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).

项目总结/摘要 肿瘤抑制基因斑点型POZ蛋白（SPOP），cullin 3-RING泛素连接酶的底物衔接子， 在高达15%的前列腺癌中显示杂合错义突变，但功能意义 这些SPOP突变在很大程度上未知。我们在多种蛋白质中鉴定了SPOP结合共有基序 经典cGAS-STING和非经典STING-NF-κB通路的表达，并证明人和 小鼠STING蛋白是真正的SPOP靶标。SPOP突变体（SPOPmut）和SPOP野生型的分析 （SPOPwt）去势抵抗性前列腺癌（CRPC）的临床数据集显示，29个基因的“SPOPmut基因 STING-NF-κB信号通路的活性，提示STING-NF-κB信号通路在肿瘤细胞募集中的作用 在SPOPmut CRPC中，微环境（TME）促进肿瘤细胞生长和存活。进一步评估 来自TCGA的原发性未经治疗的前列腺癌中的“SPOPmut基因签名”揭示了SPOP突变体的一个子集 肿瘤，以及CHD 1突变体（CHD 1 mut）肿瘤的子集，其富集了该特征。重要的是， SPOP和CHD 1（染色质重塑因子）中的共突变被充分记载。在稳定转导的 人和小鼠SPOPmut（SPOPF 102 C和SPOPF 133 V）表达CRPC模型，我们证明了 非经典STING-NF-κB-IL-6通路蛋白、STAT 3和HMG蛋白的上调参与了 促进分泌途径活性。我们还表明，PARP抑制剂（PARPi）治疗SPOPmut CRPC细胞诱导DNA损伤，激活典型的cGAS-STING-TBK-IFN-γ信号传导并抑制非细胞毒性。 典型的STING-NF-κB-IL-6/STAT 3，部分通过抑制磷酸化（S754-STAT 3），导致生长 抑制和凋亡信号传导。奥拉帕尼（奥拉）增加IFN-β分泌并降低存活率， SPOPmut前列腺癌细胞与巨噬细胞共培养与单培养相比， 此外，仅在奥拉中证实巨噬细胞中cGAS-STING的活化和IFN-γ的诱导。 处理的共培养模型，中和抗体实验表明，旁分泌调节奥拉- 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SPOPmut、CHD 1缺失（CHD 1del）和SPOP + CHD 1共突变 通过增强的合成致死性促进PARPi或PARPi +抗IL-6治疗功效， 增加的、未修复的DNA损伤，导致平衡向典型的cGAS-STING-IFN-γ转移 信号传导和抑制IL-6/STAT 3。我们建议分析SPOPmut的潜在机制 和CHD 1del前列腺癌表型，并定义这些遗传驱动的TME改变及其蛋白 使用最先进的蛋白质组学通过巨噬细胞重编程效应器（目的1）;分析 SPOPmut（F133 V）、CHD 1del和SPOPmut中的PARPi和抗IL-6治疗; CHD 1del RM-1-BM同基因 CRPC模型（目的2）;并分析来自PARPi治疗的SPOPmut和SPOPmut的临床试验组织和血液样品。 CHD 1 mut前列腺癌患者的这种遗传驱动表型的生物标志物（目的3）。