Role of the DREAM complex in the lung tumor suppression

DREAM 复合物在抑制肺部肿瘤中的作用

基本信息

批准号：
10575588
负责人：
Larisa Litovchick
金额：
$ 21.77万
依托单位：
VIRGINIA COMMONWEALTH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-03 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10575588
关键词：
Address Adenocarcinoma Cell Alleles Animals Binding Breeding Bypass CDKN2A gene CRISPR/Cas technology Cancer Etiology Cell Aging Cell Cycle Cell Cycle Arrest Cell Cycle Progression Cell Proliferation Cell Survival Cells Cessation of life Chronic Clinical Data Communities Complex Copy Number Polymorphism DNA Damage DNA Repair DNA Repair Gene Data Analyses Detection Development Diagnostic Disease Early Diagnosis Environmental Risk Factor Epithelial Cell Proliferation Epithelial Cells Experimental Models Exposure to Future Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Genetic Transcription Genetically Engineered Mouse Genome Genotoxic Stress Growth Growth Factor Heterozygote Histologic Human Human Cell Line Immune In Vitro Inflammatory Investigation KRAS2 gene Knock-in Knowledge Lesion Lung Lung Adenocarcinoma Lung Neoplasms Malignant - descriptor Malignant Neoplasms Malignant neoplasm of lung Measurement Mediating Messenger RNA Modeling Molecular Mouse Cell Line Mus Mutant Strains Mice Mutate Mutation Non-Small-Cell Lung Carcinoma Oncogenic Outcome Pathogenesis Pathway interactions Phenotype Phosphorylation Phosphotransferases Physiological Play Prevention Prevention strategy Proliferating Proliferation Marker Protein Family Protein Kinase Proteins Radiation Radiation induced damage Radon Recording of previous events Repression Repressor Proteins Research Retinoblastoma Retinoblastoma Protein Risk Factors Role Serine Signal Transduction Smoker Squamous cell carcinoma Staging TP53 gene Testing The Cancer Genome Atlas Tobacco smoke Tobacco smoking behavior Transcription Repressor Tumor Suppression Tumor Suppressor Proteins airway epithelium cancer initiation cdc Genes cell injury detection of nutrient driver mutation genome editing genotoxicity high reward high risk irradiation lung carcinogenesis lung histology lung tumorigenesis mRNA Expression mouse model mutant neoplastic novel novel strategies premalignant prevent promoter protein expression recruit repaired response senescence small cell lung carcinoma survival outcome targeted treatment tobacco exposure tumor tumor progression tumorigenesis

项目摘要

ABSTRACT Lung cancer is the major cause of cancer deaths in the U.S. and worldwide. The most common types of lung cancer are non-small cell lung carcinoma (NSCLC, 84%) and small cell lung carcinoma (SCLC, 13%). NSCLC includes two major histological subtypes, adenocarcinoma (AC), and squamous cell carcinoma (SCC). All subtypes of lung cancer are thought to arise from airway epithelial cells damaged by genotoxic exposure, which most commonly include tobacco smoke and radon radiation. These damaged cells frequently acquire early driver mutations in the KRAS gene that could be also present in advanced lung cancers. However, even in the cases with significant and chronic exposure (such as in heavy smokers), not all damaged cells give rise to cancer, and the mechanisms that prevent the progression of early premalignant lesions to cancer are still not fully understood. Studies using genetically engineered mouse models (GEMMs) highlighted the roles of cell cycle arrest and senescence, mediated by retinoblastoma (Rb) family proteins and p53, in protection from lung tumorigenesis. However, further research is required to better understand the barriers to lung carcinogenesis at the molecular level, and to develop accurate models representing early stages of lung cancer pathogenesis. Previously, we characterized a transcriptional repressor complex called DREAM that assembles in G0/G1 when Rb-like protein p130 recruits E2F4, DP1, and a stable core of five proteins including LIN9, LIN37, LIN52, LIN54, and RBBP4. The DREAM binds to promoters and mediates the repression of >800 cell cycle genes, including most genes required for DNA damage response and repair. Phosphorylation of serine-28 residue in LIN52 by DYRK1A kinase is required for the DREAM assembly. DREAM-DYRK1A pathway could be relevant to cancer because either inhibition of DYRK1A, or S28A-LIN52 mutation result in bypass of the oncogenic Ras- induced senescence in the immortalized human and mouse cell lines. The KRAS gene is mutated in 15% - 30% of NSCLC cases, and its driver role in lung cancer is strongly supported by encouraging NSCLC clinical data using KRASG12C-targeted therapeutics. However, the pathophysiological significance of the DREAM disruption in lung cancer is not known, and the mechanisms of senescence bypass in the cells lacking DREAM are not well understood. Here, we test our hypothesis that DREAM complex contributes to tumor prevention by suppressing DNA repair in the damaged cells during early stages of lung cancer pathogenesis. Previously, the role of DREAM in lung cancer could not be directly investigated due to a lack of suitable mouse models. To address this knowledge gap, we generated a DREAM-less mouse homozygous for S28A mutation in the Lin52 gene, using CRISPR-Cas9 gene editing. We will use our novel Lin52S28A mouse model to investigate the role of the DYRK1A-DREAM pathway during the early steps of lung cancer pathogenesis driven by genotoxic stress or KrasG12C mutation.

抽象的肺癌是美国和全世界癌症死亡的主要原因。最常见的肺类型癌症包括非小细胞肺癌（NSCLC，84%）和小细胞肺癌（SCLC，13%）。非小细胞肺癌包括两种主要的组织学亚型：腺癌（AC）和鳞状细胞癌（SCC）。全部肺癌的亚型被认为是由因基因毒性暴露而受损的气道上皮细胞引起的，最常见的是烟草烟雾和氡辐射。这些受损细胞经常获得早期驱动程序 KRAS 基因突变也可能存在于晚期肺癌中。然而，即使在这些情况下如果长期大量接触（例如重度吸烟者），并非所有受损细胞都会引发癌症，并且阻止早期癌前病变进展为癌症的机制尚未完全清楚。使用基因工程小鼠模型（GEMM）的研究强调了细胞周期停滞和衰老由视网膜母细胞瘤 (Rb) 家族蛋白和 p53 介导，可防止肺部肿瘤发生。然而，还需要进一步的研究来更好地了解分子水平上肺癌发生的障碍。水平，并开发代表肺癌发病机制早期阶段的准确模型。之前，我们表征了一种名为 DREAM 的转录抑制复合物，它在 G0/G1 中组装当 Rb 样蛋白 p130 招募 E2F4、DP1 和 LIN9、LIN37、LIN52 等五种蛋白的稳定核心时， LIN54 和 RBBP4。 DREAM 与启动子结合并介导超过 800 个细胞周期基因的抑制，包括 DNA 损伤反应和修复所需的大多数基因。丝氨酸 28 残基的磷酸化 DREAM 组装需要 DYRK1A 激酶的 LIN52。 DREAM-DYRK1A 通路可能与癌症，因为 DYRK1A 或 S28A-LIN52 突变的抑制会导致致癌 Ras- 的旁路诱导永生化人类和小鼠细胞系的衰老。 KRAS基因在15% - 30%中发生突变 NSCLC 病例的数量，其在肺癌中的驱动作用得到了令人鼓舞的 NSCLC 临床数据的有力支持使用 KRASG12C 靶向疗法。然而，DREAM 破坏的病理生理学意义在肺癌中的作用尚不清楚，并且缺乏 DREAM 的细胞中衰老旁路的机制尚不清楚明白了。在这里，我们通过以下方式检验了 DREAM 复合物有助于肿瘤预防的假设：抑制肺癌发病早期阶段受损细胞的 DNA 修复。此前，由于缺乏合适的小鼠，无法直接研究DREAM在肺癌中的作用模型。为了解决这一知识差距，我们生成了 S28A 突变的无 DREAM 纯合小鼠在 Lin52 基因中，使用 CRISPR-Cas9 基因编辑。我们将使用我们新颖的 Lin52S28A 小鼠模型进行研究 DYRK1A-DREAM 通路在基因毒性驱动的肺癌发病机制早期阶段中的作用压力或 KrasG12C 突变。