Sexual Dimorphism in T Cell Exhaustion and Bladder Cancer

项目1 -摘要 性别是一个未充分研究的生物学变量，对免疫功能有重大影响，包括在环境中 癌症。然而，在非癌症患者中观察到的性别偏见的免疫学机制， 生殖器官，如膀胱癌（BC），仍然难以捉摸。男性在乳腺癌发病率方面的偏见持续存在， 对其他已知的风险因素进行了调整，这表明了这种两性异形的基本生物学基础。 目的：明确雄激素、AR靶基因和性染色体- 编码的表观遗传修饰物导致肿瘤中CD 8 + T细胞耗竭的性别差异 微环境（TME）。影响：我们的研究将揭示免疫学基础和机制， CD 8 + T细胞耗竭导致BC中的性别偏见，对改善所有人的ICB方法具有意义 癌症有三个具体目标。目的1：确定AR-TCF 1和AR-TOX在CD 8 + T细胞耗竭中的作用 和肿瘤控制。雄激素影响T细胞并导致癌症的分子机制 进展尚不清楚。男性CD 8+肿瘤浸润淋巴细胞（TIL）优先采用TCF 1 + PE状态 由于AR直接激活Tcf 7。假说.雄激素信号传导建立了稳定的TCF 1驱动的 CD 8 + T细胞中的转录网络，有利于耗尽，通过调节TOX，另一个关键 转录因子（TF），其促进T细胞耗竭。目的2：确定AR-ZFP 148在T细胞中的作用 效应子分化TF ZFP 148调节非免疫细胞类型中的细胞增殖和存活， 还没有在T细胞生物学中研究过。单细胞RNA-seq分析表明Zfp 148表达偏向雄性 和AR刺激的PE CD 8 + T细胞中的调节活性。T细胞受体（TCR）参与和慢性 耗尽诱导Zfp 148在CD 8 + T细胞中表达，并且其缺失导致体外更好的效应子功能。的 Zfp 148无效T细胞的体内持久性受到损害。假说. AR调节的ZFP 148可以抑制 CD 8 + T细胞效应子功能，并有助于男性偏好的BC。目的3：研究AR的作用， 表观遗传修饰剂在CD 8 + T细胞耗竭和肿瘤对免疫疗法的反应中的作用。性的影响 BC中染色体编码的表观遗传修饰物-包括KDM 5D，UTY和KDM 6A-对免疫力的影响 特别是在CD 8+区室中的免疫调节是该领域的主要空白。在与项目2的合作中，我们发现， Y染色体（LOY）（尤其是基因Kdm 5d和Uty）导致BC中的CD 8 + T细胞耗竭。 假说.性激素和性染色体编码的表观遗传修饰物有助于CD 8 + T细胞 衰竭和BC对ICB的敏感性。

PROJECT 1 – SUMMARY Sex is an understudied biological variable with significant influence on immune function, including in the context of cancer. However, the immunological mechanisms underlying the sex bias seen in cancers from non- reproductive organs, such as bladder cancer (BC), remain elusive. Male bias in BC incidence persists after adjustment for other known risk factors suggesting a fundamental biological basis for this sexual dimorphism. Objective: To define the molecular mechanisms by which androgens, AR target genes, and sex chromosome- encoded epigenetic modifiers contribute to sex differences in CD8+ T cell exhaustion in the tumor microenvironment (TME). Impact: Our study will uncover an immunological basis and mechanism by which CD8+ T cell exhaustion contributes to sex bias in BC, with implications for improving ICB approaches for all cancer via three specific aims. Aim 1: Determine the role of AR-TCF1 and AR-TOX in CD8+ T cell exhaustion and tumor control. The molecular mechanisms by which androgens influence T cells and contribute to cancer progression are unclear. Male CD8+ tumor-infiltrating lymphocytes (TILs) preferentially adopt a TCF1+ PE state due to direct activation of Tcf7 by AR. Hypothesis. Androgen signaling establishes a stable TCF1-driven transcriptional network in CD8+ T cells that favors exhaustion, through the modulation of TOX, another critical transcription factor (TF) that promotes T cell exhaustion. Aim 2: Determine the role of AR-ZFP148 in T cell effector differentiation. TF ZFP148 regulates cell proliferation and survival in non-immune cell types but has not been studied in T cell biology. Single-cell RNA-seq analyses demonstrated male-biased Zfp148 expression and regulatory activity in PE CD8+ T cells stimulated by AR. T cell receptor (TCR) engagement and chronic exhaustion induced Zfp148 expression in CD8+ T cells, and its deletion led to better effector function in vitro. The in vivo persistence of Zfp148 null T cells was compromised. Hypothesis. AR-regulated ZFP148 can suppress CD8+ T cell effector function and contributes to male-biased BC. Aim 3: Investigate the role of AR and epigenetic modifiers in CD8+ T cell exhaustion and tumor response to immunotherapy. The impact of sex chromosome-encoded epigenetic modifiers in BC – including KDM5D, UTY, and KDM6A – on immunity especially in the CD8+ compartment is a major gap in the field. In collaboration with Project 2, we found that loss of the Y chromosome (LOY) (especially the genes Kdm5d and Uty) contributes to CD8+ T cell exhaustion in BC. Hypothesis. Sex hormones and sex chromosome-encoded epigenetic modifiers contribute to CD8+ T cell exhaustion and BC sensitivity to ICB.