Project 1 - Siah 1 in ER and Mitochondrial Function and Homeostasis in Melanoma

项目 1 - Siah 1 在黑色素瘤中的 ER 和线粒体功能及稳态

• 批准号: 9925090
• 负责人: Ze'ev A Ronai
• 金额: $ 39.68万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2021-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9925090
• 关键词: Animals; Apoptotic; Autophagocytosis; BRAF gene; Biogenesis; Bioinformatics; Cell Aging; Clinical; Congenic Mice; Data; Drug resistance; Elements; Endoplasmic Reticulum; Engineering; Exhibits; Foundations; Genes; Genetic; Genetic Models; Genetic Transcription; Homeostasis; Lead; Link; MEKs; Melanoma Cell; Metabolic; Metabolic Pathway; Mitochondria; Modeling; Neoplasm Metastasis; Outcome; Oxidative Stress; Pathway interactions; Patients; Pharmacotherapy; Phenotype; Play; Protein Isoforms; Proteins; Regulation; Resistance; Role; Signal Transduction; Stimulus; Stress; Testing; Up-Regulation; Work; base; chemotherapy; clinically significant; cohort; congenic; endoplasmic reticulum stress; inhibitor/antagonist; insight; knock-down; melanocyte; melanoma; mitochondrial dysfunction; mutant; neoplastic cell; programs; protein folding; protein misfolding; response; sensor; therapy resistant; treatment response; tumor; tumor heterogeneity; ubiquitin ligase

SUMMARY – PROJECT 1: SIAH1 IN ER AND MITOCHONDRIAL FUNCTION AND HOMEOSTASIS IN MELANOMA   Growing evidence indicates that activation of the adaptive unfolded protein response (UPR) occurs in melanoma cells following drug therapy. The UPR senses oxidative stress, mitochondrial dysfunction and dynamics and resolve protein misfolding in the ER, activities often deregulated in melanoma tumor cells. Notably, the degree of UPR signaling differs among melanoma subtypes, which is consistent with tumor heterogeneity and plasticity and underlies the propensity to adapt to chemotherapy and related stress stimuli. More recently, tumor resistance has been associated with altered expression and activity of the master regulators PGC1α and MITF, factors central for mitochondrial and melanocyte/melanoma biogenesis, respectively. Recent studies, including our preliminary results, demonstrate that deregulation of effectors of the UPR, including ATF4, CHOP and IRE1α, plays a fundamental role in melanoma. Work supported by this P01 established that the ubiquitin ligases Siah1/2 are critical to amplify UPR signaling and identified Siah1 isoform 2 (Siah1is2) as the major isoform induced by the BRAFi (PLX4032) and correlated with PGC1α and MITF expression. Our emerging model is that Siah1is2 is a key sensor that finely tunes the ATF4–IRE1α regulatory axis to control PGC1α and MITF activities, key elements in the propensity of melanoma to adapt to environmental conditions associated with drug resistance and metastatic capacity. Efforts coordinated with Projects 2 and 3 and Cores B and C will further define regulation of PGC1α and MITF expression and activity and evaluate the contribution of ATF4, CHOP, and Siah1is2 to therapeutic responses. To test the hypothesis that UPR- and Siah1is2-controlled signaling engages and fine-tunes the PGC1α and MITF regulatory network to alter metabolic and transcriptional programs, which underlie melanoma resistance and metastasis. We proposed to determine (i) how does Siah1is2 control of the ATF4–PGC1α–MITF regulatory axis define melanoma resistance and metastatic phenotypes and (ii) how does ATF4/CHOP regulate melanoma metastasis and drug resistance? Our studies will rely on congenic melanoma tumors, naive or resistant to therapy, congenic Braf/Pten derived melanoma lines CRSPER'd for UPR and Siah1is2 genes, and genetic melanoma models crossed with Atf4, Chop or Siah1 mutant animals. Our studies are expected to lead to unprecedented new insight into the precise role select UPR components play in control of melanoma plasticity, underlying its propensity to resistance and metastatic phenotypes—which represent a key unmet clinical need.

摘要 – 项目 1：ER 中的 SIAH1 和线粒体功能以及体内平衡 黑色素瘤   越来越多的证据表明，适应性未折叠蛋白反应（UPR）的激活发生在 药物治疗后的黑色素瘤细胞。 UPR 感知氧化应激、线粒体功能障碍和 动力学并解决内质网中蛋白质错误折叠的问题，这些活性在黑色素瘤肿瘤细胞中经常失调。 值得注意的是，UPR信号传导的程度在黑色素瘤亚型之间存在差异，这与肿瘤的发生一致。 异质性和可塑性是适应化疗和相关应激刺激的倾向的基础。 最近，肿瘤耐药性与主基因表达和活性的改变有关。 调节剂 PGC1α 和 MITF，线粒体和黑色素细胞/黑色素瘤生物发生的核心因子， 分别。最近的研究，包括我们的初步结果，表明，对效应器的放松管制 UPR，包括 ATF4、CHOP 和 IRE1α，在黑色素瘤中发挥着重要作用。本 P01 支持的工作 确定泛素连接酶 Siah1/2 对于放大 UPR 信号传导至关重要，并鉴定了 Siah1 亚型 2 (Siah1is2) 作为 BRAFi (PLX4032) 诱导的主要亚型并与 PGC1α 和 MITF 相关 表达。我们的新兴模型是 Siah1is2 是一个关键传感器，可以微调 ATF4–IRE1α 监管 控制 PGC1α 和 MITF 活性的轴，这是黑色素瘤适应倾向的关键要素 环境条件与耐药性和转移能力相关。协调努力 项目 2 和 3 以及核心 B 和 C 将进一步定义 PGC1α 和 MITF 表达和活性的调节 并评估 ATF4、CHOP 和 Siah1is2 对治疗反应的贡献。检验假设 UPR 和 Siah1is2 控制的信号传导参与并微调 PGC1α 和 MITF 调节 网络改变代谢和转录程序，这是黑色素瘤抵抗和 转移。我们建议确定 (i) Siah1is2 如何控制 ATF4–PGC1α–MITF 调节 轴定义黑色素瘤耐药性和转移表型，以及 (ii) ATF4/CHOP 如何调节 黑色素瘤转移与耐药性？我们的研究将依赖于同类的黑色素瘤，无论是幼稚的还是 对治疗有抵抗力的同源 Braf/Pten 衍生黑色素瘤系 CRSPER'd 的 UPR 和 Siah1is2 基因，以及 遗传黑色素瘤模型与 Atf4、Chop 或 Siah1 突变动物杂交。我们的研究预计 带来前所未有的新见解，了解选择 UPR 组件在控制中所发挥的精确作用 黑色素瘤的可塑性，是其耐药性和转移表型倾向的基础—— 代表了一个关键的未满足的临床需求。