Array Based CGH for Genome-Wide Analysis of Wilms' Tumor

基于阵列的 CGH 用于肾母细胞瘤全基因组分析

• 批准号: 6560241
• 负责人: Norma Jean Nowak
• 金额: $ 15.51万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-10 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6560241
• 关键词: Wilms' tumor; artificial chromosomes; cell differentiation; chromosome aberrations; chromosome deletion; comparative genomic hybridization; developmental genetics; high throughput technology; human genetic material tag; human tissue; loss of heterozygosity; microarray technology; molecular cloning; neoplasm /cancer genetics; neoplastic transformation; nephrogenesis; nucleic acid hybridization; technology /technique development

DESCRIPTION (provided by applicant): Wilms' tumor (WT) is the most common primary malignant solid tumor of childhood, accounts for 6-7% of all childhood malignancies in the United States, and is an important model for the study of the fundamental mechanisms of tumorigenesis. The WT1 gene was identified and positionally cloned in 1991, but we have shown that this gene is only involved in a very small percentage (-5-7%) of WT patients. Since the cloning of WT1, no additional genes have been shown to be directly involved in the development of WT. Several groups, including our own, have attempted to identify new regions associated with WT through genome wide LOH (Loss of Heterozygosity) analysis and CGH (Comparative Genomic Hybridization) analysis on metaphase chromosomes, resulting in the association of chromosomes 5q (22%) 7p (15%) and 16q (12%) with WT. While these studies have provided some evidence for genomic regions harboring genes involved in Wilms' tumorigenesis, no gene has yet been cloned to coincide with these regions. These efforts have been hampered by the lack of robust, high throughput, high resolution techniques for detecting submicroscopic deletions and amplifications. Thus, the genetic defect(s) in the majority of these patients still remains unclear. It is clear that WT results from abrogation of the normal developmental pathways within the kidney. Identification of additional genes involved in WT pathogenesis will not only advance our understanding of Wilms' tumorigenesis, but also uncover crucial events required for proper cellular differentiation and normal kidney development. We have been developing a high throughput array based technology that will allow us to scan the genome for deletions and amplifications, or copy number aberrations (CNAs), at 0.5-1mb level of resolution. Hybridization of tumor DNA to these arrays readily identifies losses and gains in the tumor samples using a normal DNA reference. We will identify the recurrent copy number aberrations present in our WT bank, tile the implicated chromosomal regions on WT subarrays, and identify candidate genes involved in WT and kidney development.

描述（由申请人提供）：Wilms的肿瘤（WT）是最常见的原发性恶性实体瘤，占美国所有儿童恶性肿瘤的6-7％，并且是研究肿瘤发生基本机制的重要模型。 WT1基因在1991年被鉴定出来并在位置克隆，但我们已经表明，该基因仅涉及少数（-5-7％）的WT患者。自WT1的克隆以来，尚未证明其他基因直接参与WT的发展。包括我们自己在内的几个小组试图通过基因组宽LOH（杂合性的丧失）分析和CGH（比较基因组杂交）分析中期染色体，从而鉴定与WT相关的新区域，从而导致染色体5q（22％）7p（15％）和16Q（12％）和WT TT的染色体相关。尽管这些研究提供了一些基因组区域的证据，这些基因组含有Wilms肿瘤的基因，但尚未克隆基因与这些区域一致。缺乏可检测亚镜下缺失和放大的可靠，高吞吐量，高分辨率技术的努力。因此，大多数患者的遗传缺陷仍然不清楚。显然，WT是由于废除了肾脏中正常发育途径而引起的。鉴定参与WT发病机理的其他基因不仅会提高我们对Wilms肿瘤发生的理解，而且还会发现适当的细胞分化和正常肾脏发育所需的关键事件。我们一直在开发高吞吐量阵列的技术，该技术将使我们能够以0.5-1MB的分辨率扫描基因组中的删除和放大或复制号畸变（CNAS）。肿瘤DNA与这些阵列的杂交很容易通过正常的DNA参考来鉴定肿瘤样品中的损失和收益。我们将确定我们WT银行中存在的复制拷贝数畸变，瓷砖与WT子阵列有关的染色体区域，并确定与WT和肾脏开发有关的候选基因。