The Role of 12-Lipoxygenase in Aldosterone Secretion

12-脂氧合酶在醛固酮分泌中的作用

基本信息

批准号：
8957498
负责人：
Phillip G Kopf
金额：
$ 41.78万
依托单位：
MIDWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8957498
关键词：
12-HETE Acids Adrenal Glands Affinity Aldosterone Alzheimer&apos s Disease Angiotensin II Arachidonate 12-Lipoxygenase Arachidonic Acids Atherosclerosis Binding Calcium Calcium Signaling Cardiovascular Diseases Cell membrane Cells Congestive Heart Failure Corticotropin Cyclic AMP Data Diabetes Mellitus Disease Fostering G-Protein-Coupled Receptors GTP-Binding Proteins Goals Health Heart Hypertrophy Hydroxyeicosatetraenoic Acids Hyperaldosteronism Hyperplasia Hypertension Inflammatory Laboratories Malignant Neoplasms Measures Mediating Membrane Methods Mission Morbidity - disease rate Outcome Parkinson Disease Pathway interactions Patients Phospholipase C Potassium Production Public Health Receptor Activation Receptor, Angiotensin, Type 1 Regulation Research Role Signal Transduction Stimulus Testing Thrombosis Work Zona Glomerulosa adrenic acid analog base functional group guanine nucleotide binding protein improved inhibitor/antagonist innovation liquid chromatography mass spectrometry mortality novel novel therapeutic intervention public health relevance receptor receptor coupling remediation

项目摘要

DESCRIPTION (provided by applicant): Evidence exists for an essential role of the 12-lipoxygenase (12-LO) pathway in Ang II-stimulated aldosterone secretion. However, the identity of 12-LO metabolites, as well as the mechanism and extent by which these metabolites contribute to Ang II-stimulation of aldosterone secretion remains unknown. The long-term goal of these studies is to understand the role of 12-LO in the regulation of aldosterone secretion. The objective of this proposal is to identify the 12-LO metabolites that contribute to Ang II-stimulated aldosterone secretion and to characterize the receptor that mediates the action of these active metabolites. The central hypothesis is that arachidonic acid (AA)-derived hydroxyeicosatetraenoic acids (HETEs) and/or adrenic acid (AdA)-derived hydroxydocosatetraenoic acids (HDTEs) mediate Ang II-stimulated aldosterone secretion by activation of a guanine nucleotide binding protein (G-protein) coupled receptor (GPCR). This hypothesis has been formulated on the basis of preliminary data produced in the applicant's laboratory. The rationale for the proposed research is that identification of the 12-LO metabolites and their receptor that mediates aldosterone secretion would provide a novel pharmacological target for the remediation of circulating aldosterone levels. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Identify the 12-LO metabolites that contribute to aldosterone secretion; and 2) Characterize the receptor that mediates the activity of 12-LO metabolites in ZG cells. Under the first aim, 12-LO AA and AdA metabolites that increase following these steroidogenic stimuli will be identified by liquid chromatography/mass spectrometry (LC/MS), and then these metabolites will be tested for steroidogenic activity. Under the second aim, the ability and potency of these 12-LO AA and AdA metabolites to activate a GPCR on ZG membranes will be tested by GTPγ[35S] binding, as well as their ability to increase cAMP and intracellular calcium. Additionally, the type of Ga subunit that is activated by these metabolites will also be determined. Furthermore, analogs of 12-HETE and 14- HDTE will be used to determine the functional groups necessary for both receptor activation and calcium signaling. The approach is innovative, because it will develop specific LC/MS methods to measure adrenal HETEs and HDTEs and define the role of both AA and AdA metabolites in aldosterone production. The proposed research is significant, because the characterization of a 12(S)-HETE and/or 14(S)-HDTE receptor is an essential step towards understanding the critical role of 12-LO metabolites in the regulation of aldosterone secretion. Ultimately, characterization of this novel GPCR will provide an innovative pharmacological target to not only ameliorate cardiovascular disease associated with hyperaldosteronism, but potentially many other diseases in which 12-LO metabolites have been implicated including inflammatory, hyperproliferative, and hyperplastic diseases such as atherosclerosis, diabetes, Parkinson's disease, Alzheimer's disease, and cancer.

描述（由适用提供）：存在证据证明ANG II刺激的醛固酮分泌中12-脂氧合酶（12-LO）途径的重要作用。然而，这些代谢产物有助于醛固酮分泌的ANG II刺激的机制和程度仍然未知。这些研究的长期目标是了解12-LO在醛固酮分泌调节中的作用。该提案的目的是确定有助于ANG II刺激的醛固酮分泌的12-LO代谢产物，并表征介导这些活性代谢产物作用的接收器。中央假设是蛛网膜酸（AA）衍生的羟基羟基乙烯酸（HETES）和/或肾上酸（ADA）衍生的羟基核酸乙烯酸（HDTES）介导Ang Ang II-II刺激的AldostoRERORORTION ANDDOSTORETION of ATEPTINANE RIDEROTION（GUIDERER）的蛋白酶（Gried ot）cprote蛋白酶（hdtes）（GPCR）。该假设是根据应用程序实验室产生的初步数据提出的。拟议研究的理由是，介导醛固酮分泌的12-LO代谢产物及其受体的鉴定将为修复循环醛固酮水平的补救提供新的药物靶标。在强大的初步数据的指导下，将通过追求两个具体目标来检验该假设：1）确定有助于醛固酮分泌的12-LO代谢物； 2）表征介导ZG细胞中12-LO代谢产物活性的接收器。在第一个目标下，将通过液相色谱/质谱法（LC/MS）鉴定出12-LO AA和ADA代谢产物，这些代谢物将在这些类固醇生成刺激后增加，然后将测试这些代谢物的类固醇生成活性。在第二个目标下，这些12-LO AA和ADA代谢产物在ZG膜上激活GPCR的能力和效力将由GTPγ[35S]结合进行测试，以及它们增加cAMP和细胞内钙的能力。另外，还将确定这些代谢产物激活的GA亚基的类型。此外，将使用12-HETE和14-HDTE的类似物来确定受体激活和钙信号传导所必需的官能团。该方法具有创新性，因为它将开发特定的LC/MS方法来测量肾上腺het和HDTE，并定义AA和ADA代谢产物在醛固酮生产中的作用。拟议的研究很重要，因为12（s） - hete和/或14（s）-HDTE受体的表征是理解12-LO代谢产物在醛固酮分泌调节中的关键作用的重要步骤。最终，这种新型GPCR的特征将提供创新的药物靶标，不仅可以改善与高醛烷主义相关的心血管疾病，而且可能暗示了12-lo代谢产物的许多其他疾病，包括炎症性，增生性增殖性超增长和增生性疾病，例如动脉粥样硬化，糖尿病性疾病，parkinson和parkinson and and and and and and and and and and and and and and and and and and and and and andzenshonshesson，综合疾病。