肿瘤细胞治疗研究
英文原题：An integrated SOX4-driven transcriptional and metabolic program governs tumor-infiltrating regulatory T cells in ovarian cancer.
SOX4是TI-Treg抑制功能和代谢适应性的核心调控因子，是OC一个有前景的治疗靶点。
调节性T细胞（Tregs）在卵巢癌（OC）中抑制抗肿瘤免疫，是免疫治疗的有前景靶点。然而，肿瘤浸润Tregs（TI-Tregs）的异质性和调控机制仍定义不清。在此，我们旨在描绘TI-Treg程序，以识别潜在的治疗靶点。
来自OC、癌旁组织和外周血的CD4⁺CD25⁺CD127⁻ Tregs通过单细胞RNA测序和空间转录组学进行了分析，并使用SCENIC推断了调控网络。SOX4的功能和机制研究通过缺氧/肿瘤条件模型、CRISPR-Cas9扰动、异位过表达、Cut&Tag分析和氧化磷酸化（OXPHOS）抑制进行。
我们鉴定出九个转录上不同的Treg亚群，揭示了TI-Tregs中高度活化和免疫抑制的状态。这些TI-Tregs表现出TNFRSF4、TNFRSF9、TNFRSF18和CTLA4的强共表达，其瘤内丰度增加与较差的总生存期独立相关。SCENIC分析确定SOX4是定义TI-Treg身份的首要调控子，空间转录组学揭示SOX4⁺ Tregs在肿瘤边缘形成免疫调节屏障。在成熟的小鼠OC模型中，这种表型身份由缺氧微环境和TCR刺激稳健诱导。在机制上，SOX4反式激活MT1X以促进线粒体适应性和OXPHOS。一致地，Cut&Tag分析显示SOX4敲低后OXPHOS相关位点的染色质可及性降低。此外，CRISPR-Cas9介导的SOX4破坏降低了FOXP3表达和其他抑制性标志物，而SOX4过表达以OXPHOS依赖的方式增强了FOXP3表达。值得注意的是，OXPHOS的药理学抑制消除了这一效应。
BACKGROUND: Regulatory T cells (Tregs) suppress antitumor immunity in ovarian cancer (OC) and are promising targets for immunotherapy. However, the heterogeneity and regulatory mechanisms of tumor-infiltrating Tregs (TI-Tregs) remain poorly defined. Here, we aim to delineate TI-Treg programs to identify potential therapeutic targets. METHODS: CD4⁺CD25⁺CD127⁻ Tregs from OC, adjacent tissues, and peripheral blood were profiled by single-cell RNA sequencing and spatial transcriptomics, with regulatory networks inferred using SCENIC. Functional and mechanistic studies of SOX4 were performed using hypoxic/tumor-conditioned models, CRISPR-Cas9 perturbation, ectopic overexpression, Cut&Tag profiling, and oxidative phosphorylation (OXPHOS) inhibition. RESULTS: We identified nine transcriptionally distinct Treg subsets, revealing a highly activated and immunosuppressive state among TI-Tregs. These TI-Tregs exhibited strong co-expression of TNFRSF4 , TNFRSF9 , TNFRSF18 , and CTLA4 , and their increased intratumoral abundance was independently associated with poorer overall survival. SCENIC analysis identified SOX4 as the top regulon defining TI-Treg identity, with spatial transcriptomics revealing SOX4⁺ Tregs forming an immunoregulatory barrier at tumor margins. This phenotypic identity was robustly induced by the hypoxic microenvironment and TCR stimulation in a well-established murine OC model. Mechanistically, SOX4 transactivated MT1X to promote mitochondrial fitness and OXPHOS. Consistently, Cut&Tag profiling showed reduced chromatin accessibility at OXPHOS-related loci following SOX4 depletion. Furthermore, CRISPR-Cas9-mediated disruption of SOX4 reduced FOXP3 expression and other suppressive markers, whereas SOX4 overexpression enhanced FOXP3 expression in an OXPHOS-dependent manner. Notably, pharmacological inhibition of OXPHOS abolished this effect. CONCLUSIONS: SOX4 is a central regulator of TI-Treg suppressive function and metabolic fitness, representing a promising therapeutic target for OC.
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