CIRCULATING DNA AMPLIFICATION & COLON CA DETECTION

循环 DNA 扩增

• 批准号: 7090955
• 负责人: G. Mike Makrigiorgos
• 金额: $ 19.5万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-08 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7090955
• 关键词: DNA; clinical research; colon; genome; neoplasm /cancer; plasma

DESCRIPTION (provided by applicant): This 2-year R21 project will demonstrate that newly developed genome amplification and analysis technologies lead to highly improved sensitivity in detecting circulating nucleic acids of tumor origin in the plasma of patients with cancer. In particular, we have developed a new technology, Restriction and Circularization-Aided Rolling Circle Amplification, RCA-RCA that allows comprehensive whole genome amplification from DNA that has undergone fragmentation (i.e. the method is tolerant to sample degradation). As such, it can amplify fragmented nucleic acids circulating in human plasma, including tumor-derived DNA that is used as a biomarker for early tumor detection and disease monitoring in cancer. In a modified protocol, RCA-RCA can also achieve whole methylome amplification, i.e., it preserves epigenetic changes so that methylation can be studied on a high throughput basis in the amplified material. We aim to demonstrate that whole genome/methylome amplification enables highly improved detection of tumor-derived genetic or epigenetic alterations in blood plasma by providing unlimited material for high- throughput molecular analysis and by allowing high sensitivity and identification of new biomarkers via microarray analysis of plasma-circulating DNA. The proposed study will focus on colon cancer. The premise of this proposal is that, in early colon cancer, tumor DNA is circulating in the plasma but often goes undetected due to limitations in the detection method(s) and the material available. By enabling 'target magnification' and application of high-throughput genome analysis we aim to achieve highly improved sensitivity and reliability in detecting circulating nucleic acids of tumor origin in plasma. Furthermore, the present whole genome amplification enables plasma-DNA archives to be established that enable future retrospective studies to be conducted on the same samples for evaluation of new biomarkers and sharing of material among investigators.

描述（由申请人提供）：这个为期 2 年的 R21 项目将证明，新开发的基因组扩增和分析技术可显着提高检测癌症患者血浆中肿瘤来源的循环核酸的灵敏度。特别是，我们开发了一项新技术，即限制性和环化辅助滚环扩增（RCA-RCA），它允许从经历片段化的 DNA 中进行全面的全基因组扩增（即该方法能够耐受样品降解）。因此，它可以扩增人类血浆中循环的片段化核酸，包括肿瘤衍生的 DNA，用作早期肿瘤检测和癌症疾病监测的生物标志物。在修改后的方案中，RCA-RCA还可以实现全甲基化组扩增，即，它保留表观遗传变化，以便可以在扩增材料中以高通量为基础研究甲基化。我们的目标是证明全基因组/甲基化组扩增能够为高通量分子分析提供无限的材料，并通过血浆循环 DNA 的微阵列分析实现新生物标志物的高灵敏度和鉴定，从而能够显着改善血浆中肿瘤源性遗传或表观遗传改变的检测。拟议的研究将重点关注结肠癌。该提议的前提是，在早期结肠癌中，肿瘤 DNA 在血浆中循环，但由于检测方法和可用材料的限制，常常未被检测到。通过实现“目标放大”和高通量基因组分析的应用，我们的目标是在检测血浆中肿瘤来源的循环核酸方面实现显着提高的灵敏度和可靠性。此外，目前的全基因组扩增能够建立血浆 DNA 档案，从而能够在未来对相同样本进行回顾性研究，以评估新的生物标志物并在研究人员之间共享材料。