PDF(Pubmed)
Abstract：
Robust allogeneic immune reactions after transplantation impede the translational pace of human embryonic stem cells (hESCs)-based therapies. Selective genetic editing of human leucocyte antigen (HLA) molecules has been proposed to generate hESCs with immunocompatibility, which, however, has not been specifically designed for the Chinese population yet. Herein, we explored the possibility of customizing immunocompatible hESCs based on Chinese HLA typing characteristics. We generated an immunocompatible hESC line by disrupting HLA-B, HLA-C, and CIITA genes while retaining HLA-A*11:01 (HLA-A*11:01-retained, HLA-A11R ), which covers ~21% of the Chinese population. The immunocompatibility of HLA-A11R hESCs was verified by in vitro co-culture and confirmed in humanized mice with established human immunity. Moreover, we precisely knocked an inducible caspase-9 suicide cassette into HLA-A11R hESCs (iC9-HLA-A11R ) to promote safety. Compared with wide-type hESCs, HLA-A11R hESC-derived endothelial cells elicited much weaker immune responses to human HLA-A11+ T cells, while maintaining HLA-I molecule-mediated inhibitory signals to natural killer (NK) cells. Additionally, iC9-HLA-A11R hESCs could be induced to undergo apoptosis efficiently by AP1903. Both cell lines displayed genomic integrity and low risks of off-target effects. In conclusion, we customized a pilot immunocompatible hESC cell line based on Chinese HLA typing characteristics with safety insurance. This approach provides a basis for establishment of a universal HLA-AR bank of hESCs covering broad populations worldwide and may speed up the clinical application of hESC-based therapies.
摘要：
移植后强烈的同种异体免疫反应阻碍了基于人胚胎干细胞（hESC）的疗法的转化速度。人类白细胞抗原（HLA）分子的选择性遗传编辑已被提出来产生具有免疫相容性的hESCs，which,然而,还没有专门为中国人设计。在这里，我们探索了根据中国人HLA分型特征定制免疫相容性hESCs的可能性.我们通过破坏HLA-B产生了免疫相容的hESC系，HLA-C,和CIITA基因，同时保留HLA-A*11:01(HLA-A*11:01保留，HLA-A11R),占中国人口的21%。通过体外共培养验证了HLA-A11RhESC的免疫相容性，并在建立了人类免疫力的人源化小鼠中得到了证实。此外，我们将一个诱导型caspase-9自杀盒精确地敲入HLA-A11RhESCs(iC9-HLA-A11R)以提高安全性.与宽型hESC相比，HLA-A11RhESC来源的内皮细胞对人类HLA-A11+T细胞的免疫反应较弱，同时维持HLA-I分子介导的对自然杀伤（NK）细胞的抑制信号。此外,AP1903可有效诱导iC9-HLA-A11RhESC发生凋亡。两种细胞系均显示基因组完整性和脱靶效应的低风险。总之,我们根据中国人HLA分型特征定制了具有安全性保险的飞行员免疫相容性hESC细胞系。这种方法为建立覆盖全球广泛人群的hESC通用HLA-AR库提供了基础，并可能加快基于hESC的疗法的临床应用。
