Androgen Activation of Innate Immune Signaling to Enhance Prostate Cancer Immune Response

双极雄激素疗法(BAT)是治疗去势抵抗前列腺癌的一种矛盾的方法。 (CRPC)，睾丸激素水平在精液上清液和去势精液浓度之间快速循环。 了解BAT在分子和细胞水平上的工作原理可能有助于将BAT与 其他药物可达到提高疗效和肿瘤反应的目的。之前的观察表明， 超生理学睾酮(SupraT)诱导DNA双链断裂(DSB)。据推测，如果 如果不加以修复，DSB可能会导致细胞危机和细胞凋亡。在这项提案中，我们提供了新的证据 由Suprats诱导的未修复的双链断裂被送到自噬小体，在那里它们激活胞质核 触发下游干扰素刺激基因(ISG)和先天免疫途径的酸传感器。基座 根据我们的初步数据，我们提出了一个新的想法：i)由Suprats引起的未修复的DSB被路由到 特殊的自噬降解称为吞核作用；ii)Suprats诱导的自噬DNA可以激活 细胞质DNA传感通路~特别是核酸传感通路(cGAS-STING和RIG-I 途径)；iii)在前列腺癌中，由Suprats激活的核酸传感途径将更加明显 有DNA修复缺陷的；以及iv)BAT可能特别地激活先天和获得性免疫细胞，在 有DNA修复缺陷的患者。在这项提案中，我们将确定核酸传感器在调节 前列腺癌中Suprats的免疫信号转导。利用接受BAT治疗的前列腺癌患者的肿瘤活检组织，我们将评估 核酸感受器介导的先天免疫信号是否作为治疗的分子决定因素 回应。为了验证我们的假设，我们将使用一些创新的工具和资源，包括CRISPR-CAS9 生成基因敲除细胞模型，GeoMx数字空间分析肿瘤中的免疫格局 微环境，独特的前列腺癌患者来源的异种移植模型，人源化的小鼠模型 功能先天和获得性免疫细胞，以及接受BAT治疗的患者的血清和肿瘤活检。我们认为 这些独特的资源使我们能够很好地承担拟议的工作，并立即产生临床影响。我们有 组建了一支基础和临床前列腺癌生物学和免疫疗法专家团队，他们将为 独一无二的专业知识，成功完成我们的目标。成功完成拟议工作将产生以下成果： A)对Suprats调节免疫反应的机械性见解，b)对激活的宝贵临床见解 BAT对免疫细胞的作用，c)基于肿瘤和血清的新标记物，可用于开发 预测治疗反应的生物标志物，以及d)为战略性地利用BAT激活提供了理由 免疫反应，并将其与增强适应性免疫系统的免疫疗法相结合，例如 免疫关卡阻断和T细胞治疗达到协同作用。

Bipolar androgen therapy (BAT) is a paradoxical approach for the treatment of castration-resistant prostate cancer (CRPC) whereby testosterone levels are rapidly cycled between supraphysiologic and castrate concentrations. Understanding how BAT works at the molecular and cellular levels might help in rationally combining BAT with other agents to achieve increased efficacy and tumor responses. Previous observations suggest that supraphysiological testosterone (SupraT) induce DNA double strand breaks (DSB). It has been speculated that if left unrepaired, DSBs may lead to cellular crisis and apoptosis. In this proposal, we provide novel evidence that unrepaired DSBs induced by SupraTs are routed to the autophagosomes where they activate cytoplasmic nucleic acid sensors that trigger the downstream interferon stimulated genes (ISGs) and innate immune pathways. Based on our preliminary data, we propose a novel idea that: i) Unrepaired DSBs induced by SupraTs are routed for specialized autophagic degradation termed nucleophagy; ii) SupraTs induced autophagosomal DNA can activate cytoplasmic DNA sensing pathways~ specifically the nucleic acid sensing pathway (cGAS-STING and RIG-I pathway); iii) Activation of nucleic acid sensing pathway by SupraTs would be more pronounced in prostate tumors that have DNA repair defect; and iv) BAT might activate innate and adaptive immune cells specially, in a subset of patients having DNA repair defect. In this proposal, we will determine the role of nucleic acid sensors in mediating immune signaling by SupraTs in PCa. Utilizing tumor biopsies from PCa patients receiving BAT, we will evaluate whether nucleic acid sensor mediated innate immune signaling serves as a molecular determinant of treatment response. To test our hypothesis, we will utilize several innovative tools and resources including CRISPR-Cas9 generated knockout cellular models, GeoMx Digital Spatial profiling of immune landscape in the tumor microenvironment, unique patient derived prostate cancer xenografts models, a humanized mouse models that has functional innate and adaptive immune cells, and serum and tumor biopsies from patients receiving BAT. We think these unique resources position us well to undertake the proposed work with immediate clinical impact. We have assembled a team of experts in basic and clinical prostate cancer biology and immunotherapy who will provide their unique expertise to successfully accomplish our goals. Successful completion of the proposed work will generate: a) mechanistic insights into modulation of immune response by SupraTs, b) valuable clinical insights into activation of immune cells by BAT, c) novel tumor- and serum-based markers that can be utilized for the development of biomarkers predictive of therapy response, and d) provide a rationale for strategically utilizing BAT to activate immune response and combine it with immunotherapeutics that empower the adaptive immune system such as immune checkpoint blockade and T-cell therapeutics to achieve synergy.