Regulation of PTEN in chronic alcohol mediated liver disease

PTEN 在慢性酒精介导的肝病中的调节

基本信息

批准号：
8262183
负责人：
Colin Shearn
金额：
$ 6.2万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-01 至 2013-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8262183
关键词：
1-Phosphatidylinositol 3-Kinase 4 hydroxynonenal Active Sites Address Affect Alcoholic Hepatitis Alcoholic Liver Cirrhosis Alcoholic Liver Diseases Alcohols Aldehydes Amino Acids Chronic Cirrhosis Complex Cysteine DNA Data Development Diagnosis Enzymes Ethanol Ethanol toxicity Fatty Liver Hepatocyte Hydrogen Peroxide Immunohistochemistry In Vitro Lead Lipid Binding Lipid Peroxidation Lipids Liquid substance Liver Liver diseases Mass Spectrum Analysis Mediating Metabolism Modeling Modification Morbidity - disease rate Mus Oxidative Stress PDPK1 gene PTEN gene Pathway interactions Phosphatidylinositols Phosphoric Monoester Hydrolases Physiological Positioning Attribute Property Proteins Proto-Oncogene Proteins c-akt Publications Rattus Recombinant Proteins Recombinants Regulation Reporting Research Proposals Role Schiff Bases Signal Pathway Signal Transduction Signaling Protein Site Survival Rate Symptoms System Testing Thioredoxin Time Tissues Tumor Suppressor Proteins United States Western Blotting Work adduct alcohol abuse therapy alcohol exposure cell growth chronic alcohol ingestion in vitro Model in vivo inorganic phosphate insight mRNA Expression membrane activity mortality mouse model mutant pi bond research study statistics

项目摘要

The objective of this proposal is to determine the involvement of the lipid phosphatase PTEN in the formation of steatosis and liver damage during alcoholic liver disease (ALD). Although there are publications concerning the role of PTEN in ALD, these reports are using a static time point in a fluid model and do not address the possibility of both PTEN inactivation via aldehyde modification and changes in PTEN regulation/expression over a period of alcohol exposure. Our working hypothesis states the reactive aldehyde 4-HNE, produced in the liver of chronic ethanol treated mice covalently adducts PTEN thereby reducing enzymatic activity leading to disregulation in PTEN downstream signaling. 4-HNE has already been implicated in the inactivation of proteins during chronic ethanol exposure in the liver. It is also hypothesized that increased PTEN expression leads to changes in downstream pathways ultimately leading to increased steatosis seen in ALD. In order to fulfill the objectives of this research proposal, experiments will be performed using mass spectrometry to identify the sites of 4-HNE modification on PTEN, the ability of these adducts to inhibit enzymatic activity, membrane association and Trx1 association. In addition, using western blotting, immunohistochemistry and mRNA expression, the effects of variable PTEN expression on downstream signaling pathways will be examined following 9-weeks of chronic ethanol exposure.

该提案的目的是确定脂质磷酸酶 PTEN 在酒精性肝病 (ALD) 期间脂肪变性和肝损伤形成中的参与情况。尽管有一些出版物涉及 PTEN 在 ALD 中的作用，但这些报告在流体模型中使用静态时间点，并没有解决通过醛修饰导致 PTEN 失活以及在酒精暴露一段时间内 PTEN 调节/表达发生变化的可能性。我们的工作假设指出，长期乙醇处理的小鼠肝脏中产生的反应性醛 4-HNE 与 PTEN 共价加合物，从而降低酶活性，导致 PTEN 下游信号传导失调。 4-HNE 已被证实与肝脏长期接触乙醇期间蛋白质失活有关。还假设 PTEN 表达增加会导致下游通路发生变化，最终导致 ALD 中脂肪变性增加。为了实现本研究计划的目标，将使用质谱法进行实验来鉴定 PTEN 上 4-HNE 修饰的位点、这些加合物抑制酶活性、膜关联和 Trx1 关联的能力。此外，将使用蛋白质印迹、免疫组织化学和 mRNA 表达，在慢性乙醇暴露 9 周后检查可变 PTEN 表达对下游信号通路的影响。