Targeting Non-Canonical STING Signaling to Treat SPOP Mutant Castration-Resistant Prostate Cancer

靶向非经典 STING 信号传导治疗 SPOP 突变去势抵抗性前列腺癌

• 批准号: 10709358
• 负责人: Timothy Charles Thompson
• 金额: $ 66.83万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10709358
• 关键词: Aftercare; Androgens; Apoptotic; Automobile Driving; Binding Proteins; Biological Markers; Blood specimen; CHD1 gene; Castrate sensitive prostate cancer; Castration; Cell Survival; Cell model; Cells; Chromatin Remodeling Factor; Clinical; Clinical Data; Clinical Markers; Clinical Trials; Coculture Techniques; Combined Modality Therapy; Consensus; Cullin Proteins; DNA Damage; DNA Sequence Alteration; DNA analysis; Data Set; Epithelium; Equilibrium; Evaluation; Genes; Genetic; Growth; Heterozygote; High Mobility Group Proteins; Human; Immune signaling; Interferon-beta; Interleukin-6; Macrophage; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Methods; Missense Mutation; Modeling; Mus; Mutation; Other Genetics; Pathway interactions; Patients; Phenotype; Phosphorylation; Poly(ADP-ribose) Polymerase Inhibitor; Proteins; Proteomics; Regulation; Resistance; Role; STAT3 gene; Sampling; Signal Transduction; Stat3 protein; Stimulator of Interferon Genes; T-Lymphocyte; Testing; The Cancer Genome Atlas; Therapeutic Effect; Tissue Sample; Treatment Efficacy; Tumor Suppressor Genes; Up-Regulation; castration resistant prostate cancer; cell growth; chromatin protein; cytokine; experimental study; genetic signature; in vivo; mutant; neoplastic cell; neutralizing antibody; paracrine; patient biomarkers; patient subsets; prostate cancer cell; prostate cancer model; recruit; resistance mechanism; response; stable isotope; targeted treatment; therapeutic evaluation; tumor; tumor growth; tumor microenvironment; ubiquitin ligase

PROJECT SUMMARY/ABSTRACT Tumor suppressor gene speckle-type POZ protein (SPOP), a substrate adaptor of cullin3-RING ubiquitin ligase, demonstrates heterozygous missense mutations in up to 15% of prostate cancers, yet the functional significance of these SPOP mutations is largely unknown. We identified SPOP-binding consensus motifs in multiple proteins of the canonical cGAS-STING and non-canonical STING-NF-κB pathways and demonstrated that human and mouse STING protein is a bona fide SPOP target. Analysis of SPOP mutant (SPOPmut) and SPOP wild-type (SPOPwt) castration-resistant prostate cancer (CRPC) clinical data sets revealed a 29-gene “SPOPmut gene signature”, which reflected STING-NF-κB signaling activity and suggested a role for the recruitment of tumor microenvironment (TME)–facilitated tumor cell growth and survival in SPOPmut CRPC. Further evaluation of the “SPOPmut gene signature” in primary, untreated prostate cancer from TCGA revealed a subset of SPOP mutant tumors, as well as a subset of CHD1 mutant (CHD1mut) tumors, that are enriched for the signature. Importantly, co-mutations in SPOP and CHD1 (chromatin remodeling factor) are well documented. In stably transduced human and mouse SPOPmut (SPOPF102C and SPOPF133V)–expressing CRPC models, we demonstrated upregulation of non-canonical STING-NF-κB-IL-6 pathway proteins, STAT3, and HMG proteins involved in promoting secretory pathway activities. We also showed that PARP inhibitor (PARPi) treatment of SPOPmut CRPC cells induces DNA damage, activates canonical cGAS-STING-TBK-IFN-ß signaling and suppresses non- canonical STING-NF-κB-IL-6/STAT3, in part through inhibitory phosphorylation (S754-STAT3), leading to growth suppression and apoptotic signaling. Olaparib (OLA) increased IFN-β secretion and reduced viability to a greater extent in SPOPmut prostate cancer cells than in control cells in coculture with macrophages versus monoculture. In addition, activation of cGAS-STING and induction of IFN-ß in macrophages were demonstrated only in OLA- treated coculture models, and neutralizing antibody experiments showed that paracrine regulation of OLA- mediated growth suppression involved IFN-β induction and IL-6 suppression of SPOPmut prostate cancer cells. We hypothesize that SPOPmut, CHD1 deletion (CHD1del) and SPOP + CHD1 co-mutations in prostate cancers promote PARPi or PARPi + anti-IL-6 therapeutic efficacy through enhanced synthetic lethality driven by increased, unrepaired DNA damage, which leads to a shift in the balance toward canonical cGAS-STING-IFN-ß signaling and suppression of IL-6/STAT3. We propose to analyze the underlying mechanisms of the SPOPmut and CHD1del prostate cancer phenotype and define these genetically driven TME alterations and their protein effectors through macrophage reprogramming using state-of-the-art proteomics (Aim 1); analyze the efficacy of PARPi and anti-IL-6 treatment in SPOPmut (F133V), CHD1del, and SPOPmut;CHD1del RM-1-BM syngeneic CRPC models (Aim 2); and analyze clinical trial tissue and blood samples from PARPi-treated SPOPmut and CHD1mut prostate cancer patients for biomarkers of this genetically driven phenotype (Aim 3).

项目概要/摘要 抑癌基因斑点型 POZ 蛋白 (SPOP)，cullin3-RING 泛素连接酶的底物接头， 表明高达 15% 的前列腺癌存在杂合错义突变，但其功能意义 这些 SPOP 突变的发生很大程度上是未知的。我们在多种蛋白质中发现了 SPOP 结合共有基序 规范的 cGAS-STING 和非规范的 STING-NF-κB 通路的研究，并证明人类和 小鼠 STING 蛋白是真正的 SPOP 靶点。 SPOP突变体（SPOPmut）和SPOP野生型分析 (SPOPwt) 去势抵抗性前列腺癌 (CRPC) 临床数据集揭示了 29 个基因“SPOPmut 基因” 签名”，反映了 STING-NF-κB 信号传导活性，并表明其在肿瘤招募中的作用 微环境 (TME) – 促进 SPOPmut CRPC 中肿瘤细胞的生长和存活。进一步评估 TCGA 中未经治疗的原发性前列腺癌的“SPOPmut 基因特征”揭示了 SPOP 突变体的一个子集 肿瘤，以及 CHD1 突变体 (CHD1mut) 肿瘤的子集，这些肿瘤的特征都得到了丰富。重要的是， SPOP 和 CHD1（染色质重塑因子）的共突变已有详细记录。稳定转导中 我们证明了人类和小鼠 SPOPmut（SPOPF102C 和 SPOPF133V）表达 CRPC 模型 非经典 STING-NF-κB-IL-6 通路蛋白、STAT3 和 HMG 蛋白的上调参与 促进分泌途径活动。我们还表明 PARP 抑制剂 (PARPi) 治疗 SPOPmut CRPC 细胞诱导 DNA 损伤，激活经典的 cGAS-STING-TBK-IFN-ß 信号传导并抑制非 典型的 STING-NF-κB-IL-6/STAT3，部分通过抑制性磷酸化 (S754-STAT3) 导致生长 抑制和凋亡信号传导。奥拉帕尼 (OLA) 增加 IFN-β 分泌并降低存活率 与巨噬细胞共培养与单一培养相比，SPOPmut 前列腺癌细胞中的程度高于对照细胞。 此外，仅在 OLA- 中证实了巨噬细胞中 cGAS-STING 的激活和 IFN-β 的诱导。 处理的共培养模型和中和抗体实验表明，OLA-的旁分泌调节 介导的生长抑制涉及 SPOPmut 前列腺癌细胞的 IFN-β 诱导和 IL-6 抑制。 我们假设前列腺癌中存在 SPOPmut、CHD1 缺失 (CHD1del) 和 SPOP + CHD1 共突变 通过增强合成致死率来促进 PARPi 或 PARPi + 抗 IL-6 治疗效果 未修复的 DNA 损伤增加，导致平衡向经典 cGAS-STING-IFN-ß 转变 IL-6/STAT3 的信号传导和抑制。我们建议分析 SPOPmut 的底层机制 和 CHD1del 前列腺癌表型，并定义这些基因驱动的 TME 改变及其蛋白质 使用最先进的蛋白质组学通过巨噬细胞重编程来产生效应子（目标 1）；分析功效 SPOPmut (F133V)、CHD1del 和 SPOPmut 中的 PARPi 和抗 IL-6 治疗；CHD1del RM-1-BM 同基因 CRPC 模型（目标 2）；并分析来自 PARPi 处理的 SPOPmut 的临床试验组织和血液样本， CHD1mut 前列腺癌患者的这种基因驱动表型的生物标志物（目标 3）。