High-Resolution CGH Charaterization of Brain Tumors

脑肿瘤的高分辨率 CGH 表征

• 批准号: 6884670
• 负责人: SIMON G GREGORY
• 金额: $ 17.81万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-15 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6884670
• 关键词: artificial chromosomes; astrocytoma; biotechnology; brain disorder diagnosis; brain neoplasms; chromosome deletion; chromosome translocation; clinical research; comparative genomic hybridization; diagnosis design /evaluation; genetic mapping; genetic markers; glioblastoma multiforme; human tissue; microarray technology; neoplasm /cancer diagnosis; neoplasm /cancer genetics; neoplastic growth; nucleic acid amplification techniques; nucleic acid purification; oligodendroglia; oligonucleotides; polymerase chain reaction; prognosis; tissue /cell culture

DESCRIPTION (provided by applicant): It has been known for many years that malignant tumors have chromosome losses that contribute to their malignancy. In recent years, considerable research effort has been directed at identifying the one common region of deletion that occurs within specific tumor types in order to identify the actual gene(s) involved in the transformations. Traditionally, loss of heterozygosity mapping, using polymorphic markers and genotype information, and comparative genomic hybridization (CGH), utilizing normal metaphase chromosomes as the template upon which differentially labeled test and control samples are hybridized, have been used to identify chromosomal rearrangements. However, these approaches have proven to be relatively time consuming and have low resolution when identifying aberrant chromosomal regions. The project outlined here describes the development of high-resolution, chromosome specific, CGH arrays for the detection of chromosomal deletions and amplifications in neurological neoplasia. High-resolution array CGH will allow for very rapid and accurate (100 -200 kilobase) characterization of chromosomal rearrangements within a large number of tumors. High-resolution genome-wide coverage, on a per-chromosome basis, will be achieved by utilizing complete, overlapping, minimum-tile path clones that were generated by the Human Genome Project in its production of highly accurate genomic sequence. The research project contains four basic aims; aim 1, the generation of genome-wide 1Mb, chromosome and region specific high-resolution CGH arrays, including the purification of minimum-tile path DNA and production of amino linked DOP-PCR products; aim 2, the characterization of the arrays by testing with differentially labeled normal DNA's as well as known deletions and amplifications; aim 3, the characterization of clinically defined brain tumor types (glioblastoma multiforme, oligoastrocytoma and oligodendroglioma) by high-resolution CGH hybridization to determine exact regions of chromosomal rearrangement and therefore the putative disease causing genes within them; aim 4, investigate the use of high-resolution CGH arrays as a diagnostic tool for predicting the responsiveness of oligodendrogliomas to chemotherapeutic treatment that are associated with known chromosome deletions.

描述（由申请人提供）：多年来一直知道恶性肿瘤的染色体损失会导致其恶性肿瘤。近年来，已经大量的研究工作旨在确定特定肿瘤类型中发生的一个共同缺失区域，以识别转化涉及的实际基因。传统上，使用多态性标记和基因型信息以及比较基因组杂交（CGH）的杂合性映射的丢失，利用正常的中期染色体作为模板，用于鉴定染色体的测试和对照样品被杂交的模板。但是，这些方法已被证明是相对耗时的，并且在识别异常染色体区域时的分辨率很低。 这里概述的项目描述了用于检测神经系统肿瘤中染色体缺失和扩增的高分辨率，特异性CGH阵列的发展。高分辨率阵列CGH将允许在大量肿瘤中非常快速准确（100 -200千倍）对染色体重排的表征。以每个染色体为基础，将通过使用人类基因组项目产生的完整，重叠的，最小瓷砖路径克隆来实现高分辨率全基因组的覆盖范围，以实现其高度准确的基因组序列。 研究项目包含四个基本目标。 AIM 1，全基因组1MB，染色体和区域特异性高分辨率CGH阵列的产生，包括纯化最小瓷砖路径DNA和氨基连接的DOP-PCR产品的产生； AIM 2，通过用差异标记的正常DNA以及已知的缺失和扩增来测试阵列的表征； AIM 3，通过高分辨率CGH杂交来表征临床定义的脑肿瘤类型（多形胶质细胞瘤，多形拟南芥和少突胶质瘤），以确定染色体重排的确切区域，从而确定其内基因的假定疾病。 AIM 4，研究使用高分辨率CGH阵列作为一种诊断工具，以预测与已知染色体缺失相关的化学治疗治疗的少突胶质瘤的反应。