Expression and Post-translational Modification of Cytochrome P450 4A11 Variants in Modulation of Blood Pressure Through Regulation of 20-Hydroxyeicosatetraenoic Acid (20-HETE) Production

细胞色素 P450 4A11 变体的表达和翻译后修饰通过调节 20-羟基二十碳四烯酸 (20-HETE) 的产生来调节血压

• 批准号: 9393139
• 负责人: Matthew Edward Albertolle
• 金额: $ 2.86万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2019-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9393139
• 关键词: Affect; Age; Agonist; Alkane 1-monooxygenase; Alleles; Antihypertensive Agents; Arachidonic Acids; Autacoids; Biochemical; Biological Assay; Blood Pressure; CYP4A11 gene; Complex; Coronary heart disease; Cytochrome P450; Data; Diuretics; Economic Burden; Eicosanoids; Environment; Enzymes; Family; Genetic Variation; Genotype; Goals; Healthcare Systems; Homologous Gene; Human; Hydrogen Peroxide; Hydroxyeicosatetraenoic Acids; Hydroxylation; Hypertension; In Vitro; Individual; Isotope Labeling; Kidney; Lead; Limb structure; Liver; Mass Spectrum Analysis; Measurement; Measures; Microsomes; Mixed Function Oxygenases; Modeling; Mus; Natriuresis; Nature; Oxidation-Reduction; Oxides; Pathway interactions; Patients; Persons; Phenotype; Play; Polyunsaturated Fatty Acids; Post-Translational Protein Processing; Post-Translational Regulation; Predisposition; Production; Proteins; Reactive Oxygen Species; Regulation; Reporting; Resistance; Risk; Rodent; Role; Sampling; Scheme; Signal Transduction; Site; Sodium Chloride; Stroke; Sulfenic Acids; Sulfhydryl Compounds; System; Testing; Thick; Transgenic Mice; Transgenic Organisms; Variant; Vasoconstrictor Agents; blood pressure reduction; blood pressure regulation; experimental study; genetic variant; mouse model; novel therapeutics; oxidation; response; saluretic; tool; treatment response

ABSTRACT Hypertension is a complex condition that affects 40% of persons over the age of 25 years. Reduction in blood pressure has been associated with a significant decrease in both coronary heart disease and stroke. Our long term goal is to develop novel therapeutic strategies for hypertension in the context of the cytochrome P450 system. Cytochrome P450 (CYP) 4A11 is an -hydroxylase that catalyzes the oxidation of arachidonic acid (AA) to 20-hydroxyeicosatetraenoic acid (20-HETE). 20-HETE can be pro-hypertensive by constricting microvasculature or anti-hypertensive by inhibiting salt reabsorption in the kidney. Thus, 20-HETE and CYP4A11 have important roles in blood pressure regulation. In humans, a genetic variant of CYP4A11 (rs1126742 T8590C allele) encodes for a protein (CYP4A11-F434S) with reduced -hydroxylation activity that has been associated with high blood pressure and resistance to the anti-hypertensive effects of some diuretics. This apparent deregulation, in conjunction with decreased 20-HETE activity in the F434S variant, suggests that other unknown factors may contribute to the pro- and anti-hypertensive effects of CYP4A11. The central hypothesis is that expression and/or posttranslational modification differentially effects genetic variants of CYP4A11 with regard to 20-HETE synthesis and, ultimately, blood pressure control. CYP4A11 regulation and/or enzymatic activity will be tested for effects on 20-HETE levels and for influences by reactive oxygen species (ROS). A combination of biochemical, analytical, and enzymological tools will be used to further elucidate the differences between the CYP4A11 Phe-434 and Ser-434 variants in substrate selection, activity, and protein level. First, the use of chemoselective probes will determine the role of redox status in the posttranslational regulation in CYP4A11. Next, the relationship between CYP4A11 variant protein concentration and arachidonic acid metabolizing activity will be investigated in human microsomes and a transgenic mouse model. These experiments aim to further elucidate the role CYP4A11 variants play in hypertension. 20-HETE is an important component in hypertension. Genetic variations that alter its production cause significant changes in hypertension susceptibility and therapeutic response. Understanding the mechanisms related to the phenotypic variation in CYP4A11 will help to gain a more in-depth understanding of hypertension in different individuals and reshape how these specific persons are treated for blood pressure reduction.

抽象的 高血压是一种复杂的疾病，影响了25岁以上40％的人。减少血液 压力与冠状动脉疾病和中风都显着降低有关。我们的漫长 术语目标是在细胞色素P450的背景下制定新型的高血压治疗策略 系统。细胞色素P450（CYP）4A11是羟化酶，可催化蛛网膜酸的氧化 （AA）至20-羟基二乙烯烯酸（20-HETE）。 20-Hete可以通过限制来促成贝雷重 通过抑制肾脏的盐吸收，微举行或抗高血压。那，20-hete和 CYP4A11在血压调节中具有重要作用。在人类中，CYP4A11的遗传变异 （RS1126742 T8590C等位基因）编码蛋白质（CYP4A11-F434S），其降低了-羟基化活性 与某些利尿剂的抗高血压作用有关，与高血压和抗性有关。 这种明显的放松管制与F434S变体中20-HETE活性的降低相结合，表明 其他未知因素可能有助于CYP4A11的促和抗高血压作用。中央 假设是表达和/或翻译后修饰对遗传变异的影响不同 CYP4A11关于20-HETE合成，最终是血压控制。 CYP4A11法规 和/或酶促活性将测试对20-HETE水平的影响以及活性氧的影响 物种（ROS）。生化，分析和酶学工具的结合将用于进一步 阐明CYP4A11 PHE-434和Ser-434变体之间的差异 和蛋白质水平。首先，化学选择性问题的使用将决定氧化还原状态在 CYP4A11中的翻译后调节。接下来，CYP4A11变体蛋白之间的关系 浓度和蛛网膜酸代谢活性将在人类微粒体和A中进行研究 转基因小鼠模型。这些实验旨在进一步阐明CYP4A11变体在 高血压。 20-HETE是高血压中的重要组成部分。改变其生产的遗传变异 导致高血压敏感性和治疗反应的显着变化。了解 与CYP4A11的表型变化有关的机制将有助于获得对对的更深入的了解 不同个体的高血压并重塑这些特定的人如何治疗血压 减少。