Organoid-induced differentiation of conventional T cells from human pluripotent stem cells

Generation of T cells from pluripotent stem cells (PSC) has the potential to transform adoptive immunotherapy for cancer into universal donor, off-the-shelf cellular therapies. However, differentiation of human PSCs into fully mature T cells has been challenging with existing methods. We report that a 3D organoid method permitted efficient differentiation of human embryonic stem cell and induced pluripotent stem cell-derived mesoderm progenitors to mature, functional conventional T cells with a diverse T cell receptor (TCR) repertoire. This continuous culture system supported both hematopoietic induction and terminal differentiation to naïve, conventional CD3+CD8αβ+ and CD3+CD4+ T cells. Introduction of a Class I-restricted TCR in PSCs produced antigen-specific CD8αβ+ T cells lacking endogenous TCR expression. Functional assays and RNA sequencing aligned PSC-derived T cells with primary naïve conventional CD8+ T cells. The organoid system presented here provides an effective and scalable platform to generate functional mature T cells from human PSCs. Our goal was to analyze and compare transcriptome of human naïve T cells generated in Artificial Thymic Organoids (ATOs) from PSC with human naïve T cells from thymus and Cord blood ATOs.

多能干细胞（pluripotent stem cell, PSC）诱导生成T细胞，有望将癌症过继性免疫治疗变革为通用供体型、现成可用的细胞疗法。然而，现有技术难以将人类多能干细胞分化为完全成熟的T细胞。本研究报道，三维类器官方法可高效将人类胚胎干细胞及诱导多能干细胞来源的中胚层祖细胞，分化为具有多样化T细胞受体（T cell receptor, TCR）库的成熟功能性常规T细胞。该连续培养体系可同时支持造血诱导与终末分化，生成初始态常规CD3+CD8αβ+及CD3+CD4+ T细胞。在多能干细胞中导入I类限制性TCR，可获得内源性TCR表达缺失的抗原特异性CD8αβ+ T细胞。功能实验与RNA测序结果显示，多能干细胞诱导的T细胞与原发性初始态常规CD8+ T细胞具有高度一致性。本研究构建的类器官体系，可为人类多能干细胞诱导功能性成熟T细胞提供高效且可规模化的平台。本研究的目标为分析并比较：多能干细胞经人工胸腺类器官（Artificial Thymic Organoids, ATOs）诱导生成的人类初始T细胞，与来自胸腺及脐带血人工胸腺类器官的人类初始T细胞的转录组。