Decoding AMPK-dependent regulation of DNA methylation in lung cancer

解码肺癌中 DNA 甲基化的 AMPK 依赖性调节

• 批准号: 10537799
• 负责人: Shira Yomtoubian
• 金额: $ 6.76万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-19 至 2025-06-18
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10537799
• 关键词: 5' -AMP-activated protein kinase; Address; Alanine; Binding; Bioinformatics; Biological Assay; Cancer Cell Growth; Cancer Etiology; Cancer Patient; Cancer cell line; Cell Proliferation; Cell Respiration; Cell physiology; Cells; Characteristics; Consensus; DNA Methylation; DNA Methylation Regulation; DNA Modification Methylases; Epigenetic Process; Exhibits; Face; Generations; Genetic; Genus Hippocampus; Growth; Homologous Gene; Impairment; In Vitro; KRAS oncogenesis; KRAS2 gene; KRASG12D; Knock-in; Knock-out; Lead; Lesion; Lung Neoplasms; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Metabolic; Metabolic stress; Metabolism; Methylation; Methyltransferase; Mitochondria; Modeling; Mus; Mutation; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Pathway interactions; Patients; Phenotype; Phospho-Specific Antibodies; Phosphorylation; Protein Kinase; Proteins; Rattus; Refractory; Regulation; Reporting; Role; Serine; Signal Pathway; Signal Transduction; Testing; Time; Tumor Burden; Tumor Suppressor Proteins; Tumorigenicity; Validation; Viral Oncogene; Work; base; bisulfite sequencing; cancer cell; epigenome; flexibility; improved; in vivo; insight; lung cancer cell; lung colonization; methylation pattern; migration; mortality; mouse model; mutant; neoplastic cell; novel; programs; sarcoma; sensor; targeted treatment; therapeutic target; therapeutically effective; transcriptome sequencing; tumor; tumor growth; tumor initiation; tumor progression; tumorigenic; whole genome

Project Summary Lung cancer is the leading cause of cancer-related mortality worldwide. Lung tumors driven by mutant KRAS are among the most aggressive and refractory to treatment, due in part by KRAS-driven metabolic reprogramming. Efforts to better understand the pathways regulating metabolic adaptations in oncogenic KRAS- driven tumors will provide insight into lung cancer progression and identify vulnerabilities that could be therapeutically targeted to improve patient survival. The Shaw lab recently showed the requirement of AMP- activated protein kinase (AMPK) to promote the growth of oncogenic KRAS-driven non-small-cell lung cancer (NSCLC). AMPK is a master regulator of cellular and organismal metabolism that acts as a sensor of cellular energy by altering metabolism when energy levels are low. While the Shaw lab and others have demonstrated that AMPK signaling provides cancer cells with flexibility to adapt to metabolic stresses, the epigenetic mechanisms by which AMPK promotes metabolic alterations and lung tumor growth remain poorly understood. Preliminary studies identified a de novo DNA methyltransferase as a novel substrate of AMPK. DNA methylation is involved in many normal cellular processes and is abnormally distributed in cancer cells, contributing to some of their aggressive characteristics. This proposal addresses the consequences of AMPK-dependent regulation of the de novo DNA methyltransferase on DNA methylation, metabolic programs, and lung tumor growth. First, this work aims to define the methylation profile controlled by the AMPK-dependent phosphorylated form of the de novo DNA methyltransferase using whole-genome bisulfite sequencing, CUT&TAG, and RNA sequencing assays. Tumorigenicity and Seahorse real-time cell metabolic analyses will determine whether this regulation disrupts DNA methylation patterns in a manner that generates tumor-promoting epigenetic lesions and metabolic alterations. Additionally, the generation of autochthonous KRAS-driven NSCLC mouse lines expressing constitutive knock-in of the de novo DNA methyltransferase with a serine-to-alanine mutation at the putative phosphor-acceptor-serine will enable testing whether regulation of the de novo DNA methyltransferase impacts tumor initiation, growth, and metastasis. This work addresses a fundamental relationship between two hallmarks of cancer and if successful would lead to the mechanistic connection between metabolic stresses tumor cells face and how they may trigger sustained DNA methylation changes.

项目摘要 肺癌是全球与癌症相关死亡率的主要原因。突变KRAS驱动的肺肿瘤 是最具侵略性和难治性的治疗之一，部分由KRAS驱动的代谢 重新编程。努力更好地理解调节致癌kras的代谢适应的途径 驱动的肿瘤将提供对肺癌进展的见解，并确定可能是 治疗目标以改善患者的生存。 Shaw Lab最近显示了AMP-的要求 活化的蛋白激酶（AMPK）促进致癌KRAS驱动的非小细胞肺癌的生长 （NSCLC）。 AMPK是细胞和生物代谢的主要调节剂，充当细胞的传感器 当能量水平较低时，能量通过改变代谢来改变。而肖实验室和其他实验室已经证明了 AMPK信号传导可为癌细胞具有适应代谢应力的灵活性，即表观遗传学 AMPK促进代谢改变和肺部肿瘤生长的机制仍然很少了解。 初步研究确定了从头DNA甲基转移酶是AMPK的新底物。 DNA甲基化 参与许多正常细胞过程，并异常分布在癌细胞中，有助于某些 它们的积极特征。该提案解决了依赖AMPK的法规的后果 DNA甲基转移酶在DNA甲基化，代谢程序和肺部肿瘤生长上。第一的， 这项工作旨在定义由由AMPK依赖性磷酸化形式控制的甲基化剖面 使用全基因组亚硫酸盐测序，切割和TAG和RNA测序的从头DNA甲基转移酶 测定。肿瘤性和海马实时细胞代谢分析将确定该调节是否是否 以产生促肿瘤表观遗传病变和代谢的方式破坏DNA甲基化模式 改变。此外，表达的自动量KRAS驱动的NSCLC鼠标线的产生 在推定的丝氨酸到丙氨酸突变的构成型DNA甲基转移酶 磷光受感受器 - 丝氨酸将能够测试从头DNA甲基转移酶的调节是否影响 肿瘤起始，生长和转移。这项工作解决了两个标志之间的基本关系 癌症和如果成功的话会导致代谢应力之间的机械联系，肿瘤细胞 面部以及它们如何触发持续的DNA甲基化变化。