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

• DOI: 10.1016/j.stem.2024.04.022
• Publication Date: 2024-05-20T14:31:01Z
• Authors: 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
• Keywords: Ex vivo; Tyrosinemia

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