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（由适用提供）：电子烟（E-cig）被积极地作为烟草香烟的可行替代品。电子烟在美国和世界许多地区迅速获得接受，尤其是在儿童和青年中。但是，对电子烟/液体的化学分析表明，在各种电子烟产品中也发现了许多烟雾中存在的毒物和致癌物。尽管在电子烟产品中存在毒性和致癌物，但尚未在E-CIG使用者中研究暴露于这些污染物的生物学后果。这种创新的合作汇集了化学癌变，应用生物学和药物滥用和成瘾的临床心理学专家，他们将首次研究电子烟的生物学作用，这是由遗传，表观遗传学和转录症的早期标志物确定的，与癌症风险相关。我们的中心假设是“ E-cig气溶胶会对常规E-CIG使用者的口腔上皮细胞中的基因组，表观基因组和转录组造成不利影响”。 SA1：确定常规电子烟使用者口服上皮摄氏的E-CIG气溶胶的遗传毒性。使用经过验证的非侵入性刷牙技术，我们将从常规的E-cig使用者和非用户中收集口腔上皮细胞，然后量化整体基因组中DNA损伤的形成，并代表与癌症相关的基因（在核分辨率水平上）。 SA2：识别，分类和解释常规电子烟使用者口服上皮细胞中的全基因组DNA甲基化模式。与应用我们内部“ SEQ”基于“ SEQ”的甲基化检测测定法相比，我们将在E-CIG使用者中构建整个口服上皮细胞的DNA甲基甲基。 SA3：在常规电子烟使用者的口腔上皮细胞中建立全基因组的组蛋白修饰谱。与非用户相比，我们将使用chip-seq在e-cig用户的口腔上皮细胞中绘制活性和反射性组蛋白标记。 SA4：分析常规电子烟使用者的口腔上皮细胞中的全局基因表达谱。与非用户相比，我们将使用RNA-Seq在E-cig用户中构建口服上皮细胞的整个转录组。在我们所有的四个特定目标中，我们将递归分配的混合模型，以确定遗传和/或表观遗传学改变，转录调控和电子烟暴露指数（例如，使用频率，品牌和风味等）之间的关系。这将使我们能够识别具有功能上重要的遗传遗传学变化，这些变化特定于电子烟暴露并预测其健康后果。出于验证目的，我们将设计涉及siRNA敲低方法，敲除模型，脱甲基化剂或组蛋白脱乙酰基酶抑制剂的功能研究，以实验测试是否意外地将鉴定出的遗传性和/或表观遗传替代（S）意外地与口服癌变有关。最后，我们将将我们的数据与吸烟者基因组，表观基因组和转录组概况的公共领域数据库的数据集成在一起，以确定与吸烟相比使用电子烟的相对风险/益处。