患者源性肝細胞的高效擴增和Crispr-Cas9介導的基因校正治療遺傳性肝病

2024年5月，1中國科學院上海生物化學與細胞生物學研究所多細胞系統重點實驗室，中國科學院分子細胞科學**研究中心，2上海交通大學醫學院仁濟醫院肝臟外科，華東師范大學生物醫學科學研究所，生命科學學院，3上海市調控生物學重點實驗室，上海市基因組編輯與細胞治療前沿科學中心，華東師范大學，4 上海工業大學人類研究所，5北京大學生命科學學院，細胞增殖與分化教育部重點實驗室，北京大學基因組編輯研究中心;6中國科學院大學，7上海工業大學生命科學與技術學院（1Key Laboratory of Multi-Cell Systems, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science,Chinese Academy of Sciences,

2Department of Liver Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University,

3Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute ofBiomedical Sciences and School of Life Sciences, East China Normal University,

4iHuman Institute, ShanghaiTech University,

5The MOE Key Laboratory of Cell Proliferation and Differentiation, School of Life Sciences, Genome Editing Research Center, Peking University,

6University of Chinese Academy of   Sciences,

7School of Life Science and Technology, ShanghaiTech University,）Qiang Xia

and Lijian Hui研究團隊在《Cell Stem Cell》上發表論文：

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

“患者源性肝細胞的高效擴增和Crispr-Cas9介導的基因校正治療遺傳性肝病”

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.

摘要：

在實體器官，特別是肝臟中進行基于細胞的體外基因治療，在技術上具有挑戰性。在這里，我們報告了肝細胞治療臨床應用的可行策略。我們首先通過大規模擴增患者源性肝細胞獲得了高質量的自體肝細胞。此外，增殖的患者源性肝細胞與通過篩選鑒定的AAV2.7m8變體一起，有效地實現了Crispr-Cas9介導的靶向整合，實現了FAH或OTC致病性突變的功能糾正。重要的是，這些經過編輯的肝細胞以高水平重新填充受傷小鼠的肝臟并經歷成熟，成功地治療了移植后酪氨酸血癥小鼠。我們的研究結合了患者來源的可移植肝細胞的體外大規模細胞擴增和基因編輯，具有治療人類肝臟疾病的潛力。

該論文中，L-Wnt3a細胞系的體外培養是使用Ausbian特級胎牛血清完成的。欲了解或購買Ausbian特級胎牛血清可以聯系北京締一生物400-166-8600.