腺嘌呤碱基编辑恢复 CD3δ 重症联合免疫缺陷患者的人 T 细胞生成
Human T cell generation is restored in CD3δ severe combined immunodeficiency through adenine base editing.
机构信息
Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Department of Medicine, Division of Rheumatology, University of California, Los Angeles, Los Angeles, CA 90095, USA.
出版信息
Cell. 2023 Mar 30;186(7):1398-1416.e23. doi: 10.1016/j.cell.2023.02.027. Epub 2023 Mar 20.
CD3δ SCID is a devastating inborn error of immunity caused by mutations in CD3D, encoding the invariant CD3δ chain of the CD3/TCR complex necessary for normal thymopoiesis. We demonstrate an adenine base editing (ABE) strategy to restore CD3δ in autologous hematopoietic stem and progenitor cells (HSPCs). Delivery of mRNA encoding a laboratory-evolved ABE and guide RNA into a CD3δ SCID patient's HSPCs resulted in a 71.2% ± 7.85% (n = 3) correction of the pathogenic mutation. Edited HSPCs differentiated in artificial thymic organoids produced mature T cells exhibiting diverse TCR repertoires and TCR-dependent functions. Edited human HSPCs transplanted into immunodeficient mice showed 88% reversion of the CD3D defect in human CD34+ cells isolated from mouse bone marrow after 16 weeks, indicating correction of long-term repopulating HSCs. These findings demonstrate the preclinical efficacy of ABE in HSPCs for the treatment of CD3δ SCID, providing a foundation for the development of a one-time treatment for CD3δ SCID patients.
CD3δ SCID 是一种由 CD3D 基因突变引起的毁灭性先天性免疫缺陷，CD3D 编码 CD3/TCR 复合物的不变 CD3δ 链，对于正常的胸腺发生是必需的。我们展示了一种腺嘌呤碱基编辑 (ABE) 策略，用于恢复自体造血干细胞和祖细胞 (HSPCs) 中的 CD3δ。将编码实验室进化的 ABE 和指导 RNA 的 mRNA 递送到 CD3δ SCID 患者的 HSPCs 中，导致致病性突变的纠正率为 71.2%±7.85%（n=3）。编辑后的 HSPC 在人工胸腺类器官中分化，产生了具有多样化 TCR 库和 TCR 依赖性功能的成熟 T 细胞。编辑后的人类 HSPC 移植到免疫缺陷小鼠中，在 16 周后从小鼠骨髓中分离出的人类 CD34+细胞中，CD3D 缺陷的逆转率达到 88%，表明长期重编程 HSCs 的纠正。这些发现证明了 ABE 在 HSPCs 治疗 CD3δ SCID 中的临床前疗效，为开发一次性治疗 CD3δ SCID 患者的方法提供了基础。
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