Gene Expression in Barrett's Esophagus

巴雷特食管中的基因表达

• 批准号: 6927350
• 负责人: Hashem B El-Serag
• 金额: $ 15万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6927350
• 关键词: Barretts esophagus; biopsy; cadherins; clinical research; endoscopy; esophagus neoplasm; gastrointestinal epithelium; gastrointestinal system; gene expression; genetic susceptibility; histopathology; human subject; laser capture microdissection; microarray technology; neoplasm /cancer genetics; pathologic process; polymerase chain reaction; preneoplastic state; reflux esophagitis

DESCRIPTION (provided by the applicant): Background: Barrett's esophagus (BE) is the precursor lesion for esophageal adenocarcinoma. While there is information on isolated somatic changes that occur during the progression of BE to dysplasia or cancer, the global changes in somatic gene expression underlying the transformation of normal esophageal mucosa to BE remain unknown. Hypothesis: BE is associated with altered gene expression patterns. Comparative gene expression studies using microarray analyses would identify BE-specific genes that provide insight into the molecular basis of BE. Our preliminary data indicate an overactive Wnt/beta-catenin pathway in BE. Specific Aim #1: To identify the specific gene expression pattern for Barrett's esophagus. a) Recruit individuals from three well-defined phenotypically distinct cohorts: patients with BE, patients with GERD and no BE, and volunteers with no GERD or BE. b) Obtain endoscopically and histologically well-defined tissue classes from cohorts identified in SA#1.a.; these tissue classes are BE, normal esophageal, gastric, duodenal, and colonic epithelia. c) Use laser microdissection to capture glandular tissue. d) Use microarray analysis to compare the global gene expression of BE to several control tissues obtained from the three cohorts (SA# 1.a and 1.b). Specific Aim #2: To identify novel gene(s) and/or pathways in the pathogenesis of Barrett's esophagus. a) Apply the gene expression data to metabolic and/or physiological pathways with sample size calculation geered toward the Wnt/beta-catenin pathway. b) Use quantitative RT-PCR to confirm the altered expression of BE-specific genes. Methods: We will obtain endoscopic biopsies from esophageal, gastric, duodenal, and colonic tissue from 30 patients (10 with BE, 10 with GERD w/o BE, and 10 with no GERD or BE). We will conduct microarray assays using RNA extracted from these specimens on Human Genome Affymetrix (HG- 133U) Set GeneChip(r). The data will be analyzed to identify downregulated and upregulated genes that are unique to BE. Pathway surveys will be conducted with confirmation of these genes using RT-Q-PCR. We present preliminary data on microarray and RT-PCRanalyses on 5 patients with BE that suggest an overactive Wnt pathway: we have also assembled the essential components of a successful study using microarrays.

描述（由申请人提供）： 背景：巴雷特食管（BE）是食管腺癌的先兆病变。虽然有关于BE进展为不典型增生或癌症期间发生的孤立体细胞变化的信息，但正常食管粘膜转化为BE过程中体细胞基因表达的整体变化仍然未知。假设：BE 与基因表达模式的改变有关。使用微阵列分析的比较基因表达研究将识别 BE 特异性基因，从而深入了解 BE 的分子基础。我们的初步数据表明 BE 中 Wnt/β-catenin 通路过度活跃。具体目标#1：确定巴雷特食管的特定基因表达模式。 a) 从三个明确定义的表型不同的队列中招募个体：BE 患者、GERD 但无 BE 患者以及无 GERD 或 BE 的志愿者。 b) 从 SA#1.a. 中确定的群组中获得内窥镜和组织学上明确的组织类别；这些组织类别是 BE、正常食管、胃、十二指肠和结肠上皮。 c) 使用激光显微切割捕获腺体组织。 d) 使用微阵列分析将 BE 的整体基因表达与从三个队列（SA# 1.a 和 1.b）获得的几个对照组织进行比较。具体目标#2：确定巴雷特食管发病机制中的新基因和/或途径。 a) 将基因表达数据应用于代谢和/或生理途径，并针对 Wnt/β-连环蛋白途径进行样本量计算。 b) 使用定量 RT-PCR 确认 BE 特异性基因表达的改变。方法：我们将从 30 名患者的食管、胃、十二指肠和结肠组织中获取内窥镜活检（10 名患有 BE，10 名患有 GERD 不伴有 BE，10 名没有 GERD 或 BE）。我们将使用从这些样本中提取的 RNA 在人类基因组 Affymetrix (HG-133U) Set GeneChip(r) 上进行微阵列分析。将对数据进行分析，以确定 BE 特有的下调和上调基因。将使用 RT-Q-PCR 确认这些基因并进行通路调查。我们提供了 5 名 BE 患者的微阵列和 RT-PCR 分析的初步数据，表明 Wnt 通路过度活跃：我们还使用微阵列组装了一项成功研究的基本组成部分。