Multiplex approach to DNA methylation assays in exhaled breath condensate

呼出气冷凝液中 DNA 甲基化检测的多重方法

• 批准号: 7648269
• 负责人: WEIGUO HAN
• 金额: $ 8.3万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7648269
• 关键词: Age-Years; Alleles; Base Sequence; Biological Assay; Biological Markers; CDH1 gene; Chromosome Mapping; Clinical Research; Communities; DNA; DNA Methylation; DNA analysis; Development; Diagnostic; Dose; Exhalation; Future; Genes; Genome; Genomics; Goals; High-Risk Cancer; Hypermethylation; Incidence; Individual; Laboratories; Literature; Lung; MGMT gene; Malignant Neoplasms; Malignant neoplasm of lung; Maps; Methods; Methylation; Multivariate Analysis; Oral; Performance; Population; Prevention program; Research; Risk; Risk Assessment; Sampling; Screening procedure; Sensitivity and Specificity; Smoker; Smoking Status; Sputum; Techniques; Testing; Tobacco smoke; Tumor Suppressor Genes; Validation; Work; base; bisulfite; cancer initiation; cancer risk; improved; lung cancer screening; lung carcinogenesis; molecular marker; mortality; non-smoker; promoter; tool

DESCRIPTION (provided by applicant): The high mortality for lung cancer urgently requires the availability of new, noninvasive diagnostic tools for use in screening and prevention programs. Our long term goal is to develop screening approach for lung cancer. Silencing of tumor suppressor genes by aberrant promoter hypermethylation is recognized as a crucial component in lung cancer initiation and progression. Recently, we have been able to map gene promoter methylation in DNA samples recovered from EBC. However, the number of genes that can be analyzed in any given EBC sample is limited by the quantity of DNA template. Therefore, improvement in this method to multiplex the assay is necessary before expanding validation testing. The objective in this R03 proposal is to improve the method for multiple gene methylation analysis in exhaled breath condensate. We hypothesize that the current method can be modified by the addition of a Whole Genome Amplification (WGA) step to make it possible to perform multiple gene methylation analyses in EBC samples. To validate the utility of the developed method, we will analyze the methylation status, with and without WGA, of p16, DAPK, RASSF1A, MGMT, PAX51, PAX52 and CDH1 in sixty EBC samples from smokers and non-smokers, and also compare EBC with sputum from the same subjects, for reference. Our rationale for these studies is that their successful completion would provide the research community with a new, non-invasive method for further clinical studies that evaluate DNA methylation as a biomarker for risk assessment, or early detection, of lung cancer. Project Narrative: The high mortality from lung cancer warrants the development of new, noninvasive diagnostic tools for use in screening and prevention programs. Recently, we have been able to assess gene promoter methylation, a cancer biomarker, in DNA samples recovered from exhaled breath. The objective in this R03 proposal is to improve the method for multiple gene methylation analysis in exhaled breath condensate, to provide the research community with a new, non-invasive method for assessing an individual's risk for lung cancer.

描述（由申请人提供）：急性肺癌死亡率高死亡率，需要在筛查和预防计划中使用新的无创诊断工具。我们的长期目标是开发肺癌筛查方法。通过异常启动子高甲基化对肿瘤抑制基因的沉默被认为是肺癌开始和进展中的关键成分。最近，我们能够在从EBC中回收的DNA样品中绘制基因启动子甲基化。但是，可以在任何给定的EBC样品中分析的基因数量受DNA模板的量限制。因此，在扩展验证测试之前，需要改进此方法以多重分析。该R03提案的目的是改善呼出呼吸凝结物中多种基因甲基化分析的方法。我们假设可以通过添加整个基因组扩增（WGA）步骤来修改当前方法，以使在EBC样品中执行多个基因甲基化分析成为可能。为了验证开发方法的实用性，我们将分析p16，DAPK，RASSF1A，MGMT，MGMT，PAX51，PAX51，PAX51和CDH1的甲基化状态在吸烟者和非烟民中的60个EBC样品中，还将EBC与来自同一受试者的Sputum进行比较。我们对这些研究的理由是，它们的成功完成将为研究界提供一种新的，无创的方法，用于进一步的临床研究，以评估DNA甲基化作为肺癌风险评估或早期检测的生物标志物。项目叙述：肺癌的高死亡率值得开发用于筛查和预防计划的新的无创诊断工具。最近，我们能够评估从呼出的呼吸中回收的DNA样品中的一种癌症生物标志物基因启动子甲基化。该R03提案的目的是改善呼出呼吸凝结物中多种基因甲基化分析的方法，为研究界提供一种新的无创方法，以评估个人的肺癌风险。