人类调节性T细胞（Tregs）是组织修复、自身免疫性疾病和癌症的关键调节因子。然而，在癌症治疗中，抑制Tregs的抑制功能而不影响免疫稳态是具有挑战性的。鉴定可能区分肿瘤特异性Tregs的路径对于研究至关重要，然而，控制肿瘤内Treg基因表达的转录程序，以及与健康组织中的Tregs不同的转录程序，仍然大部分未知。我们对头颈部鳞状细胞癌（HNSCC）患者肿瘤和外周血液中的CD4+ T细胞以及健康供体扁桃体组织和外周血液中的CD4+ T细胞进行了单细胞转录组学分析。我们发现在肿瘤微环境（TME）中高度富集了表达多个肿瘤坏死因子受体（TNFR）基因（TNFR+ Tregs）的活化Tregs亚群，与非肿瘤组织和外周血液相比。TNFR+ Tregs与HNSCC以及多种实体肿瘤类型的恶化预后有关。机制上，转录因子BATF是一个基因调控网络的核心组成部分，该网络控制TNFR+ Tregs的关键方面。通过CRISPR-Cas9介导的BATF基因敲除实验、大规模RNA测序、免疫表型分析和体外功能实验，我们证实了BATF在限制过度活化和促进人类活化Tregs存活中的中心作用。最后，我们鉴定了与BATF驱动的转录网络在HNSCC患者的肿瘤内Tregs中相关的一组表面分子。这些发现揭示了高度抑制性肿瘤内Tregs的主要转录调控因子，突显了在癌症治疗中干预的潜在机会，而不影响免疫稳态。

Human regulatory T cells (Tregs) are crucial regulators of tissue repair, autoimmune diseases, and cancer. However, it is challenging to inhibit the suppressive function of Tregs for cancer therapy without affecting immune homeostasis. Identifying pathways that may distinguish tumor-restricted Tregs is important, yet the transcriptional programs that control intratumoral Treg gene expression, and that are distinct from Tregs in healthy tissues, remain largely unknown. We profiled single-cell transcriptomes of CD4+ T cells in tumors and peripheral blood from patients with head and neck squamous cell carcinomas (HNSCC) and those in nontumor tonsil tissues and peripheral blood from healthy donors. We identified a subpopulation of activated Tregs expressing multiple tumor necrosis factor receptor (TNFR) genes (TNFR+ Tregs) that is highly enriched in the tumor microenvironment (TME) compared with nontumor tissue and the periphery. TNFR+ Tregs are associated with worse prognosis in HNSCC and across multiple solid tumor types. Mechanistically, the transcription factor BATF is a central component of a gene regulatory network that governs key aspects of TNFR+ Tregs. CRISPR-Cas9-mediated BATF knockout in human activated Tregs in conjunction with bulk RNA sequencing, immunophenotyping, and in vitro functional assays corroborated the central role of BATF in limiting excessive activation and promoting the survival of human activated Tregs. Last, we identified a suite of surface molecules reflective of the BATF-driven transcriptional network on intratumoral Tregs in patients with HNSCC. These findings uncover a primary transcriptional regulator of highly suppressive intratumoral Tregs, highlighting potential opportunities for therapeutic intervention in cancer without affecting immune homeostasis.