大阪大学Shimon Sakaguchi研究组取得一项新突破。他们揭示了人类T细胞全基因组CRISPR筛选揭示FOXP3调控因子。相关论文发表在2025年3月26日出版的《自然》杂志上。

在这里，该研究组进行了全基因组的CRISPR功能缺失筛选，编目了原代人T细胞中FOXP3诱导的基因调控决定因素，并在单细胞分辨率上通过Perturb-icCITE-seq表征了其作用。课题组发现RBPJ-NCOR抑制复合物是FOXP3表达的一种新的、环境特异性的负调控因子。靶向RBPJ的敲除增强了iT_reg的分化和功能，独立于典型的Notch信号。体外反复的细胞因子和T细胞受体信号刺激表明，与对照细胞相比，RBPJ缺陷的iT_reg细胞通过FOXP3增强子CNS2的DNA去甲基化，增强FOXP3的表达，表现出更高的表型稳定性。相反，RBPJ的过表达通过HDAC3直接调节FOXP3组蛋白乙酰化而有效抑制FOXP3的诱导。最后，在人源化motheme模型中，RBPJ消融的人iT_reg细胞比对照iT_reg细胞更有效地抑制异种移植物抗宿主病。总之，他们的发现揭示了FOXP3的新调控因子，并指出了提高自身免疫性疾病过继细胞治疗疗效的新途径。

研究人员表示，调节性T （T_reg）细胞特异性表达主转录因子FOXP3，在维持免疫耐受和体内平衡中起关键作用，并有可能彻底改变自身免疫性疾病的细胞治疗方法。虽然在TGFβ和IL-2的存在下刺激初始CD4⁺ T细胞可以在体外诱导FOXP3⁺ T_reg细胞（iT_reg细胞），但所产生的细胞通常不稳定，并且严重阻碍了翻译工作。了解T_reg分化调控网络的系统方法可能会导致更有效的基于T_reg细胞的治疗。

附：英文原文

Title: Genome-wide CRISPR screen in human T cells reveals regulators of FOXP3

Author: Chen, Kelvin Y., Kibayashi, Tatsuya, Giguelay, Ambre, Hata, Mayu, Nakajima, Shunsuke, Mikami, Norihisa, Takeshima, Yusuke, Ichiyama, Kenji, Omiya, Ryusuke, Ludwig, Leif S., Hattori, Kunihiro, Sakaguchi, Shimon

Issue&Volume: 2025-03-26

Abstract: Regulatory T (Treg) cells, which specifically express the master transcription factor FOXP3, have a pivotal role in maintaining immunological tolerance and homeostasis and have the potential to revolutionize cell therapies for autoimmune diseases1,2,3. Although stimulation of naive CD4+ T cells in the presence of TGFβ and IL-2 can induce FOXP3+ Treg cells in vitro (iTreg cells), the resulting cells are often unstable and have thus far hampered translational efforts4,5,6. A systematic approach towards understanding the regulatory networks that dictate Treg differentiation could lead to more effective iTreg cell-based therapies. Here we performed a genome-wide CRISPR loss-of-function screen to catalogue gene regulatory determinants of FOXP3 induction in primary human T cells and characterized their effects at single-cell resolution using Perturb-icCITE-seq. We identify the RBPJ–NCOR repressor complex as a novel, context-specific negative regulator of FOXP3 expression. RBPJ-targeted knockout enhanced iTreg differentiation and function, independent of canonical Notch signalling. Repeated cytokine and T cell receptor signalling stimulation in vitro revealed that RBPJ-deficient iTreg cells exhibit increased phenotypic stability compared with control cells through DNA demethylation of the FOXP3 enhancer CNS2, reinforcing FOXP3 expression. Conversely, overexpression of RBPJ potently suppressed FOXP3 induction through direct modulation of FOXP3 histone acetylation by HDAC3. Finally, RBPJ-ablated human iTreg cells more effectively suppressed xenogeneic graft-versus-host disease than control iTreg cells in a humanized mouse model. Together, our findings reveal novel regulators of FOXP3 and point towards new avenues to improve the efficacy of adoptive cell therapy for autoimmune disease.

DOI: 10.1038/s41586-025-08795-5

Source: https://www.nature.com/articles/s41586-025-08795-5