中国科学院分子细胞科学卓越创新中心惠利健小组取得一项新突破。他们报道了患者源性肝细胞的高效扩增和CRISPR-Cas9介导的基因校正治疗遗传性肝病。相关论文于2024年5月20日发表于国际顶尖学术期刊《细胞—干细胞》杂志上。

据悉，在实体器官，特别是肝脏中进行基于细胞的体外基因治疗，在技术上具有挑战性。

课题组报告了肝细胞治疗临床应用的可行策略。研究组首先通过大规模扩增患者源性肝细胞，生成了高质量的自体肝细胞。此外，增殖的患者源性肝细胞与通过筛选鉴定的AAV2.7m8变体一起，有效地实现了CRISPR-Cas9介导的靶向整合，实现了FAH或OTC致病性突变的功能纠正。

重要的是，这些编辑过的肝细胞以高水平重新填充损伤的小鼠肝脏，并经历成熟，成功地治疗了小鼠移植后酪氨酸血症。他们的研究结合了患者来源的可移植肝细胞的体外大规模细胞扩增和基因编辑，具有治疗人类肝脏疾病的潜力。

附：英文原文

Title: Efficient expansion and CRISPR-Cas9-mediated gene correction of patient-derived hepatocytes for treatment of inherited liver diseases

Author: Kun Zhang, Ping Wan, Liren Wang, Zhen Wang, Fangzhi Tan, Jie Li, Xiaolong Ma, Jin Cen, Xiang Yuan, Yang Liu, Zhen Sun, Xi Cheng, Yuanhua Liu, Xuhao Liu, Jiazhi Hu, Guisheng Zhong, Dali Li, Qiang Xia, Lijian Hui

Issue&Volume: 2024-05-20

Abstract: Cell-based ex vivo gene therapy in solid organs, especially the liver, has proven technically challenging. Here, we report a feasible strategy for the clinical application of hepatocyte therapy. We first generated high-quality autologous hepatocytes through the large-scale expansion of patient-derived hepatocytes. Moreover, the proliferating patient-derived hepatocytes, together with the AAV2.7m8 variant identified through screening, enabled CRISPR-Cas9-mediated targeted integration efficiently, achieving functional correction of pathogenic mutations in FAH or OTC. Importantly, these edited hepatocytes repopulated the injured mouse liver at high repopulation levels and underwent maturation, successfully treating mice with tyrosinemia following transplantation. Our study combines ex vivo large-scale cell expansion and gene editing in patient-derived transplantable hepatocytes, which holds potential for treating human liver diseases.

DOI: 10.1016/j.stem.2024.04.022

Source: https://www.cell.com/cell-stem-cell/fulltext/S1934-5909(24)00177-2