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: 6746976
• 负责人: Norma Jean Nowak
• 金额: $ 15.67万
• 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 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6746976
• 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（申请人提供）：肾母细胞瘤（WT）是儿童期最常见的原发性恶性实体瘤，占美国所有儿童期恶性肿瘤的6-7%，是基础研究的重要模型。肿瘤发生机制。 WT1基因于1991年被鉴定并定位克隆，但我们已经证明该基因仅涉及极小比例（-5-7%）的WT患者。自从克隆 WT1 以来，没有其他基因被证明直接参与 WT 的发育。包括我们自己在内的几个小组已尝试通过对中期染色体进行全基因组 LOH（杂合性丢失）分析和 CGH（比较基因组杂交）分析来识别与 WT 相关的新区域，从而导致染色体 5q (22%) 7p 的关联WT (15%) 和 16q (12%)。虽然这些研究为基因组区域提供了一些证据，证明基因组区域含有与维尔姆斯肿瘤发生有关的基因，但尚未克隆出与这些区域相符的基因。由于缺乏用于检测亚显微缺失和扩增的稳健、高通量、高分辨率技术，这些努力受到阻碍。因此，大多数患者的遗传缺陷仍不清楚。很明显，WT 是肾脏内正常发育途径被破坏的结果。鉴定与 WT 发病机制相关的其他基因不仅将增进我们对肾母细胞瘤发生的理解，而且还将揭示适当的细胞分化和正常肾脏发育所需的关键事件。我们一直在开发一种基于高通量阵列的技术，该技术将使我们能够以 0.5-1mb 的分辨率扫描基因组中的缺失和扩增或拷贝数畸变 (CNA)。肿瘤 DNA 与这些阵列的杂交可以使用正常 DNA 参考轻松识别肿瘤样本中的损失和增益。我们将鉴定 WT 库中存在的反复出现的拷贝数畸变，将相关染色体区域平铺在 WT 子阵列上，并鉴定参与 WT 和肾脏发育的候选基因。