Role of EPAC Signaling in Melanoma Progression

EPAC 信号传导在黑色素瘤进展中的作用

• 批准号: 9892600
• 负责人: Vijayasaradhi Setaluri
• 金额: --
• 依托单位: WM S. MIDDLETON MEMORIAL VETERANS HOSP
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-10-01 至 2023-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9892600
• 关键词: Adenylate Cyclase; Area; BRAF gene; Biochemical; CREB1 gene; Cell Cycle Progression; Cell Line; Cells; Cutaneous Melanoma; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Dependence; Development; Diagnosis; Disease; Disease-Free Survival; Distant; Dominant-Negative Mutation; Down-Regulation; Early Diagnosis; Early treatment; Excision; Exposure to; Fluorescence Resonance Energy Transfer; GTPase-Activating Proteins; Gene Expression; General Population; Generations; Genes; Goals; Growth; Growth and Development function; Guanine Nucleotide Exchange Factors; Homeostasis; Human; Immunotherapy; In Vitro; Incidence; Knowledge; Lead; Malignant Neoplasms; Manuscripts; Mediating; Melanoma Cell; Metabolic; Metabolism; Metastatic Melanoma; Military Personnel; Mitochondria; Molecular; Molecular Mechanisms of Action; Mus; Neoplasm Metastasis; Oncogenic; Organ; Oxidative Stress; PTEN gene; Pathway interactions; Patients; Pigments; Play; Primary Neoplasm; Production; Proteins; RAS Superfamily Proteins; Recurrence; Research; Risk; Risk Factors; Role; Sea; Services; Signal Pathway; Signal Transduction; Signaling Protein; Skin; Stress; Survival Rate; Testing; Tissue Microarray; UV Radiation Exposure; Ultraviolet Rays; Veterans; analog; base; cancer therapy; draining lymph node; experience; experimental study; in vivo; indexing; inhibitor/antagonist; knock-down; melanocyte; melanoma; mortality; mouse model; novel; overexpression; prevent; senescence; sensor; success; targeted treatment; transcription factor; transcriptome; transcriptome sequencing; tumor; tumor progression; tumorigenesis; ultraviolet

ABSTRACT Cutaneous melanoma arises from melanocytes in the skin. Exposure to solar ultraviolet (UV) radiation is a significant risk factor. Deployment of US military personnel around the world in areas of high UV index thus increases risk of melanoma during and after military service. If diagnosed early, excision of the primary tumor in the skin can be curative. However, the five-year survival rates decrease precipitously as melanoma metastasizes to local and regional lymph nodes and distant organs. Despite the recent successes and promise of targeted therapies and immunotherapy, early diagnosis and treatment of the localized primary cancer remain the best strategies to reduce cancer mortality. Therefore, a detailed understanding of the molecular mechanisms involved in melanoma tumor progression is critical for accurate and reliable diagnosis and treatment of aggressive cutaneous melanoma. Cyclic AMP (cAMP) signaling pathway plays a critical role in melanocyte proliferation and differentiation. This pathway is initiated upon elevation of intracellular cAMP by the catalytic activity of adenylyl cyclases and transduced through protein kinase A and transcription factor CREB culminating in expression of cAMP-responsive genes including the melanocyte master regulator, MITF. The role for cAMP signaling pathway in melanoma is not fully understood. Employing a human primary melanoma tissue microarray (TMA), a panel primary and metastatic melanoma cell lines and the BrafV600E/Pten-/- mouse model for melanoma, we recently demonstrated that cAMP signaling plays opposite roles in primary and metastatic melanoma cells, i. e., as a pro- proliferative signal in primary and as anti-proliferative signal in metastatic cells. We also showed that this switch in the cAMP of action is mediated by the alternative cAMP signaling axis involving EPAC-RAP1 (Exchange factor directly activated by cAMP- Ras-related protein 1). Additional studies showed that EPAC promotes cell cycle progression, activates mTORC1 signaling, and regulates cellular metabolism and mitochondrial ROS production selectively in primary melanoma cells. Furthermore, studies using a panel of primary and metastatic melanoma cell lines derived from the same patient revealed that the switch in the functions of EPAC occurs progressively during melanoma metastasis suggesting that EPAC is a critical component for the growth of primary melanoma and that rewiring of signaling networks during tumor progression abolish EPAC dependency in metastatic melanoma. Here, we propose to test the hypotheses that a) oncogenic transformation of melanocytes activates EPAC signaling and EPAC acts as a critical signaling node to suppress stress induced ROS generation, promote senescence escape and primary melanoma development, and b) gene expression changes and metabolic adaptions that occur during metastasis progressively abolish EPAC dependency. The specific aims of this application are: 1) Establish the role of EPAC in melanoma tumorigenesis and progression, 2) Determine the biochemical and molecular mechanisms of action of EPAC and 3) Investigate the role of EPAC in metabolic adaptation during melanoma progression. Successful completion of experiments proposed here could lead to identification of pathways and proteins that can be targeted to inhibit melanoma progression and treat metastatic melanoma. Therefore, we believe this project will have sustained impact in the field of melanoma.

抽象的 皮肤黑色素瘤来自皮肤中的黑色素细胞。暴露于太阳能紫外线（UV）辐射是 重要的风险因素。因此 在服兵役期间和之后增加黑色素瘤的风险。如果早点诊断，请切除原发性肿瘤 皮肤可以治愈。但是，五年生存率随着黑色素瘤转移而急剧降低 到局部和区域淋巴结和遥远的器官。尽管最近取得了成功和有望 疗法和免疫疗法，局部原发性癌症的早期诊断和治疗仍然是最好的 降低癌症死亡率的策略。因此，对所涉及的分子机制的详细理解 在黑色素瘤中，肿瘤进展对于准确可靠的诊断和侵略性治疗至关重要 皮肤黑色素瘤。环状AMP（CAMP）信号通路在黑素细胞增殖中起着至关重要的作用 分化。该途径是通过腺细胞的催化活性在升高细胞内cAMP的情况下开始的 循环酶并通过蛋白激酶A和转录因子CREB ​​CRMIND在表达中的转导酶 营地响应基因，包括黑素细胞主调节器Mitf。营地信号通路的作用 在黑色素瘤中尚不完全了解。采用人类原发性黑色素瘤微阵列（TMA），一个面板 原发性和转移性黑色素瘤细胞系以及黑色素瘤的BRAFV600E/PTEN - / - 小鼠模型，我们最近 证明cAMP信号在原发性和转移性黑色素瘤细胞中起相反的作用，即。 e。，作为亲人 在转移细胞中的原发性和抗增殖信号中的增生信号。我们还表明了这个开关 在动作营地中，涉及epac-rap1（交换系数）的替代cAMP信号轴介导 直接被与营地相关蛋白1）直接激活。其他研究表明，EPAC促进了细胞周期 进展，激活MTORC1信号并调节细胞代谢和线粒体ROS的产生 在原发性黑色素瘤细胞中有选择地。此外，使用一组原发性和转移性黑色素瘤的研究 来自同一患者的细胞系显示EPAC功能的转换逐渐发生 在黑色素瘤转移期间，表明EPAC是原发性黑色素瘤生长的关键成分 肿瘤进展过程中信号网络的重新布线取消了转移性的EPAC依赖性 黑色素瘤。在这里，我们建议测试a）黑素细胞致癌转化的假设。 EPAC信号传导和EPAC充当抑制应力诱导ROS产生的关键信号节点，促进 衰老逃逸和原发性黑色素瘤的发育，b）基因表达变化和代谢 转移期间发生的适应性逐渐消除了EPAC的依赖性。这个特定的目的 应用是：1）确定EPAC在黑色素瘤肿瘤发生和进展中的作用，2）确定 EPAC的作用和3）研究EPAC在代谢中的作用 黑色素瘤进展过程中的适应性。成功完成此处提出的实验可能会导致 可以鉴定可针对抑制黑色素瘤进展并治疗转移性的途径和蛋白质 黑色素瘤。因此，我们认为该项目将在黑色素瘤领域持续影响。