Genomic profiling of yeast resistance to heterocylic aromatic amines

酵母对杂环芳香胺的抗性的基因组分析

基本信息

批准号：
8626657
负责人：
MICHAEL Thomas FASULLO
金额：
$ 30.8万
依托单位：
STATE UNIVERSITY OF NEW YORK AT ALBANY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-12-09 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8626657
关键词：
2-Amino-1-Methyl-6-Phenylimidazo[4,5-b]pyridine Adult Aflatoxin B1 Alleles Aromatic Amines Bar Codes Base Excision Repairs Benzo(a)pyrene Bioinformatics Biological Assay CYP1A1 gene CYP1A2 gene CYP1B1 gene CYP2E1 gene CYP3A4 gene Cancer Biology Cancer Etiology Carcinogens Cell Death Cell Separation Cell physiology Cells Cellular biology Cessation of life Chemical Agents Chromosome Mapping Collection Colon Colon Carcinoma Colorectal Cancer Computational Technique Computer Analysis Cytochrome P450 DNA DNA Adducts DNA Damage DNA Repair Diet Diploidy Environmental Carcinogens Enzymes Epidemiologic Studies Family Gene Deletion Genes Genetic Genetic Polymorphism Genomics Haploidy Health Personnel High-Throughput DNA Sequencing High-Throughput Nucleotide Sequencing Housekeeping Gene Human Individual Libraries Link Liver Malignant Neoplasms Measures Meat Metabolic Activation Metabolism Methods Microscopy Mismatch Repair Molecular Mutagens Nucleotide Excision Repair Ontology Oxidoreductase Pathway interactions Phenotype Plasmids Population Quinoxalines Research Personnel Resistance Risk Saccharomyces cerevisiae Saccharomycetales Sample Size Sampling Smoking Students System Systems Biology Toxic effect Toxicology Training Transferase United States Yeasts adduct base cancer cell cell growth cooking deletion library design fetal gene function genotoxicity graduate student heterocyclic aromatic amines high throughput analysis killings lifestyle factors man mutant public health relevance quinoline recombinational repair red meat consumption research study resistant strain response tool undergraduate student yeast genome

项目摘要

Abstract/Summary Colorectal cancer is the second leading cause of cancer deaths in the United States. Fewer than 5% of colon cancer cases are linked to known high-penetrant genetic factors, while lifestyle factors include a diet high in red meat consumption. Carcinogens found in cooked red meat include heterocyclic aromatic amines (HAAs), which become potent DNA damaging agents after activation by P450 enzymes and N-acetyl transferase 2 (NAT2). However, it is unknown which individuals are most susceptible to HAAs and epidemiological studies often lack significance because of small sampling sizes. Since many DNA metabolism and housekeeping genes are conserved from yeast to man, high-throughput analysis of Saccharomyces cerevisiae (budding yeast) genes that confer resistance to carcinogens have identified human genes that confer resistance to environmental carcinogens. Genomic phenotyping using the ~5,000 yeast single-gene deletion haploid and diploid strains have been highly successful in determining genes that confer resistance to chemical agents. We previously were successful in introducing human CYP1A2 and CYP1A1 into yeast and activating a variety of carcinogens, including aflatoxin B1 (AFB1), benzo[a]pyrene dihydrodiol (BaP-DHD), and HAAs. The aim of this project is to determine which yeast genes are required for resistance to the potent HAA carcinogens. We will introduce plasmids that express human CYP1A2 and NAT2 into the ~5,000 diploid homozygous single-deletion strains. In the first aim, we will profile the yeast genome using the diploid single deletion strains for resistance to HAAs. Genes that confer resistance will be identified by high-throughput sensitive assays to measure cell growth and by molecular bar codes using high throughput sequencing. Considering that CYP1A2 is mostly expressed in the liver and not the colon, in the second aim, we will identify which colon-associated CYPs can metabolically activate HAAs. The information resulting from this project will aid health care providers in identifying individuals most at risk for colon cancer due to dietary carcinogens. The project will be a valuable training tool for graduate and undergraduate students in systems and cell biology.

摘要/总结结直肠癌是美国癌症死亡的第二大原因。小于结肠的 5% 癌症病例与已知的高渗透遗传因素有关，而生活方式因素包括高渗透性饮食红肉消费。煮熟的红肉中发现的致癌物质包括杂环芳香胺（HAAs）、被 P450 酶和 N-乙酰转移酶 2 激活后成为有效的 DNA 损伤剂（NAT2）。然而，尚不清楚哪些人最容易受到 HAA 的影响和流行病学研究由于样本量较小，通常缺乏显着性。由于许多 DNA 代谢和内务管理基因从酵母到人类都是保守的，酿酒酵母（出芽）的高通量分析酵母）赋予致癌物抗性的基因已经鉴定出赋予致癌物抗性的人类基因环境致癌物。使用约 5,000 个酵母单基因缺失单倍体和二倍体菌株在确定赋予化学试剂抗性的基因方面非常成功。我们此前已成功将人CYP1A2和CYP1A1引入酵母并激活多种致癌物，包括黄曲霉毒素 B1 (AFB1)、苯并[a]芘二氢二醇 (BaP-DHD) 和 HAAs。此举的目的该项目的目的是确定哪些酵母基因是抵抗强效HAA致癌物所必需的。我们将将表达人 CYP1A2 和 NAT2 的质粒引入约 5,000 个二倍体纯合单缺失中菌株。第一个目标是，我们将使用二倍体单缺失菌株来分析酵母基因组的抗性到HAA。赋予抗性的基因将通过高通量敏感测定来鉴定以测量细胞生长和使用高通量测序的分子条形码。考虑到 CYP1A2 主要是在肝脏而不是结肠中表达，在第二个目标中，我们将确定哪些结肠相关 CYP 可以代谢激活HAA。该项目产生的信息将帮助医疗保健提供者识别因膳食致癌物而最有患结肠癌风险的个体。该项目将是一个有价值的项目系统和细胞生物学研究生和本科生的培训工具。