Effect of ethanol on retinoid metabolism and signaling in zebrafish embryos

乙醇对斑马鱼胚胎中类维生素A代谢和信号传导的影响

• 批准号: 7142426
• 负责人: WILLIAM F BOSRON
• 金额: $ 17.99万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7142426
• 关键词: biological signal transduction; bone development; craniofacial; developmental genetics; developmental neurobiology; embryo /fetus toxicology; ethanol; fetal alcohol syndrome; fluorescent dye /probe; gene expression; genetic regulation; in situ hybridization; labyrinth; morphometry; nonmammalian vertebrate embryology; phenotype; recombinant proteins; retinoate; retinoid binding proteins; teratogens; vitamin metabolism; vitamin receptor; zebrafish

DESCRIPTION (provided by applicant): This is an R21 exploratory/development grant application to develop methods to quantify the content of alcohol metabolizing enzymes and variants (initially alcohol dehydrogenase and eventually aldehyde dehydrogenase and microsomal ethanol oxidizing CYP isoenzymes) in human tissues (liver and esophagus). We believe that the application fits the R21 guidelines because we will use newly developed mass spectrometry proteomics methods to address the specific aims. The application is also in response to PA-05-074, Mechanisms of Alcohol-Induced Tissue Injury, because it provides important protein expression information to support the investigation of alcohol metabolizing enzyme genotypes that confer susceptibility or resistance to ethanol-induced cell/tissue damage. Our central hypothesis is that the expression/content of alcohol metabolizing enzymes and variants determine the pharmacokinetics and cellular toxicology of ethanol and acetaldehyde in tissues. In Aim #1, we will develop proteomics methods to quantify the expression of seven alcohol dehydrogenase (ADH) isoenzymes in tissues by triple quadrupole linear ion trap LC/MS/MS and multiple reactions monitoring analysis with appropriate peptide/protein internal standards. We will assess the feasibility of developing methods to look at the whole set of ADH, ALDH and microsomal ethanol oxidizing CYP isoenzymes involved in alcohol metabolism simultaneously. In Aim #2, we will collect and genotype human liver samples and matched normal esophagus and squamous cell carcinoma tissue samples from a local population in Indianapolis for the pilot proteomics study. The tissue samples will be genotyped at ADH and other alcohol metabolizing enzyme loci. In Aim #3, we will perform a preliminary proteomics study to look at the variability of expression/content of the seven ADH isoenzymes in the liver and esophagus tissue samples using the proteomics methods developed in Aim #1. We will identify whether there is a correlation between ADH polymorphisms and protein expression in tissues. We will estimate the individual contribution of the seven ADHs to alcohol metabolic capacity in tissue samples and the contribution of polymorphism to differences in alcohol metabolic capacity. We will assess the feasibility of performing targeted proteomics studies with tissues from other populations that express different SNPs in alcohol metabolizing enzymes.

描述（由申请人提供）：这是R21探索/开发赠款应用程序，用于开发量化酒精代谢酶和变体含量的方法（最初是酒精脱氢酶以及最终的醛脱氢酶和微体乙醇氧化乙醇氧化酶（Liver和Esophagus和Esophagus））。我们认为该应用程序适合R21指南，因为我们将使用新开发的质谱蛋白质组学方法来解决特定目标。该应用也是对PA-05-074的响应，即酒精诱导的组织损伤的机制，因为它提供了重要的蛋白质表达信息，以支持对酒精代谢酶基因型的研究，从而赋予乙醇诱导的细胞/组织损伤的易感性或抗性。我们的中心假设是，酒精代谢酶和变体的表达/含量决定了组织中乙醇和乙醛的药代动力学和细胞毒理学。在AIM＃1中，我们将开发蛋白质组学方法来量化通过三倍四极线离子陷阱在组织中七个酒精脱氢酶（ADH）同工酶的表达 LC/MS/MS和具有适当肽/蛋白质内部标准标准的多个反应监测分析。我们将评估开发方法来研究ADH，ALDH和微粒体乙醇氧化的CYP同工酶的整个方法的可行性。在AIM＃2中，我们将收集和基因型人肝样品，并与印第安纳波利斯当地人群的正常食道和鳞状细胞癌组织样品进行匹配，以进行PILOT蛋白质组学研究。组织样品将在ADH和其他酒精代谢酶位点进行基因分型。在AIM＃3中，我们将使用AIM＃1中开发的蛋白质组学方法进行初步的蛋白质组学研究，以研究肝脏和食道组织样品中七个ADH同工酶的表达/含量的可变性。我们将确定ADH多态性与组织中蛋白质表达之间是否存在相关性。我们将估计七种ADH对组织样品中酒精代谢能力的个人贡献以及多态性对酒精代谢能力差异的贡献。我们将评估通过在酒精代谢酶中表达不同SNP的其他人群的组织进行靶向蛋白质组学研究的可行性。