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).

项目摘要/摘要 肿瘤抑制基因斑点型POZ蛋白（SPOP），Cullin3环泛素连接酶的底物适配器， 在多达15％的前列腺癌中证明了杂合的错义突变，但功能意义 这些汤匙突变在很大程度上是未知的。我们鉴定了多种蛋白质中的Spop结合共识基序 规范的CGAS丁字和非典型的sting-nf-κB途径，并证明了人类和 小鼠刺激蛋白是真正的SPOP靶标。 SPOP突变体（Spopmut）和Spop野生型的分析 （SPOPWT）耐castration-抗性前列腺癌（CRPC）临床数据集揭示了29基因“ Spopmut基因 签名”，反映了STING-NF-κB信号传导活性，并提出了肿瘤募集的作用 微环境（TME） - Spopmut CRPC中促进肿瘤细胞的生长和存活。进一步评估 来自TCGA的原发性，未处理的前列腺癌中的“ Spopmut基因签名”显示了一部分Spop突变体 肿瘤以及CHD1突变体（CHD1MUT）肿瘤的一部分，这些肿瘤被富含用于特征的肿瘤。重要的是， 索波和CHD1（染色质重塑因子）中的共突变已充分记录。在稳定的翻译中 人和小鼠spopmut（Spopf102c和spopf133v） - 表达CRPC模型，我们证明了 与参与参与 促进秘密途径活动。我们还表明了pAPMUT的PARP抑制剂（PARPI）治疗 CRPC细胞诱导DNA损伤，激活规范的CGAS-sting-TBK-IFN-ß信号，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。 Olaparib（OLA）增加了IFN-β分泌并降低了生存能力 spopmut前列腺癌细胞的范围比巨噬细胞与单一培养的共培养细胞中的对照细胞中的程度。 另外，仅在OLA-中证明了CGAS刺的激活和IFN-ß的诱导。 经过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IFN-β诱导和spopmut前列腺癌细胞的IL-6抑制。 我们假设Popmut，CHD1缺失（CHD1DEL）和SPOP + CHD1在前列腺中的共透性取消 通过增强的合成致死率通过 增加，未修复的DNA损伤，导致平衡向规范CGAS-sting-IFN-ß转移 IL-6/STAT3的信号传导和抑制。我们建议分析spopmut的基本机制 和CHD1DEL前列腺癌表型，并定义这些一般驱动的TME改变及其蛋白 使用最先进的蛋白质组学重新编程通过巨噬细胞重编程（AIM 1）；分析有效性 Spopmut（F133V），CHD1DEL和SPOPMUT中的PARPI和抗IL-6处理； CRPC模型（AIM 2）；并分析来自parpi处理的spopmut的临床试验组织和血液样本 CHD1MUT前列腺癌患者用于这种一般驱动的表型的生物标志物（AIM 3）。