Elucidating mechanisms of T cell development can guide in vitro T cell differentiation from induced pluripotent stem cells (iPSCs) and facilitate off-the-shelf T cell-based immunotherapies. Using a stroma-free human iPSC-T cell differentiation platform, we screened for epigenetic modulators that influence T cell specification and identified the H3K9-directed histone methyltransferases G9a/GLP as repressors of T cell fate. We show that G9a/GLP inhibition during specific time windows of differentiation of hematopoietic stem and progenitor cells (HSPCs) skews cell fates toward lymphoid lineages. Inhibition of G9a/GLP promotes the production of lymphoid cells during zebrafish embryonic hematopoiesis, demonstrating the evolutionary conservation of G9a/GLP function. Importantly, chemical inhibition of G9a/GLP facilitates the generation of mature iPSC-T cells that bear transcriptional similarity to peripheral blood αβ T cells. When engineered to express chimeric antigen receptors, the epigenetically engineered iPSC-T cells exhibit enhanced effector functions in vitro and durable, persistent antitumor activity in a xenograft tumor-rechallenge model.

中文翻译：

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通过化学抑制 G9a/GLP 实现源自人多能干细胞的 CAR T 细胞的成熟和持久性


阐明 T 细胞发育的机制可以指导 体外 T 细胞与诱导多能干细胞 （iPSC） 的分化，并促进现成的基于 T 细胞的免疫疗法。使用无基质的人 iPSC-T 细胞分化平台，我们筛选了影响 T 细胞规格的表观遗传调节剂，并确定了 H3K9 导向的组蛋白甲基转移酶 G9a/GLP 作为 T 细胞命运的抑制因子。我们表明，在造血干细胞和祖细胞 （HSPCs） 分化的特定时间窗口内，G9a/GLP 抑制使细胞命运偏向于淋巴谱系。抑制 G9a/GLP 促进斑马鱼胚胎造血过程中淋巴细胞的产生，证明 G9a/GLP 功能的进化保守性。重要的是，G9a/GLP 的化学抑制促进了成熟 iPSC-T 细胞的产生，这些细胞与外周血 αβ T 细胞具有转录相似性。当经过基因改造以表达嵌合抗原受体时，表观遗传工程改造的 iPSC-T 细胞在 体外表现出增强的效应功能，并在异种移植肿瘤再攻击模型中表现出持久、持久的抗肿瘤活性。