Restoring the Regulation of a Central Signaling Pathway to Prevent Metastases and Reinstate Tumor Immunity

恢复中央信号通路的调节以预防转移并恢复肿瘤免疫

• 批准号: 10454780
• 负责人: Eduardo V Davila
• 金额: --
• 依托单位: VA EASTERN COLORADO HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10454780
• 关键词: ADORA2A gene; Accounting; Adenosine; Age; Cancer Etiology; Cell Line; Cell Mobility; Cells; Cellular Morphology; Cessation of life; Chronic; Clinical; Cytoskeleton; Data; Development; Disseminated Malignant Neoplasm; Distal; Endothelial Cells; Engineering; Exhibits; FDA approved; FMR1; Family; Family member; Functional disorder; Gene Chips; Generations; Genes; Genetic; Genetic Transcription; Goals; Homeobox; Human; IRAK1 gene; ITGA11 gene; Immunity; Immunotherapy; Impairment; In Vitro; Inflammatory; Integrins; Interleukin-1; Interleukin-1 Receptors; Interleukin-1 beta; Interruption; Invaded; Lead; Link; Malignant Neoplasms; Mediating; Medical; Melanoma Cell; Metastatic Melanoma; Metastatic to; Modeling; Molecular; Morbidity - disease rate; Morphogenesis; Mus; Myeloid-derived suppressor cells; Names; Neoplasm Metastasis; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphotransferases; Platelet-Derived Growth Factor; Play; Population; Pre-Clinical Model; Process; Production; Property; Proteins; Recurrence; Regulation; Reporting; Resistance; Role; Sampling; Signal Pathway; Signal Transduction; Survival Rate; T cell response; T cell therapy; T-Lymphocyte; Testing; Therapeutic; Toll-like receptors; Tumor Immunity; Vascular Endothelial Growth Factors; Veterans; anti-tumor immune response; base; cancer cell; cancer immunotherapy; cancer therapy; cancer type; cell motility; clinically relevant; effective therapy; exhaustion; experience; extracellular; genetic approach; high risk; in vivo; insight; interleukin-1 receptor-associated kinase; knock-down; melanoma; member; metastatic process; mortality; neoplastic cell; novel; novel strategies; overexpression; prevent; receptor; standard of care; success; treatment strategy; tumor; tumor growth; tumor microenvironment; tumor progression

Our long-term goals are to understand the mechanisms by which IL-1 receptor associated kinase-M (IRAK- M) expression in melanoma regulates metastasis and to develop clinically-relevant approaches to induce and sustain anti-tumor T cell immunity by restoring IRAK-M expression. Cancer metastasis is the main cause of cancer mortality and morbidity, accounting for 90% of cancer-related deaths. Immunotherapy has revolutionized the treatment of advanced (metastatic) melanoma and is now the standard of care for most patients. Despite these successes however, clinical benefits are restricted to small percentage of patients. An underlying and universal hindrance to immunotherapies is a variety of immunosuppressive mechanisms that in large part originate from the chronic expression of inflammatory signals. Understanding the fundamental mechanisms that regulate these factors and targeting these pathways is critical for developing effective strategies to prevent or reduce metastases and to restore antitumor T cell activity. We previously reported that the IL-1 receptor associated kinase-4 (IRAK-4) is overexpressed and activated in melanoma. IRAK-4 is a central kinase in the inflammatory process that also regulates the expression of metastases-promoting and immunosuppressive molecules (including ATP, VEGF PDGF, IL-1). This signaling pathway is activated by IL- 1 receptor and toll-like receptors. We previously reported that inhibiting IRAK-4 activity in melanoma drastically reduces the expression of many of these inflammatory factors and reduces cancer progression in mice. By examining the levels of endogenous proteins that normally inhibit IRAK-4 signaling we found IRAK-M to be deficient in melanomas (patients and cell lines). IRAK-M is unique among the IRAK family members in that it is a negative regulator of inflammatory IRAK-4 signaling. We found that restoring IRAK-M expression in melanoma cells drastically reduced their ability to invade (in vitro) and to metastasize (in preclinical models). This was associated with changes in the expression levels of key molecules associated with cell mobility, cytoskeletal dynamics, and formation of lamellipodium in non-cancerous cells but their role in melanoma or metastasis is unknown. These molecules include Distal-Less Homeobox 5 (DLX5; a factor with transcription activating and repressing activity), the integrin ITGA11, and cytoplasmic FMR1 Interacting Protein 2 (Cyfip2). Furthermore, we observed that IRAK-4 signaling in melanoma accelerated T cell dysfunction while inhibiting IRAK-4 enhanced antitumor T cell activity. IRAK-4’s ability to alter T cell activity was associated with its ability to regulate extracellular ATP levels; ATP’s metabolite, adenosine, strongly suppresses T cell activity. Through these studies, we will test the hypothesis that restoring IRAK-M expression in melanoma impairs their metastatic potential by regulating the expression of DLX5 which in turn regulates cell invasion. We further postulate that IRAK-M expression in cancer cells restores antitumor T cell activity by reducing the expression levels of and signals originating from extracellular ATP. In Aim 1, we will determine the mechanism by which IRAK-M regulates cancer cell invasion. This aim builds on compelling preliminary data indicating that IRAK-M induction in cancer cells impairs their ability to invade through matrix or endothelial cells (in vitro) and to metastasize (in vivo). We conjecture that DLX5 upregulates ITGA11 but downregulates Cyfip2 expression thereby controlling cell migration, cell morphogenesis and invasion. Through Aim 2, we propose to restore tumor immunity by reinstating IRAK-M signaling in tumor cells. We will use genetic approaches and a predefined panel of drugs, that selectively induce IRAK-M in melanoma, to investigate the cellular and molecular mechanisms by which IRAK-M enhances anti-tumor T cell activity. These studies have the potential to provide mechanistic insights as to how IRAK-M regulate cancer cell metastasis and opportunities to develop clinically relevant, novel strategies to restore tumor immunity.

我们的长期目标是了解IL-1接收器与激酶-M相关的机制（irak-- M）在黑色素瘤中的表达调节转移，并开发出与临床相关的诱导和 通过恢复IRAK-M表达来维持抗肿瘤T细胞免疫。癌症转移是 癌症死亡率和发病率，占癌症相关死亡的90％。免疫疗法具有 彻底改变了晚期（转移性）黑色素瘤的治疗，现在是大多数人的护理标准 患者。尽管取得了这些成功，但临床益处仅限于少数患者。一个 免疫疗法的基本和普遍障碍是多种免疫抑制机制 在很大程度上，源自炎症信号的慢性表达。了解基本 调节这些因素并瞄准这些途径的机制对于发展有效的机制至关重要 预防或减少转移并恢复抗肿瘤T细胞活性的策略。我们以前报道了 IL-1受体相关的激酶-4（IRAK-4）在黑色素瘤中过表达并激活。 irak-4是一个 中央激酶在炎症过程中也调节转移酶的表达和 免疫抑制分子（包括ATP，VEGF PDGF，IL-1）。该信号通路被IL-激活 1个受体和收费受体。我们先前报道说，抑制黑色素瘤的IRAK-4活性 大幅度降低了许多这些炎症因子的表达，并降低了癌症的进展 老鼠。通过检查通常抑制IRAK-4信号的内源性蛋白水平，我们发现了Irak-M 缺乏黑色素瘤（患者和细胞系）。 Irak-M在Irak家庭成员中是独一无二的 它是炎症性IRAK-4信号传导的负调节剂。我们发现恢复Irak-M的表达 黑色素瘤细胞大大降低了其入侵（体外）和转移的能力（在临床前模型中）。 这与与细胞迁移率相关的关键分子的表达水平的变化有关， 细胞骨架动力学和非癌细胞中层状植物的形成，但它们在黑色素瘤或 转移是未知的。这些分子包括远端的同源蛋白蛋白蛋白蛋白酶5（DLX5；带有转录的因素 激活和反映活性），整联蛋白ITGA11和细胞质FMR1相互作用蛋白2（CYFIP2）。 此外，我们观察到黑色素瘤中的IRAK-4信号传导加速了T细胞功能障碍，同时抑制 IRAK-4增强的抗肿瘤T细胞活性。 IRAK-4改变T细胞活动的能力与其能力有关 调节细胞外ATP水平； ATP的代谢产物腺苷强烈抑制T细胞活性。通过 这些研究，我们将测试以下假设：恢复黑色素瘤中的IRAK-M表达会损害其 通过调节DLX5的表达来调节细胞侵袭，转移潜力。我们 进一步假设癌细胞中的IRAK-M表达通过减少 来自细胞外ATP的信号的表达水平和信号。在AIM 1中，我们将确定 IRAK-M通过调节癌细胞侵袭的机制。此目标以引人注目的初步数据为基础 表明癌细胞中的IRAK-M诱导会损害其通过基质或内皮侵袭的能力 细胞（体外）并转移（体内）。我们猜想DLX5上调了ITGA11，但下调了 CYFIP2表达，从而控制细胞迁移，细胞形态发生和侵袭。通过AIM 2，我们 通过恢复肿瘤细胞中的IRAK-M信号传导来恢复肿瘤免疫的建议。我们将使用遗传 接近和预定义的药物，有选择地影响黑色素瘤的Irak-M，以研究 IRAK-M增强抗肿瘤T细胞活性的细胞和分子机制。这些研究有 关于IRAK-M如何调节癌细胞转移和 开发临床相关的新型策略以恢复肿瘤免疫学的机会。