The Oncogene Activated Mitochondrial Unfolded Protein Response Regulates Senescence Biology

项目总结 线粒体为真核生物和后生动物组织创造了进化优势，以及它们对 细胞生物学从合成代谢和分解代谢延伸到决定细胞存活的最后时刻 参与细胞凋亡。在过去的十年里，人们对研究线粒体如何影响癌细胞的兴趣 生物学使我们的实验室确定了将致癌信号(即BRAFV600E/NRASG12V)与 恶性细胞内多个以线粒体为中心的过程，包括改变的线粒体动力学， 氧化磷酸化和化疗敏感性。最近，我们专注于探索癌基因是如何 在转化之前与线粒体生物学相交-这可能会提供前- 恶性细胞生物学和疾病早期阶段。我们通常把我们的研究放在黑色素瘤的背景下 由于我们在原代人类黑素细胞方面有丰富的经验，患者rna-seq的整合队列 数据集和组织，以及早期和晚期疾病的多种体外和体内模型。例如， 在原代人黑素细胞中引入致癌信号(BRAFV600E/NRASG12/Q60)导致快速 癌基因诱导衰老(OIS)，这种肿瘤抑制机制反映在 在临床上表现为皮肤癌前病变。因此，这一应用的科学前提是基于 在三个新的观察上：(I)表达癌基因的原代人类黑素细胞迅速扩大其 OIS期间的线粒体网络；(Ii)这种扩张是由ATF5- 依赖线粒体未折叠蛋白反应(MtUPR)；和(Iii)mtUPR控制 奥伊斯。虽然mtUPR是细胞器特有的基本质量控制信号通路，但它对 缓解线粒体应激，没有文献机械地将致癌信号与mtUPR激活联系起来， 黑素细胞生物学，也不是黑色素瘤进展。在我们的初步实验中，我们探索了生化 信号、线粒体反应、细胞功能获得/功能丧失途径，以及数百种 患者样本建立癌基因激活的ATF5依赖的mtUPR是关键的假设 在OIS及其逃逸过程中指导黑素细胞的信号通路。在此R01应用程序中，我们建议 三个互补的，但不同的，具体的目的是使用黑素细胞，无数的模型来检验这一假说 早期疾病和病人样本。具体目标#1：审问 致癌信号、mtUPR和OIS。具体目标2：确定基因表达程序介导的和 在OIS和早期疾病原发黑色素瘤期间由mtUPR维持。具体目标3：确定影响 在色素痣和原发病模型中mtUPR的激活。

PROJECT SUMMARY Mitochondria created an evolutionary advantage for eukaryote and metazoan organization, and their impact on cell biology extends from anabolic and catabolic metabolism to determining the final moments of cell survival by engaging apoptosis. Throughout the last decade, interest in studying how mitochondria influence cancer cell biology led our laboratories to identify mechanisms linking oncogenic signaling (i.e., BRAFV600E / NRASG12V ) to multiple mitochondria-centric processes within malignant cells including altered mitochondrial dynamics, oxidative phosphorylation, and chemosensitivity. More recently, we focused on exploring how oncogenes intersect upon mitochondrial biology prior to transformation – which will likely provide molecular details into pre- malignant cell biology and early stages of disease. We commonly position our studies in the context of melanoma as we have extensive experience with primary human melanocytes, integrated cohorts of patient RNA-seq datasets and tissues, and multiple in vitro and in vivo models of early and late disease. For instance, the introduction of oncogenic signaling (BRAFV600E / NRASG12/Q60) in primary human melanocytes causes rapid oncogene-induced senescence (OIS), and this tumor-suppressive mechanism is reflected in patients who present with pre-malignant skin lesions in the clinic. Therefore, the scientific premise for this application is based on three novel observations: (i) primary human melanocytes expressing oncogenes rapidly expand their mitochondrial networks during OIS; (ii) this expansion is dictated by the undescribed activation of the ATF5- dependent mitochondrial unfolded protein response (mtUPR); and (iii) the mtUPR controls the rate and extent of OIS. While the mtUPR is a fundamental organelle-specific quality control signaling pathway that is essential to mitigate mitochondrial stress, no literature mechanistically connects oncogenic signaling to mtUPR activation, melanocyte biology, nor melanoma progression. In our preliminary experiments, we explored biochemical signaling, mitochondrial responses, cellular gain-of-function / loss-of-function approaches, and hundreds of patient samples to establish the hypothesis that the oncogene-activated ATF5-dependent mtUPR is a key signaling pathway that instructs melanocytes during OIS and its escape. In this R01 application, we propose three complimentary, but distinct, specific aims to examine this hypothesis using melanocytes, numerous models of early disease, and patient samples. Specific Aim #1: Interrogate the mechanistic relationship between oncogenic signaling, mtUPR, and OIS. Specific Aim #2: Identify the gene expression programs mediated and maintained by the mtUPR during OIS and early disease primary melanoma. Specific Aim #3: Define the impact of mtUPR activation in models of nevi and primary disease.