Hepatic CYP enzymes and host-pathogen interactions

肝脏 CYP 酶和宿主-病原体相互作用

• 批准号: 8044836
• 负责人: Edward Thomas Morgan
• 金额: $ 36.97万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8044836
• 关键词: Address; Adverse effects; Affect; Alkane 1-monooxygenase; Bacteria; Bacterial Infections; Biochemical; Blood Circulation; CYP4A11 gene; Complement; Conditioned Culture Media; Cytochrome P450; Disease; Down-Regulation; Endotoxins; Enzymes; Escherichia coli; Escherichia coli Infections; Exhibits; Fatty Acids; Flavins; Food Poisoning; Funding; Gene Deletion; Gene Expression; Genetic; Health; Hepatic; Hepatocyte; Human; Immune response; Impairment; Individual; Infection; Inflammation; Inflammatory; Invaded; Kidney; Libraries; Liver; Measurement; Mixed Function Oxygenases; Modeling; Mus; Oxidative Stress; Pathology; Pharmaceutical Preparations; Play; Proteomics; Publishing; Regulation; Role; Serum; Severities; Signal Pathway; Signal Transduction; Stimulus; Therapeutic Effect; Tissues; Transgenic Mice; blood pressure regulation; cytokine; drug clearance; eicosanoid metabolism; enteropathogenic Escherichia coli; fatty acid metabolism; gastrointestinal; in vivo; lipid metabolism; liver metabolism; mutant; novel; oxidative damage; pathogen; response

DESCRIPTION (provided by applicant): Cytochrome P450 (P450) enzymes play crucial roles in the clearance of drugs from the circulation, and therefore changes in their activities can significantly influence the therapeutic and adverse effects of a drug on an individual. Infectious and inflammatory diseases cause the down-regulation of many P450s and other drug metabolizing enzymes in the liver, causing impairment of drug clearances. In mice infected with C. rodentium, a model of E. Coli food poisoning in humans, a number of P450 enzymes in the liver are moderately down-regulated. However, CYP4A enzymes are more profoundly affected, as is another drug metabolizing enzyme, i.e. flavin monooxygenase 3 (FMO3). Preliminary evidence suggests that one or more bacterially secreted factors regulate hepatic CYP4A expression in an endotoxin-independent manner. CYP4As have important roles in fatty acid metabolism, control of blood pressure and in oxidative stress. Therefore, it is important to understand how human CYP4A11 is regulated during infection, and to understand whether on not CYP4A regulation is a defensive or adaptive response of the host, or if the invading bacterium could be regulating CYP4As to its own advantage. To address these questions, the regulation of CYP4A11 will be studied in the livers and kidneys of CYP4A11 transgenic mice during C. rodentium infection. This will be complemented by studies in cultured human hepatocytes and in CYP4A11 transgenic mouse hepatocytes, on CYP4A11 regulation by inflammatory cytokines and bacterial products. Transcriptional and proteomic profiling will be used to assess the effects of infection on global gene expression in the liver and to elucidate signaling networks involved in CYP4A and FMO3 down-regulation by factors secreted by bacteria. To identify these factor(s) we will screen known mutants and, if necessary, an enteropathogenic E. coli transposon insertion library to identify mutants that fail to down-regulate these enzymes. This approach will be complemented by biochemical characterization and partial purification of the factor(s). To elucidate the role of CYP4A enzymes in the host response to C. rodentium infection, or as targets of the bacterium's strategy for propagation and survival, the effects will be studied of Cyp4A10-/- and Cyp4A14-/- gene deletion on the pathologies associated with C. rodentium infection. PUBLIC HEALTH RELEVANCE: This project will investigate the effects of a model of food poisoning on enzymes in the liver and kidneys called CYP4A enzymes, which are important in fat metabolism and also in oxidative damage to the tissues. The questions of whether the human enzyme levels are changed by infection in the same way as the mouse enzymes; how the liver responds to factors secreted by bacteria; and why these changes happen during infection, will be studied. This project will increase our understanding of how liver metabolism changes during infection or inflammation, and may discover new strategies to treat infectious and inflammatory diseases.

描述（由申请人提供）：细胞色素P450（P450）酶在清除循环中的药物中起着至关重要的作用，因此其活动的变化可以显着影响药物对个体的治疗和不良影响。传染病和炎症性疾病导致肝脏中许多P450和其他药物代谢酶的下调，从而导致药物清除障碍。在感染啮齿动物梭菌的小鼠中，人类大肠杆菌食物中毒的模型，肝脏中的许多P450酶被中度下调。然而，CYP4A酶受到更深远的影响，另一种药物代谢酶也受到了影响，即黄素单加氧酶3（FMO3）。初步证据表明，一个或多个细菌分泌的因素以内毒素独立的方式调节肝CYP4A的表达。 CYP4A在脂肪酸代谢，控制血压和氧化应激中具有重要作用。因此，重要的是要了解在感染过程中如何调节人类CYP4A11，并了解不进行CYP4A调节是宿主的防御或适应性反应，或者入侵细菌是否可以对CYP4AS对自己的优势进行调节。为了解决这些问题，将在啮齿动物念珠菌感染期间在CYP4A11转基因小鼠的肝脏和肾脏中研究CYP4A11的调节。这将通过对培养的人肝细胞和CYP4A11转基因小鼠肝细胞的研究，对CYP4A11通过炎症细胞因子和细菌产物调节的研究。转录和蛋白质组学分析将用于评估感染对肝脏全球基因表达的影响，并通过细菌分泌的因素来阐明参与CYP4A和FMO3涉及的信号网络。为了识别这些因素，我们将筛选已知的突变体，并在必要时筛选出肠病大肠杆菌转座子插入文库，以识别无法下调这些酶的突变体。这种方法将通过生化特征和因子的部分纯化来补充。 To elucidate the role of CYP4A enzymes in the host response to C. rodentium infection, or as targets of the bacterium's strategy for propagation and survival, the effects will be studied of Cyp4A10-/- and Cyp4A14-/- gene deletion on the pathologies associated with C. rodentium infection. 公共卫生相关性：该项目将研究食物中毒模型对称为CYP4A酶的肝脏和肾脏中的酶的影响，这些酶在脂肪代谢以及对组织的氧化损害中也很重要。感染是否与小鼠酶相同的方式改变了人类酶水平的问题；肝脏如何应对细菌分泌的因素；以及为什么在感染期间发生这些变化的原因。该项目将增加我们对肝脏代谢在感染或炎症过程中如何变化的理解，并可能发现治疗感染和炎症性疾病的新策略。