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: 10292971
• 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/10292971
• 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; Signal Pathway; Signal Transduction; Signaling Protein; Skin; Stress; Survival Rate; Testing; Tissue Microarray; UV Radiation Exposure; Veterans; active duty; analog; base; cancer therapy; draining lymph node; experience; experimental study; in vivo; indexing; inhibitor; knock-down; melanocyte; melanoma; military service; military veteran; mortality; mouse model; novel; overexpression; prevent; senescence; sensor; solar ultraviolet radiation; 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 皮肤黑色素瘤源自皮肤中的黑色素细胞。暴露于太阳紫外线 (UV) 辐射是一种 显着的风险因素。美军在全球高紫外线指数地区部署军事人员 服兵役期间和服兵役后增加患黑色素瘤的风险。如果早期诊断，可切除原发肿瘤 皮肤可以有疗效。然而，随着黑色素瘤的转移，五年生存率急剧下降 到局部和区域淋巴结和远处器官。尽管最近取得了成功并承诺有针对性 疗法和免疫疗法，局部原发癌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，详细了解所涉及的分子机制 在黑色素瘤中，肿瘤进展对于侵袭性黑色素瘤的准确、可靠的诊断和治疗至关重要 皮肤黑色素瘤。环磷酸腺苷 (cAMP) 信号通路在黑色素细胞增殖和 差异化。该途径是在腺苷酸催化活性导致细胞内 cAMP 升高时启动的。 环化酶并通过蛋白激酶 A 和转录因子 CREB ​​转导，最终表达 cAMP 反应基因，包括黑素细胞主调节因子 MITF。 cAMP信号通路的作用 在黑色素瘤中的作用尚不完全清楚。采用人类原发性黑色素瘤组织微阵列 (TMA)，一组 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发挥相反的作用，即。即，作为亲 原代细胞中的增殖信号和转移细胞中的抗增殖信号。我们还表明这个开关 cAMP 的作用是由涉及 EPAC-RAP1（交换因子）的替代 cAMP 信号轴介导的 由 cAMP-Ras 相关蛋白直接激活 1)。其他研究表明 EPAC 促进细胞周期 进展，激活 mTORC1 信号传导，并调节细胞代谢和线粒体 ROS 产生 选择性地作用于原代黑色素瘤细胞。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 功能的转变是逐渐发生的 在黑色素瘤转移过程中，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消除了转移性肿瘤中的 EPAC 依赖性 黑色素瘤。在这里，我们建议测试以下假设：a) 黑素细胞的致癌转化激活 EPAC信号传导和EPAC作为关键的信号传导节点来抑制应激诱导的ROS生成，促进 衰老逃逸和原发性黑色素瘤发展，b) 基因表达变化和代谢 转移过程中发生的适应逐渐消除 EPAC 依赖性。本次活动的具体目标 应用包括：1) 确定 EPAC 在黑色素瘤发生和进展中的作用，2) 确定 EPAC 的生化和分子作用机制以及 3) 研究 EPAC 在代谢中的作用 黑色素瘤进展期间的适应。成功完成此处提出的实验可能会导致 鉴定可靶向抑制黑色素瘤进展和治疗转移性的途径和蛋白质 黑色素瘤。因此，我们相信该项目将对黑色素瘤领域产生持续影响。