Genetic epigenetic & transcripotomic effects of e-cig aerosol on oral epithelium

遗传表观遗传

• 批准号: 9117340
• 负责人: AHMAD BESARATINIA
• 金额: $ 40.11万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-03 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9117340
• 关键词: Aberrant DNA Methylation; Address; Aerosols; Affect; Biological; Biological Assay; Biological Products; Biology; Carcinogens; Cataloging; Catalogs; Cells; Cheek structure; Chemicals; Child; Cigarette; Clinical Psychology; Collaborations; Complement; CpG Islands; DNA; DNA Damage; DNA Footprint; DNA Methylation; Data; Databases; Detection; Development; Electronic cigarette; Epigenetic Process; Epithelial; Epithelial Cells; Evaluation; Exposure to; Flavoring; Frequencies; Gene Expression Profile; Genes; Genetic; Genetic Markers; Genome; Genomics; Health; Histone Deacetylase Inhibitor; Histones; Housing; Human; Interview; Investigation; Knock-out; Lead; Link; Liquid substance; Malignant Neoplasms; Maps; Marketing; Methylation; Minor; Modeling; Mutate; Mutation; Nicotine; Nucleotides; Oncogene Deregulation; Oncogenes; Oral; Oral cavity; Participant; Polymerase Chain Reaction; Proliferating; Public Domains; Public Health; Recovery; Recruitment Activity; Regulation; Relative Risks; Resolution; Risk; Site; Small Interfering RNA; Smoker; Substance Addiction; Substance abuse problem; TP53 gene; Techniques; Testing; The Cancer Genome Atlas; Time; Tobacco; Tobacco use; Transcriptional Regulation; Transferase; Tumor Suppressor Genes; United States; Validation; base; biomarker development; cancer risk; chemical carcinogenesis; chromatin immunoprecipitation; cigarette smoking; cigarette smoking; demographics; design; epigenome; epigenomics; genome-wide; genotoxicity; histone modification; indexing; innovation; interest; knock-down; methylation pattern; methylome; next generation sequencing; oral carcinogenesis; oral cavity epithelium; psychologic; public health relevance; reduce tobacco use; toxicant; transcriptome; transcriptome sequencing; transcriptomics; tumor; vapor

 DESCRIPTION (provided by applicant): Electronic cigarettes (e-cig) are aggressively marketed as viable alternatives to tobacco cigarettes. E-cig are rapidly gaining acceptance in the United States and many parts of the world, especially among children and youth. However, chemical analysis of e-cig vapor/liquid has shown that many toxicants and carcinogens present in cigarette smoke are also found in a wide range of e-cig products. Notwithstanding the presence of toxicants and carcinogens in e-cig products, the biological consequences of exposure to these contaminants have not been investigated in e-cig users. This innovative collaboration brings together experts in chemical carcinogenesis, applied biology, and clinical psychology of substance abuse and addiction who will investigate, for the first time, the biological effects of e-cig as determined by early markers of genetic, epigenetic, and transcriptomic effects linked to risk of cancer. Our central hypothesis is that "e-cig aerosol causes detrimental effects on the genome, epigenome, and transcriptome in oral epithelial cells of regular e-cig users". SA1: To determine the genotoxicity of e-cig aerosol in oral epithelial cels of regular e-cig users. Using a validated non-invasive brushing technique, we will collect oral epithelial cells from regular e-cig users and non-users, and subsequently quantify the formation of DNA damage in the genome overall and in representative cancer-related genes (at the nucleotide resolution level). SA2: To identify, catalogue, and interpret genome-wide DNA methylation patterns in oral epithelial cells of regular e-cig users. We will construct the whole DNA methylome of oral epithelial cells in e-cig users as compared to non-users applying our in-house `seq'-based methylation detection assay. SA3: To establish the genome-wide histone modification profiles in oral epithelial cells of regular e-cig users. We will use ChIP-seq to globally map active- and repressive histone marks in oral epithelial cells of e-cig users as compared to non-users. SA4: To analyze the global gene expression profile in oral epithelial cells of regular e-cig users. We will use RNA-seq to construct the whole transcriptome of oral epithelial cells in e-cig users as compared to non-users. In all our four Specific Aims, we will us recursively partitioned mixture models to determine relationships between genetic- and/or epigenetic alterations, transcriptional regulation, and e-cig exposure indices (e.g., frequency of use, brand and flavor, etc.). This will enable us to identify functionally important genetic- and/o epigenetic changes that are specific to e-cig exposure and predictive of its health consequences. For validation purposes, we will design functional studies involving siRNA knockdown approach, knockout models, demethylating agents, or histone deacetylase inhibitors to test experimentally whether any of the identified genetic- and/or epigenetic alteration(s) is causally linked to oral carcinogenesis. Lastly, we will integrate our data with those of public domain databases on smokers' genomic-, epigenomic-, and transcriptomic profiles to establish the relative risks/benefits of e-cig use compared to cigarette smoking.

 描述（由申请人提供）：电子香烟（e-cig）作为烟草香烟的可行替代品积极销售。电子烟在美国和世界许多地方，特别是在儿童和青年中迅速获得接受。然而，电子烟蒸气/液体的化学分析表明，香烟烟雾中存在的许多有毒物质和致癌物质也存在于各种电子烟产品中。尽管电子烟产品中存在有毒物质和致癌物质，但尚未对电子烟用户暴露于这些污染物的生物后果进行调查。这项创新的合作汇集了化学致癌作用，应用生物学和药物滥用和成瘾临床心理学的专家，他们将首次调查电子烟的生物学效应，这些效应是由与癌症风险相关的遗传，表观遗传和转录组学效应的早期标志物确定的。我们的中心假设是“电子烟气雾剂对常规电子烟使用者口腔上皮细胞的基因组、表观基因组和转录组造成有害影响”。SA 1：确定电子烟气雾剂对常规电子烟使用者口腔上皮细胞的遗传毒性。使用经过验证的非侵入性刷牙技术，我们将从常规电子烟使用者和非使用者中收集口腔上皮细胞，随后量化整个基因组和代表性癌症相关基因中DNA损伤的形成（在核苷酸分辨率水平上）。SA 2：识别、分类和解释常规电子烟使用者口腔上皮细胞中的全基因组DNA甲基化模式。我们将构建电子烟使用者口腔上皮细胞的整个DNA甲基化组，与非使用者相比，应用我们内部的基于“seq”的甲基化检测试验。SA 3：建立常规电子烟使用者口腔上皮细胞的全基因组蛋白修饰谱。我们将使用ChIP-seq在电子烟使用者与非使用者的口腔上皮细胞中全局映射活性和抑制性组蛋白标记。SA 4：分析常规电子烟使用者口腔上皮细胞中的整体基因表达谱。我们将使用RNA-seq构建电子烟使用者与非使用者口腔上皮细胞的全转录组。在我们的所有四个具体目标中，我们将使用递归分区混合模型来确定遗传和/或表观遗传改变、转录调控和电子烟暴露指数之间的关系（例如，使用频率、品牌和风味等）。这将使我们能够识别功能上重要的遗传和/或表观遗传变化，这些变化是特定于电子烟暴露并预测其健康后果的。出于验证目的，我们将设计涉及siRNA敲除方法、敲除模型、去甲基化剂或组蛋白去乙酰化酶抑制剂的功能性研究，以通过实验测试任何鉴定的遗传和/或表观遗传改变是否与口腔癌发生有因果关系。最后，我们将把我们的数据与公共领域数据库中关于吸烟者基因组、表观基因组和转录组特征的数据整合起来，以确定使用电子烟与吸烟相比的相对风险/益处。