Analysis of Expression and LOH in Trout Nephroblastomas

鳟鱼肾母细胞瘤的表达和 LOH 分析

• 批准号: 6607674
• 负责人: GARY H THORGAARD
• 金额: $ 14.5万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6607674
• 关键词: DNA methylation; Wilms' tumor; carcinogenesis; chromosomes; complementary DNA; fish; gene expression; genetic mapping; genetic markers; genetic polymorphism; genetic screening; kidney; loss of heterozygosity; nephroblastoma; polymerase chain reaction; trout /salmon; tumor suppressor genes

DESCRIPTION (provided by applicant): The goal of this project is to identify differentially expressed genes and chromosomal regions associated with nephroblastoma formation in clonal lines of rainbow trout. Studies with lower vertebrates have made important comparative contributions to the understanding of carcinogenesis, and rainbow trout are a leading lower vertebrate carcinogenesis model. Trout are uniquely suited for this study because of their large size, their propensity to develop nephroblastomas, and the ongoing development of a good comparative map in this species. Human nephroblastomas (Wilms' tumor) arise from deviations in normal differentiation of a short-lived population of embryonic kidney precursor cells. Irregularities in the functioning of the Wilms' tumor gene (WT1), which is normally involved in kidney and gonadal development and sex differentiation, appear to play a fundamental role in the formation of this cancer, while mutations affecting other characterized tumor suppressor genes are rare. The key processes in the induction of this cancer seem to lay in aberrant differentiation, possibly involving the loss of imprinting, which includes anomalies in WT1 regulation. Trout offer an opportunity to study this in a genetically defined background of clonally derived populations. Nephroblastomas can be readily induced at low cost in this species, in this case by MNNG exposure in the genetically uniform hybrid progeny of crosses between two homozygous clonal trout lines. The project objectives include: 1) Analysis of gene expression differences between normal kidney and Wilms' tumor tissues by cDNA-subtraction Amplified Fragment Length Polymorphism (csAFLP). 2) We will simultaneously conduct genome-wide screens between kidney and nephroblastoma DNA using Amplified Fragment Length Polymorphism (AFLP) markers, which will reveal LOH and genomic deletions, as well as aberrant methylation. Linked markers will be used for isolating the corresponding trout BACs, which will be partially sequenced for identification of genes within the LOH deletion, allowing the determination of synteny with human chromosomes and identification of potential candidate tumor suppressor genes for further analysis. This project should provide fundamental insights into nephroblatoma carcinogenesis using an experimentally tractable lower vertebrate model system.

描述（由申请人提供）： 该项目的目标是鉴定差异表达基因并 克隆系中与肾母细胞瘤形成相关的染色体区域 虹鳟鱼。对低等脊椎动物的研究变得重要 对理解致癌作用和彩虹的比较贡献 鳟鱼是一种主要的低等脊椎动物致癌模型。鳟鱼是独一无二的 适合这项研究，因为它们的体积大，有发展的倾向 肾母细胞瘤，以及正在开发的良好比较图 物种。人类肾母细胞瘤（维尔姆斯瘤）由正常细胞的偏差引起 胚胎肾前体的短命群体的分化 细胞。维尔姆斯肿瘤基因 (WT1) 功能异常， 通常参与肾脏和性腺的发育以及性 的分化，似乎在这种现象的形成中发挥着根本性的作用。 癌症，而影响其他特征肿瘤抑制基因的突变 很少见。诱发这种癌症的关键过程似乎在于 异常分化，可能涉及印记丢失， 包括 WT1 调节异常。鳟鱼提供了研究这个的机会 在克隆衍生群体的遗传定义背景中。 在该物种中，肾母细胞瘤可以很容易地以低成本诱导，在这种情况下 MNNG 暴露在遗传均一的杂交后代中的案例 两个纯合克隆鳟鱼系之间。项目目标包括：1) 正常肾与肾母细胞瘤基因表达差异分析 通过 cDNA 消减扩增片段长度多态性 (csAFLP) 来检测组织。 2）我们将同时在肾脏和肾脏之间进行全基因组筛选 使用扩增片段长度多态性 (AFLP) 检测肾母细胞瘤 DNA 标记，这将揭示 LOH 和基因组缺失，以及异常 甲基化。链接的标记将用于隔离相应的鳟鱼 BAC，将对其进行部分测序以鉴定 LOH 缺失，允许确定与人类染色体的同线性 进一步鉴定潜在的候选抑癌基因 分析。该项目应提供对肾母细胞瘤的基本见解 使用实验上易于处理的低等脊椎动物模型进行致癌作用 系统。