Istituto di Ricerche Farmacologiche Mario Negri IRCCS, 24126 Bergamo, Italy.
Int J Mol Sci. 2023 Jul 22;24(14):11810. doi: 10.3390/ijms241411810.
Induced pluripotent stem cells (iPSC) have huge potential as cell therapy for various diseases, given their potential for unlimited self-renewal and capability to differentiate into a wide range of cell types. Although autologous iPSCs represents the ideal source for patient-tailored regenerative medicine, the high costs of the extensive and time-consuming production process and the impracticability for treating acute conditions hinder their use for broad applications. An allogeneic iPSC-based strategy may overcome these issues, but it carries the risk of triggering an immune response. So far, several approaches based on genome-editing techniques to silence human leukocyte antigen class I (HLA-I) or II (HLA-II) expression have been explored to overcome the immune rejection of allogeneic iPSCs. In this study, we employed the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9) system to delete the β2-Microglobulin () and the Class II Major Histocompatibility Complex Transactivator () genes, essential for the correct surface expression of HLA-I and HLA-II proteins. The resulting hypoimmunogenic iPSC line has a normal karyotype, expresses the pluripotency stem cell markers, and is capable of differentiating into the three embryonic germ layers. Furthermore, we showed that it specifically retains the ability to differentiate towards different liver cells, such as endothelial-like cells, hepatocyte-like cells, and hepatic stellate-like cells. Our results indicate that hypoimmunogenic iPSCs could give a new cost-effective and off-the-shelf opportunity for cell therapy in liver diseases.
诱导多能干细胞（iPSC）具有巨大的潜力作为细胞治疗各种疾病，因为它们具有无限自我更新的潜力，并能够分化为广泛的细胞类型。虽然自体 iPSC 是为患者量身定制再生医学的理想来源，但广泛而耗时的生产过程的高成本以及治疗急性疾病的不切实际性阻碍了它们的广泛应用。同种异体 iPSC 为基础的策略可能克服这些问题，但它具有引发免疫反应的风险。到目前为止，已经探索了几种基于基因组编辑技术的方法来沉默人类白细胞抗原 I 类（HLA-I）或 II 类（HLA-II）的表达，以克服同种异体 iPSC 的免疫排斥。在这项研究中，我们使用了 CRISPR/Cas9（簇状规律间隔短回文重复/CRISPR 相关蛋白 9）系统来删除β2-微球蛋白（β2-Microglobulin）和 II 类主要组织相容性复合物转录激活物（Class II Major Histocompatibility Complex Transactivator）基因，这些基因对于 HLA-I 和 HLA-II 蛋白的正确表面表达是必不可少的。由此产生的低免疫原性 iPSC 系具有正常的核型，表达多能干细胞标志物，并能够分化为三个胚胎 germ 层。此外，我们表明它特别保留了向不同的肝细胞分化的能力，如内皮样细胞、肝细胞样细胞和肝星状样细胞。我们的结果表明，低免疫原性 iPSC 可为肝脏疾病的细胞治疗提供一种新的具有成本效益的现成机会。
