DNA Methylation Based Biomarkers and Epigenetic Regulation in IBS

IBS 中基于 DNA 甲基化的生物标志物和表观遗传调控

• 批准号: 8968752
• 负责人: Lin Chang
• 金额: $ 23.1万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8968752
• 关键词: Abdominal Pain; Address; Adult; Adverse effects; Affect; Animals; Autonomic nervous system; Behavioral; Biological; Biological Assay; Biological Markers; Biopsy; Brain; Chronic; Chronic Cancer Pain; Chronic Disease; Colon; Complex; Constipation; DNA Methylation; Data; Development; Diagnosis; Diagnostic; Diarrhea; Disease; Environmental Risk Factor; Epigenetic Process; Event; Exclusion; Failure; Flare; Functional Gastrointestinal Disorders; Functional disorder; GSTM1 gene; Gastroenterology; Gastrointestinal Diseases; Gastrointestinal tract structure; Gene Expression Profiling; Genes; Genetic Markers; Glutathione S-Transferase; Goals; Habits; High Prevalence; Hormones; Human; Individual; Intestines; Irritable Bowel Syndrome; Knowledge; Lead; Link; Measures; Mental disorders; Methylation; Molecular; Monitor; Mucous Membrane; Neuraxis; Neuropeptides; Ontology; Oxidative Stress; Pain; Pathogenesis; Pathway interactions; Patients; Pattern; Peripheral Blood Mononuclear Cell; Permeability; Play; Population; Prevalence; Primary Health Care; Recurrence; Role; Sampling; Sensitivity and Specificity; Sensory Disorders; Site; Stress; Symptoms; Techniques; Testing; Time; Tissues; Tubulin; United States; Woman; accurate diagnosis; base; bisulfite sequencing; cell motility; chronic abdominal pain; cohort; drug discovery; effective therapy; epigenetic marker; epigenetic regulation; gastrointestinal; gene environment interaction; gene function; genome wide methylation; genome-wide; histone modification; human RBX1 protein; human SNCAIP protein; hypothalamic-pituitary-adrenal axis; insight; methylation biomarker; methylation pattern; motor disorder; next generation; novel diagnostics; peptide hormone; polymerization; public health relevance; stressor; treatment response

 DESCRIPTION (provided by applicant): Irritable bowel syndrome (IBS) is a chronic gastrointestinal disorder that affects up to 15% of adults, predominantly women, in the United States. Symptoms include chronic or recurrent abdominal pain associated with diarrhea, constipation or both. It is considered a stress-sensitive disorder that is associated with altered brain-gut interactions. Biomarkers that can reliably diagnose IBS or monitor treatment response are currently lacking. The current proposal is based on our preliminary data that epigenetic markers, namely DNA methylation, can distinguish IBS patients from healthy individuals. DNA methylation is a key epigenetic mechanism that governs vertebrate gene function. It has emerged as a leading mechanism linking gene- environment interactions to long-term behavioral development, particularly in complex disorders such as IBS. Our preliminary data on genome-wide DNA methylation for peripheral blood mononuclear cells (PBMCs) in a limited number of subjects (IBS: N=12, healthy controls: N=12) identified a set of epigenetic markers located on genes including SSPO, RNF39, GSTM1, GSTM5, TPPP and SNCAIP that can potentially distinguish IBS patients from healthy controls. Gene ontology analysis showed an enrichment of genes involved in neuropeptide pathways. We validated the differential methylation of CpG sites in these genes using bisulphite sequencing. However, these findings need to be replicated and validated in a larger cohort. Therefore, the studies proposed here are intended to identify robust epigenetic biomarkers for an accurate diagnosis of IBS and to gain critical insights into the pathogenesis of this disorder. Our aims are to: 1) Identify DNA methylation based biomarkers using genome-wide DNA methylation profiling in peripheral blood mononuclear cells (PBMCs) in IBS. 2) Study genome-wide methylation and expression differences in colon tissue of IBS patients and HCs. In Aim 1, we will conduct a genome-wide DNA methylation profiling in a larger, independent cohort of IBS patients (N=108) and controls (N=36) replicate the differential methylation of selected markers in a subset of samples using an affordable technique (MethyLight PCR) that can be employed for the routine diagnosis of IBS. In Aim 2, we propose genome-wide DNA methylation and gene expression profiling of banked colonic mucosal biopsies collected from IBS patients (N=108) and controls (N=36) to investigate epigenetically deregulated genes and associated pathways, which can give important insights on pathophysiology of IBS.

 描述（由申请人提供）：肠易激综合症 (IBS) 是一种慢性胃肠道疾病，影响美国多达 15% 的成年人，主要是女性。症状包括与腹泻、便秘或两者兼而有之的慢性或复发性腹痛。它被认为是一种与脑肠相互作用改变有关的压力敏感性疾病。目前缺乏能够可靠诊断 IBS 或监测治疗反应的生物标志物。目前的提议是基于我们的初步数据，即表观遗传标记，即 DNA 甲基化，可以区分 IBS 患者和健康个体。 DNA甲基化是控制脊椎动物基因功能的关键表观遗传机制。它已成为将基因-环境相互作用与长期行为发展联系起来的主要机制，特别是在IBS等复杂疾病中。我们对有限数量的受试者（IBS：N = 12，健康对照：N = 12）外周血单核细胞（PBMC）全基因组 DNA 甲基化的初步数据确定了一组位于 SSPO、RNF39、GSTM1、GSTM5、TPPP 和 SNCAIP 等基因上的表观遗传标记，可以将 IBS 患者与健康对照区分开来。基因本体分析显示参与神经肽途径的基因丰富。我们使用亚硫酸氢盐测序验证了这些基因中 CpG 位点的差异甲基化。然而，这些发现需要在更大的队列中进行复制和验证。因此，本文提出的研究旨在确定强大的表观遗传生物标志物，以准确诊断 IBS 并获得对该疾病发病机制的重要见解。我们的目标是：1) 使用 IBS 外周血单核细胞 (PBMC) 的全基因组 DNA 甲基化分析来识别基于 DNA 甲基化的生物标志物。 2) 研究IBS患者和HC结肠组织的全基因组甲基化和表达差异。在目标 1 中，我们将在更大的独立 IBS 患者组 (N=108) 和对照组 (N=36) 中进行全基因组 DNA 甲基化分析，使用可用于 IBS 常规诊断的经济实惠的技术 (MethyLight PCR) 在一部分样本中复制选定标记物的差异甲基化。在目标 2 中，我们提出对从 IBS 患者 (N=108) 和对照 (N=36) 收集的结肠粘膜活检样本进行全基因组 DNA 甲基化和基因表达谱分析，以研究表观遗传失调基因和相关通路，这可以为 IBS 的病理生理学提供重要见解。