10.1038/s41551-025-01480-y.
Ex vivo autologous haematopoietic stem cell (HSC) gene therapy provides a promising treatment option for haematological disorders. However, current methods involve complex processes and chemotherapeutic conditioning, leading to limited accessibility for treatment and major side effects. Here we develop antibody-free targeted lipid nanoparticles (LNPs) for mRNA delivery to HSCs in vivo, enabling efficient base editing of the γ-globin gene (HBG1/2) promoter target in human HSCs to reactivate fetal haemoglobin in derived erythroid cells. Delivery of ABE8e/sgRNA mRNA with optimized LNPs achieves efficient in vivo base editing of HBG1/2 in transfusion-dependent β-thalassaemia (TDT) patient-derived HSCs engrafted in immunodeficient NCG-X mice, showing restored globin chain balance in erythroid cells. Our research indicates that using LNPs for genome editor delivery achieves efficient editing of endogenous genes of human HSCs. This non-viral delivery system eliminates the need for collecting or mobilizing HSCs, providing a potent and one-time treatment potential for blood disorders such as sickle cell disease and TDT.
体外自体造血干细胞（HSC）基因治疗为血液系统疾病提供了一种有前景的治疗选择。然而，目前的方法涉及复杂的过程和化疗预处理，导致治疗的可及性有限且有严重的副作用。在此，我们开发了无抗体靶向脂质纳米颗粒（LNPs），用于在体内将mRNA递送至造血干细胞，从而能够对人类造血干细胞中的γ-珠蛋白基因（HBG1/2）启动子靶点进行高效碱基编辑，以重新激活衍生红细胞中的胎儿血红蛋白。用优化的LNPs递送ABE8e/sgRNA mRNA可在移植到免疫缺陷NCG-X小鼠体内的输血依赖型β地中海贫血（TDT）患者来源的造血干细胞中实现对HBG1/2的高效体内碱基编辑，显示出红细胞中珠蛋白链平衡的恢复。我们的研究表明，使用LNPs递送基因组编辑器可实现对人类造血干细胞内源基因的高效编辑。这种非病毒递送系统无需收集或动员造血干细胞，为镰状细胞病和TDT等血液疾病提供了一种强效且一次性的治疗潜力。
