The integrated stress response and the microenvironment in melanoma progression

黑色素瘤进展中的综合应激反应和微环境

• 批准号: 10391719
• 负责人: Colin Goding
• 金额: $ 43.03万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10391719
• 关键词: 3-Dimensional; Ablation; Adipocytes; Allografting; Anoikis; Antioxidants; Apoptosis; Apoptotic; Autophagocytosis; BRAF gene; Binding; Blood Vessels; CAV1 gene; Cells; Coculture Techniques; Collagen; Complex; DNA Sequence Alteration; Endothelial Cells; Endothelium; Equilibrium; Experimental Neoplasms; Exposure to; Extracellular Matrix; Fatty Acid Desaturases; Fatty Acids; Fibroblasts; Gene Expression; Genes; Genetic; Genetic Transcription; Growth; Heterogeneity; Human; Immunotherapy; Impairment; Inflammation; Integrins; Knockout Mice; Lead; Link; Lymph; Mediating; Mediator of activation protein; Melanoma Cell; Metastatic Neoplasm to the Lung; Modality; Modeling; Monounsaturated Fatty Acids; Mus; Neoplasm Metastasis; Non-Malignant; Nuclear; Oncogenes; Oncogenic; Output; Oxygen; Pathway interactions; Patients; Persons; Pharmacology; Phenotype; Primary Neoplasm; Publishing; Recovery; Refractory; Reporting; Resistance; Role; Shapes; Signal Pathway; Signal Transduction; Stress; Testing; Therapeutic; Therapeutic Intervention; Tumor Suppressor Proteins; Tumor-infiltrating immune cells; Xenograft procedure; angiogenesis; biological adaptation to stress; endoplasmic reticulum stress; gain of function; immune checkpoint blockade; improved; inhibitor; melanoma; microphthalmia-associated transcription factor; mouse model; neoplastic cell; new therapeutic target; novel; novel therapeutics; nutrient deprivation; programs; response; targeted agent; targeted treatment; transcription factor; tumor; tumor growth; tumor microenvironment; tumorigenic; uptake

SUMMARY Despite the emergence novel therapeutic modalities, including BRAF inhibitors and immunotherapies, over 7,100 people are expected to die each year from malignant melanoma, primarily from metastatic dissemination and therapy resistance. The proposed studies leverage the expertise of the two co-PIs in tumor microenvironment, the Integrated Stress Response (ISR) and melanoma progression to test the overall hypothesis that as part of the adaptive response to tumor microenvironmental (TME) stress, bi-directional interactions between melanoma cells and fibroblasts, endothelial cells and adipocytes promote survival, metastatic dissemination, and therapy resistance. Preliminary results and published reports from the two PIs indicate that the ISR is activated in human melanomas and that genetic or pharmacological disruption of the ISR severely impairs primary tumor growth and metastasis in multiple experimental tumors, including melanomas. The studies will delineate the salient roles of the transcription factor ATF4, a major transcriptional effector of the Integrated Stress Response, in a pro-survival and pro-metastatic program mediated by the non- canonical tumor suppressor BRN2, a transcription factor effector of multiple melanoma-associated signaling pathways. To test the central hypothesis, we will pursue three specific aims: In Aim 1 we will use a novel, conditional global ATF4 knockout mouse model, as well as Fibroblast (FB)-specific and Endothelial (EC)- specific ATF4 knockout mice and cells to determine the role of ATF4 expression in each TME component on melanoma progression and metastasis. Preliminary results with global or FB-specific deletion of ATF4 results in a severe deficiency in tumor growth of flank melanoma tumors. In Aim 2 we will study how SRC promotes melanoma progression via ATF4-BRN2 cooperativity. Both BRN2 and ATF4 can repress anoikis/apoptosis, and in preliminary studies we reveal that BRN2 interacts with ATF4. Therefore, we will test the hypothesis that Extracellular Matrix (ECM)-driven integrin signaling, and monounsaturated fatty acid (MUFA) uptake from adipocytes and lymph converge on SRC to impose a cooperative BRN2-ATF4 anti-apoptotic and pro- metastasis gene expression program, driven in part by hippo signaling. Finally, under Aim 3, we will determine how MUFAs dictate melanoma phenotype. Here we will dissect a novel mechanism underpinning MUFA- mediated nuclear localization of -catenin and test the hypothesis that the resulting SRC-driven nuclear CAV1- -catenin complex promotes a pro-metastasis gene expression program via BRN2-ATF4, and the contribution of ATF4 and BRN2 to melanoma phenotypic heterogeneity and tumor immune infiltration. By delineating how the ISR uses ATF4 to coordinate the output of the TME to shape melanoma progression, we will identify therapeutically exploitable pathways for anti-melanoma approaches.

总结 尽管出现了新的治疗方式，包括BRAF抑制剂和免疫疗法， 预计每年有7,100人死于恶性黑色素瘤，主要是转移性传播 和治疗抵抗。拟议的研究利用了两个共同PI在肿瘤领域的专业知识 微环境，综合应激反应（ISR）和黑色素瘤进展，以测试整体 假设作为对肿瘤微环境（TME）应激的适应性反应的一部分， 黑素瘤细胞与成纤维细胞、内皮细胞和脂肪细胞之间的相互作用促进存活， 转移性扩散和治疗抗性。两名PI的初步结果和发表的报告 表明ISR在人黑色素瘤中被激活， ISR严重损害多种实验性肿瘤的原发性肿瘤生长和转移，包括 黑素瘤。这些研究将描述转录因子ATF 4的突出作用，ATF 4是一种主要的转录因子， 在由非应激因子介导的促生存和促转移程序中，综合应激反应的效应子 典型的肿瘤抑制因子BRN 2，一种多个黑色素瘤相关信号传导的转录因子效应子 途径。为了检验中心假设，我们将追求三个具体目标：在目标1中，我们将使用一部小说， 条件性整体ATF 4敲除小鼠模型，以及成纤维细胞（FB）特异性和内皮细胞（EC）特异性 特定的ATF 4敲除小鼠和细胞，以确定ATF 4表达在每个TME组分中的作用， 黑素瘤进展和转移。全局或FB特异性缺失ATF 4结果的初步结果 侧腹黑色素瘤肿瘤生长严重不足。在目标2中，我们将研究SRC如何促进 通过ATF 4-BRN 2协同作用的黑素瘤进展。BRN 2和ATF 4都可以抑制失巢凋亡/凋亡， 在初步研究中，我们发现BRN 2与ATF 4相互作用。因此，我们将检验以下假设： 细胞外基质（ECM）驱动的整合素信号传导和来自细胞外基质的单不饱和脂肪酸（MUFA）摄取 脂肪细胞和淋巴细胞聚集在SRC上，以施加合作的BRN 2-ATF 4抗凋亡和促凋亡作用。 转移基因表达程序，部分由hippo信号驱动。最后，根据目标3，我们将确定 MUFA如何决定黑色素瘤表型。在这里，我们将剖析一个新的机制，支持MUFA- 介导的β-连环蛋白的核定位，并测试假设，所产生的SRC驱动的核CAV 1- β-连环蛋白复合物通过BRN 2-ATF 4促进促转移基因表达程序， ATF 4和BRN 2与黑色素瘤表型异质性和肿瘤免疫浸润的关系。通过描述如何 ISR使用ATF 4协调TME的输出，以塑造黑色素瘤进展，我们将识别 抗黑色素瘤方法的治疗上可利用的途径。