DNA Methylation Changes During Development and Progression of Lung Adenocarcinoma

肺腺癌发生和进展过程中 DNA 甲基化的变化

基本信息

批准号：
7318570
负责人：
ITE A OFFRINGA
金额：
$ 31.2万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-01 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7318570
关键词：
Address Adenocarcinoma Agar American Anchorage-Independent Growth Apoptosis Atypical adenomatous hyperplasia Biological Biological Assay Biological Markers Bronchiolo-Alveolar Adenocarcinoma Cancer Etiology Cancer Patient Cancerous Cell Cycle Cell Line Cell Mobility Cell Proliferation Cells Cessation of life Characteristics Clinical Collaborations Collection CpG Islands DNA Methylation Data Development Disease Early Diagnosis Epigenetic Process Epithelial Cells Etiology Event Gene Expression Gene Silencing Genes Genome Genomics Goals Growth Heterogeneity Histologic Hypermethylation In Vitro Incidence Invasive Lesion Lesion Life Localized Lung Lung Adenocarcinoma Lung Neoplasms Malignant Neoplasms Malignant neoplasm of lung Measurement Measures Methylation Modeling Molecular Mucinous Nature Nude Mice Pathologic Patients Phenotype Play Polymerase Chain Reaction Positioning Attribute Premalignant Purpose RNA Interference Rate Research Personnel Reverse Transcriptase Polymerase Chain Reaction Reverse Transcription Role Sampling Small Interfering RNA Smoker Staging Structure of parenchyma of lung System Therapeutic Intervention Time Tumor-Suppressor Gene Inactivation Tumorigenicity United States Woman authority base bisulfite cancer cell cell motility cell type design drug development interest knock-down matrigel men molecular modeling prevent prognostic programs promoter research study tumor

项目摘要

DESCRIPTION (provided by applicant): Lung cancer is the leading cause of cancer death in the United States. Adenocarcinoma, the histological subtype most frequently seen in never smokers and former smokers, is now the most common type of lung cancer in men and women. The increasing incidence of lung adenocarcinoma underlines the importance of understanding the development and progression of this lethal disease. DMA hypermethylation at promoter CpG islands is a key mechanism for tumor suppressor gene inactivation in cancer. Using a unique collection of pre-invasive lesions, lung adenocarcinomas of clinical stages IA-IIIA, and control lung samples, we propose to undertake a genome-wide search for DNA methylation changes associated with adenocarcinoma development and progression. The Specific Aims of this study are: 1) To identify new epigenetic changes in lung adenocarcinoma compared to histologically normal lung by genome-wide DNA methylation profiling. We will compare early and late stage adenocarcinoma and non-cancer lung to identify concurrent changes in methylation and expression, using CpG island and expression microarrays. Select loci will be verified by bisulfite sequencing and quantitative RT-PCR. 2) To determine whether DNA methylation changes for loci identified in Aim 1 occur during development and progression of lung adenocarcinoma. Using the high throughput system MethyLight, we will compare methylation profiles in 50 archival samples each of stages IA, IB, IIA, IIB and MIA lung adenocarcinoma, in pre-invasive lesions, and in non-tumor lung. 3) To determine whether correlations exist between the methylation profiles observed in Aim 2 and patient survival. 4) To examine the biological significance of loci identfied under Aim 2 using functional experiments in vitro: forcing expression of genes or artifically silencing them by RNAi. The effects of these manipulations on cell lines representing precancerous and cancerous lesions will be evaluated by measuring proliferation rates, cell cycle distribution, cell mobility, anchorage-independent growth and the ability to form tumors in nude mice. Over 150,000 Americans die every year from lung cancer. Early detection is the best way to prevent cancer deaths. The identification of sequential epigenetic alterations that occur during lung adenocarcinoma development could provide new markers for early detection and prognostication as well as possible new targets for drug development, thereby saving or extending the lives of thousands of lung cancer patients.

描述（由申请人提供）：肺癌是美国癌症死亡的主要原因。腺癌是从不吸烟者和戒烟者中最常见的组织学亚型，现在是男性和女性中最常见的肺癌类型。肺腺癌发病率的增加凸显了了解这种致命疾病的发生和进展的重要性。启动子 CpG 岛的 DMA 高甲基化是癌症中抑癌基因失活的关键机制。利用独特的浸润前病变、临床 IA-IIIA 期肺腺癌和对照肺样本的集合，我们建议对与腺癌发生和进展相关的 DNA 甲基化变化进行全基因组搜索。本研究的具体目标是：1) 通过全基因组 DNA 甲基化分析来识别肺腺癌与组织学正常肺相比新的表观遗传变化。我们将使用 CpG 岛和表达微阵列比较早期和晚期腺癌和非癌肺，以确定甲基化和表达的并发变化。选择的位点将通过亚硫酸氢盐测序和定量 RT-PCR 进行验证。 2) 确定目标 1 中确定的基因座的 DNA 甲基化变化是否发生在肺腺癌的发生和进展过程中。使用高通量系统 MethyLight，我们将比较 IA、IB、IIA、IIB 和 MIA 期肺腺癌、浸润前病变和非肿瘤肺各 50 个存档样本的甲基化谱。 3) 确定目标 2 中观察到的甲基化谱与患者生存率之间是否存在相关性。 4) 使用体外功能实验来检查目标 2 下确定的基因座的生物学意义：强制基因表达或通过 RNAi 人为沉默它们。这些操作对代表癌前病变和癌性病变的细胞系的影响将通过测量增殖率、细胞周期分布、细胞迁移性、贴壁依赖性生长和在裸鼠中形成肿瘤的能力来评估。每年有超过 150,000 名美国人死于肺癌。早期发现是预防癌症死亡的最佳方法。鉴定肺腺癌发展过程中发生的连续表观遗传改变可以为早期检测和预测提供新的标记，并为药物开发提供可能的新靶标，从而挽救或延长数千名肺癌患者的生命。