Molecular Analysis of Alcohol Effects on Human Oral Epithelium Cells' Epigenome

酒精对人口腔上皮细胞表观基因组影响的分子分析

• 批准号: 8256369
• 负责人: Alison Urvalek
• 金额: $ 4.84万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8256369
• 关键词: Acetaldehyde; Address; Affect; Aftercare; Alcohol abuse; Alcoholism; Alcohols; Benzo(a)pyrene; Carcinogen exposure; Carcinogens; Cells; Cessation of life; Code; Collaborations; Colon Carcinoma; DNA; DNA Methylation; DNA Methyltransferase Inhibitor; DNA Sequence; Detection; Diagnosis; Disease; Early Diagnosis; Epigenetic Process; Epithelial Cells; Ethanol; Future; Gene Expression; Gene Silencing; Genes; Goals; Head and Neck Cancer; Head and neck structure; Histones; Human; Institutes; Intercistronic Region; Lead; Malignant Neoplasms; Malignant neoplasm of liver; Malignant neoplasm of lung; Measures; Mediating; Mission; Molecular; Molecular Analysis; National Institute of Dental and Craniofacial Research; Oral; Pharmaceutical Preparations; Post-Translational Protein Processing; Prevention; Public Health; RNA; RNA Sequences; Research; Risk Factors; Role; Survival Rate; Techniques; Testing; Tobacco; Tobacco use; Tobacco-Associated Carcinogen; Tretinoin; Tumor Initiators; Tumor Suppressor Genes; Vitamin A; alcohol abuse therapy; alcohol effect; alcohol exposure; bisulfite; cancer initiation; cancer risk; carcinogenesis; chromatin immunoprecipitation; effective therapy; epigenomics; genome-wide; histone modification; improved; innovation; keratinocyte; mRNA Expression; malignant breast neoplasm; malignant mouth neoplasm; oral carcinogenesis; oral cavity epithelium; prevent; prognostic; promoter; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): In addition to tobacco, alcohol is the major risk factor for head and neck cancer. Some of the initial steps of head and neck carcinogenesis are believed to be epigenetic changes, which are defined as changes in gene expression that are not a result of alterations in the underlying DNA coding sequence. Alcohol and its metabolite acetaldehyde (AcH) promote epigenetic changes in breast, liver, and colon cancers, but whether or not genome-wide epigenetic changes are induced by alcohol in oral epithelial cells is unknown. The long term goal of these studies is to understand the molecular mechanisms by which alcohol promotes head and neck tumorigenesis. The objective of this application is to understand whether alcohol regulates genome-wide epigenetic changes, such as DNA methylation and histone modifications in intergenic regions and at the promoters of genes whose expression is changed during human oral carcinogenesis. We will also determine whether retinoic acid (RA) and the DNA methyltransferase inhibitor 5-Aza-2'-deoxycitidine (5-Aza) can reverse epigenetic changes induced by the tobacco carcinogen benzo[a]pyrene (B[a]P) and alcohol. Our central hypothesis is that alcohol and carcinogens found in tobacco will induce genome-wide aberrant epigenetic changes that will alter gene expression of normal human oral epithelial cells, and that retinoic acid and DNA methyltransferase inhibitors will reverse alcohol and carcinogen associated epigenetic changes. Two specific aims will test this hypothesis: Aim 1: To determine if alcohol regulates epigenetic changes and if these changes correlate with changes in gene expression in human oral epithelial cells. Genome-wide sequencing of ethanol treated immortalized OKF6-TERT1 human oral epithelial cells will be performed with the assistance of our institute's Epigenetic Core. This core has expertise in our proposed approaches which include: Reduced Representation Bisulfite Sequencing (RRBS), Chromatin Immunoprecipitation sequencing (ChIP-seq), and RNA-sequencing (RNA-seq) techniques, to determine genome-wide changes in DNA methylation, histone post-translational modifications, and changes in mRNA expression after alcohol treatment, respectively. Aim 2: To determine the role of RA and DNA methyltransferase inhibitors in regulating the epigenome of human oral epithelial cells after alcohol and carcinogen exposure. Cells treated with B[a]P and ethanol will be subjected to RRBS, ChIP-seq, and RNA-seq to determine epigenetic changes and changes in gene expression. We will also examine whether B[a]P and ethanol induced epigenetic changes can be reversed by RA and 5-Aza treatment. This research is innovative because it will use the most cutting edge genome-wide sequencing techniques used to study epigenetic changes. The proposed research is significant as it will fulfill the mission of the National Institute of Dental and Craniofacial Research (NIDCR) and the National Institute of Alcohol Abuse and Alcoholism (NIAAA) by identifying some of the initial steps during carcinogen and alcohol mediated oral carcinogenesis, which will lead to better preventative, prognostic, and treatment approaches for this disease. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health because it will provide a mechanism for how alcohol can promote head and neck cancers. These studies will also allow us to identify changes that occur in cells during the initial steps of cancer initiation and progression, and to determine whether certain drug treatments can reverse the changes induced by alcohol. As a result, these studies will allow for better prevention, diagnosis, and treatment of head and neck cancers.

描述（由申请人提供）：除了烟草之外，酒精也是头颈癌的主要危险因素。头颈癌发生的一些初始步骤被认为是表观遗传变化，表观遗传变化被定义为基因表达的变化，而不是潜在 DNA 编码序列改变的结果。酒精及其代谢物乙醛 (AcH) 会促进乳腺癌、肝癌和结肠癌的表观遗传变化，但酒精是否会在口腔上皮细胞中诱导全基因组表观遗传变化尚不清楚。这些研究的长期目标是了解酒精促进头颈部肿瘤发生的分子机制。本申请的目的是了解酒精是否调节全基因组表观遗传变化，例如基因间区域和人类口腔癌发生过程中表达发生变化的基因启动子中的 DNA 甲基化和组蛋白修饰。我们还将确定视黄酸 (RA) 和 DNA 甲基转移酶抑制剂 5-Aza-2'-脱氧胞苷 (5-Aza) 是否可以逆转烟草致癌物苯并[a]芘 (B[a]P) 和酒精引起的表观遗传变化。我们的中心假设是，烟草中发现的酒精和致癌物会诱导全基因组异常表观遗传变化，从而改变正常人类口腔上皮细胞的基因表达，而视黄酸和 DNA 甲基转移酶抑制剂将逆转酒精和致癌物相关的表观遗传变化。有两个具体目标将检验这一假设： 目标 1：确定酒精是否调节表观遗传变化，以及这些变化是否与人类口腔上皮细胞基因表达的变化相关。将在我们研究所表观遗传核心的协助下对乙醇处理的永生化 OKF6-TERT1 人类口腔上皮细胞进行全基因组测序。该核心在我们提出的方法方面拥有专业知识，包括：简化代表性亚硫酸氢盐测序 (RRBS)、染色质免疫沉淀测序 (ChIP-seq) 和 RNA 测序 (RNA-seq) 技术，分别确定 DNA 甲基化、组蛋白翻译后修饰和酒精处理后 mRNA 表达变化的全基因组变化。目标 2：确定 RA 和 DNA 甲基转移酶抑制剂在酒精和致癌物暴露后调节人类口腔上皮细胞表观基因组的作用。用 B[a]P 和乙醇处理的细胞将接受 RRBS、ChIP-seq 和 RNA-seq 以确定表观遗传变化和基因表达的变化。我们还将研究 RA 和 5-Aza 治疗是否可以逆转 B[a]P 和乙醇诱导的表观遗传变化。这项研究具有创新性，因为它将使用最前沿的全基因组测序技术来研究表观遗传变化。拟议的研究意义重大，因为它将通过确定致癌物和酒精介导的口腔癌发生过程中的一些初始步骤来履行国家牙科和颅面研究所 (NIDCR) 和国家酒精滥用和酒精中毒研究所 (NIAAA) 的使命，这将导致针对这种疾病的更好的预防、预后和治疗方法。 公共健康相关性：拟议的研究与公共健康相关，因为它将提供酒精如何促进头颈癌的机制。这些研究还将使我们能够识别在癌症发生和发展的最初阶段细胞中发生的变化，并确定某些药物治疗是否可以逆转酒精引起的变化。因此，这些研究将有助于更好地预防、诊断和治疗头颈癌。