Screening DNA Methylation in Bodily Fluids for Early Cancer Diagnostics

筛查体液中的 DNA 甲基化以进行早期癌症诊断

基本信息

批准号：
8320756
负责人：
Tza-Huei Jeff Wang
金额：
$ 119.36万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8320756
关键词：
Biological Assay Biological Markers Biosensing Techniques Blood Body Fluids Cancer Center Cancer Control Cancer Detection Cancer Diagnostics Cancer Patient Chemicals Clinical Colon Colon Carcinoma DNA DNA Methylation Detection Development Early Diagnosis Epigenetic Process Feces Fluorescence Resonance Energy Transfer Gene Silencing Goals Hour Hypermethylation Individual Liquid substance Lung Neoplasms Malignant Neoplasms Malignant neoplasm of lung Manuals Mediating Methods Methylation Molecular Monitor Mutation Nanotechnology Noise Nucleic Acids Patients Phase Plasma Preparation Process Quantum Dots Reaction Recovery Research Personnel Sampling Screening for cancer Screening procedure Sensitivity and Specificity Signal Transduction Silicon Dioxide Solid Sputum Staging System Techniques Technology Testing Therapeutic Time Tube bisulfite cancer diagnosis design high risk high throughput screening improved miniaturize particle promoter response tumor tumor progression

项目摘要

The most studied epigenetic abnormality in cancer is gene silencing associated with DNA hypermethylation. Recent studies demonstrate that gene inactivation by promoter hypermethylation can occur at early stages of cancer progression, perhaps even before mutations can be detected. Using DNA methylation as a cancer biomarker shows great promise for early diagnosis, assessments in high risk individuals, and post-therapy monitoring. While a noninvasive test with bodily fluids to detect cancer is seen as a holy grail by clinicians, the ability to detect specific DNA methylation changes in bodily fluids, including blood, sputum or stool represents a greater challenge, due to the small amounts of DNA available in these samples and the limited tumor content of such samples. Therefore, a clinically useful technology that allows for detection of DNA methylation in bodily fluids will have a substantial impact in both cancer diagnosis and management. We propose to develop a new methylation detection platform which integrates improved methods for each of the critical processes involved: DNA isolation, bisulfite treatment, and detection of methylation. All steps will ultimately be integrated and performed on a microfiuidic chip, utilizing superparamagnetic particles as a common carrier for fluidic and molecular manipulations and the quantum dots-mediated fluorescence resonance energy transfer technology (QD-FRET) for biosensing. This approach will facilitate highly efficient sample preparation and sensitive detection of DNA methylation in bodily fluids. In addition, the proposed technology can perform integrated and automatic analysis, minimizing manual labor and time while providing more reproducible results and being useful for a large scale screening. The potential use in pre-cUnical applications will be determined by testing bodily samples from patients with early stage cancers and controls, including serum/plasma, sputum and stool. In addition, the detection of cancer specific methylation in the blood/plasma will be used to monitor therapeutic response in Projects 2, 3 and 4, facihtating development prior to examination of cancer patients.

癌症中研究最多的表观遗传异常是与 DNA 高甲基化相关的基因沉默。最近的研究表明，启动子高甲基化导致的基因失活可能发生在癌症进展的早期阶段，甚至可能在检测到突变之前发生。使用 DNA 甲基化作为癌症生物标志物在早期诊断、高危个体评估和治疗后监测方面显示出巨大的前景。虽然临床医生将利用体液进行无创检测癌症视为圣杯，但由于可用的 DNA 量很少，检测体液（包括血液、痰或粪便）中特定 DNA 甲基化变化的能力是一个更大的挑战这些样品中的肿瘤含量有限。因此，一种能够检测体液中 DNA 甲基化的临床有用技术将对癌症诊断和治疗产生重大影响。我们建议开发一个新的甲基化检测平台，该平台集成了涉及的每个关键过程的改进方法：DNA分离、亚硫酸氢盐处理和甲基化检测。所有步骤最终将在微流控芯片上集成和执行，利用超顺磁性颗粒作为流体和分子操作的通用载体，以及用于生物传感的量子点介导的荧光共振能量转移技术（QD-FRET）。这种方法将有助于高效的样品制备和体液中 DNA 甲基化的灵敏检测。此外，所提出的技术可以执行集成和自动分析，最大限度地减少体力劳动和时间，同时提供更具可重复性的结果，并可用于大规模筛选。在临床前应用中的潜在用途将通过测试早期癌症患者和对照患者的身体样本（包括血清/血浆、痰和粪便）来确定。此外，血液/血浆中癌症特异性甲基化的检测将用于监测项目2、3和4中的治疗反应，促进癌症患者检查之前的开发。