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受体相关激酶-M（IRAK-1）与IL-1受体相关激酶-M（IL-1 receptor associated kinase-M，IRAK-1）的相互作用机制。 M）在黑色素瘤中的表达调节转移，并开发临床相关的方法来诱导和 通过恢复IRAK-M表达维持抗肿瘤T细胞免疫。癌症转移是导致 癌症的死亡率和发病率，占癌症相关死亡的90%。免疫治疗已 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。 患者然而，尽管取得了这些成功，临床受益仅限于一小部分患者。一个 免疫治疗的潜在和普遍障碍是多种免疫抑制机制， 在很大程度上源于炎症信号的慢性表达。理解基本的 调节这些因子并靶向这些途径的机制对于开发有效的 预防或减少转移和恢复抗肿瘤T细胞活性的策略。我们之前报道过， IL-1受体相关激酶-4（IRAK-4）在黑素瘤中过表达和活化。IRAK-4是一种 炎症过程中的中枢激酶，也调节促炎性和 免疫抑制分子（包括ATP、VEGF、PDGF、IL-1）。这一信号通路被IL- 1受体和Toll样受体。我们之前报道过抑制黑色素瘤中的IRAK-4活性 显著降低许多这些炎症因子的表达， 小鼠通过检测通常抑制IRAK-4信号传导的内源性蛋白质的水平，我们发现IRAK-M 在黑色素瘤中缺乏（患者和细胞系）。IRAK-M在IRAK家族成员中独一无二， 它是炎性IRAK-4信号传导的负调节剂。我们发现，恢复IRAK-M的表达， 黑色素瘤细胞显著降低了它们的侵袭（体外）和转移（临床前模型）能力。 这与细胞运动性相关的关键分子表达水平的变化有关， 细胞骨架动力学和非癌细胞中板状伪足的形成，但它们在黑色素瘤或 转移未知。这些分子包括无远端同源异型盒5（DLX 5;一种转录因子， 激活和抑制活性）、整联蛋白ITGA 11和细胞质FMR 1相互作用蛋白2（Cyfip 2）。 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，同时抑制了T细胞功能障碍。 IRAK-4增强抗肿瘤T细胞活性。IRAK-4改变T细胞活性的能力与其抑制T细胞增殖的能力有关。 调节细胞外ATP水平; ATP的代谢物腺苷强烈抑制T细胞活性。通过 在这些研究中，我们将检验黑色素瘤中恢复IRAK-M表达会损害其细胞增殖的假设。 通过调节DLX 5的表达，DLX 5反过来调节细胞侵袭，从而提高转移潜力。我们 进一步假设癌细胞中IRAK-M的表达通过降低肿瘤细胞的增殖来恢复抗肿瘤T细胞活性， 细胞外ATP的表达水平和信号来源。在目标1中，我们将确定 IRAK-M调节癌细胞侵袭的机制。这一目标建立在令人信服的初步数据基础上 这表明癌细胞中的IRAK-M诱导损害了它们通过基质或内皮细胞侵入的能力， 细胞（体外）和转移（体内）。我们推测DLX 5上调ITGA 11，但下调 Cyfip 2表达从而控制细胞迁移、细胞形态发生和侵袭。通过目标2，我们 提出通过恢复肿瘤细胞中的IRAK-M信号传导来恢复肿瘤免疫。我们将使用基因 方法和预定义的药物组，选择性诱导黑色素瘤中的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 提供关于IRAK-M如何调节癌细胞转移的机制见解的潜力， 开发临床相关的新策略以恢复肿瘤免疫的机会。