The Role of Vascular MR-Regulated Genes in Vascular Function and Disease

盐皮质激素受体(Mr)是一种醛固酮(aldosterone，aldo)激活的受体，调节基因转录。 (基因组)和细胞信号(非基因组)来控制细胞功能和调节血压(BP)。先生 在顽固性高血压、肥胖症、心力衰竭和老年人中，激活增加。Aldo预测增加 心肌梗死(MI)和中风的高风险和MR拮抗剂的保护作用与BP不成比例 降低、支持肾脏非依赖性机制。心肌梗塞和中风是由心脏破裂和血栓形成引起的 发炎的动脉粥样硬化斑块。与男性相比，女性受到保护，不会发生心肌梗塞和中风，直到 更年期，从而牵涉到另一种荷尔蒙--雌激素(E2)，而患有代谢综合征的肥胖女性 在这个不断增长的人口中，随着死亡率的上升，人们失去了绝经前的保护。我们最近发现， 内皮细胞(EC)通过诱导白细胞参与雄性小鼠动脉粥样硬化中的血管炎症 血管募集和ICAM1和E-选择素(E-sel)的表达在女性中是被阻止的。我们 也显示雌激素受体α(Era)与内皮细胞中的mR相互作用，阻断mr对ICAM1的调节。 而EC-MR-KO可以保护肥胖女性免受EC功能障碍的影响。新数据显示，ICAM1的表达是 与男性相比，女性的初级人类内皮细胞减少，而肥胖女性的情况正好相反；EC-MR 抑制E_2的非基因组效应增加一氧化氮(NO)；而髓系(MY)-MR-KO减少 载脂蛋白E-KO小鼠的斑块大小和炎症，活体显微镜(IVM)白细胞滚动，以及 巨噬细胞表达EC黏附配体。基于这些数据，我们建议检验这一假设 EC-和MY-MR协调动脉粥样硬化中的血管炎症，女性受到基因组和非基因组的保护 Mr和era之间的基因组相互作用。为了测试这一点，我们创造了创新的试剂和方法 包括EC特异性MR、ERA或两者缺失以及髓系特异性MR缺失的小鼠，每个交叉 易患动脉粥样硬化的遗传性小鼠模型；肠系膜IVM试验以测试白细胞转运的性别差异； 具有ERA功能突变的EC系；以及年龄匹配的男性和女性人主动脉内皮细胞的生物库。vbl.使用 这些工具我们提出了两个特定的目标，每个都使用了体外和体内的方法：SA1探索基因组 以及MR和ERA在内皮细胞中相互作用以调节白细胞募集和 斑块炎症及其如何调节性别差异；SA2决定My-MR在免疫细胞中的作用 髓系T细胞相互作用中的运输、斑块巨噬细胞表型和性别差异。使用这些 为研究性别差异而量身定做的创新模型、方法和试剂，这项提案调查了这部小说 EC-和MY-MR协调血管和免疫细胞反应以诱导白细胞募集的概念 动脉粥样硬化中的血管炎症。目标的完成将确定增加的潜在机制 MR活性升高和心血管疾病的性别差异在不断增长的人群中的心血管风险。

The mineralocorticoid receptor (MR) is an aldosterone (Aldo)-activated receptor that regulates gene transcription (genomic) and cellular signaling (non-genomic) to control cell functions and regulate blood pressure (BP). MR activation is increased in resistant hypertension, obesity, heart failure and the elderly. Increased Aldo predicts high risk of myocardial infarction (MI) and stroke and MR antagonists are protective out of proportion to BP lowering, supporting kidney-independent mechanisms. MI and stroke are caused by rupture and thrombosis of inflamed atherosclerotic (athero) plaques. Women are protected from MI and stroke relative to men until menopause thereby implicating another hormone, estrogen (E2), while obese women with metabolic syndrome lose premenopausal protection with rising mortality in this growing demographic. We recently showed that MR in endothelial cells (EC) contributes to vascular inflammation in athero in male mice by inducing leukocyte recruitment to vessels and expression of ICAM1 and E-selectin (E-sel) and that this is prevented in females. We also showed that estrogen receptor alpha (ERa) interacts with the MR in ECs, blocking MR regulation of ICAM1 and that EC-MR-KO protects obese females from EC dysfunction. New data shows that ICAM1 expression is decreased in primary human ECs from females vs males and this is reversed in obese females; that EC-MR inhibits the non-genomic effect of E2 to increase nitric oxide (NO); and that myeloid (My)-MR-KO decreases plaque size and inflammation in ApoE-KO mice, leukocyte rolling by intravital microscopy (IVM), and macrophage expression of EC adhesion ligands. Based on these data, we propose to test the hypothesis that EC- and My-MR coordinate vascular inflammation in athero and that females are protected by genomic and non- genomic interactions between MR and ERa. To test this, we have created innovative reagents and methods including mice with EC-specific deletion of MR, ERa or both and myeloid-specific deletion of MR, each crossed to athero-prone genetic mouse models; a mesenteric IVM assay to test sex differences in leukocyte traficking; EC lines with ERa functional mutants; and a biobank of age matched male and female human aortic ECs. Using these tools we propose 2 specific aims, each using in vitro and in vivo approaches: SA1 explores the genomic and non-genomic mechanisms by which MR and ERa interact in ECs to regulate leukocyte recruitment and plaque inflammation and how this mediates sex differences; SA2 determines the role of My-MR in immune cell trafficking, plaque macrophage phenotype, and sex-differences in myeloid-T cell interactions. Using these innovative models, methods, and reagents tailored to study sex-differences, this proposal investigates the novel concept that EC- and My-MR coordinate vascular and immune cell responses to induce leukocyte recruitment and vascular inflammation in athero. Completion of the aims will identify mechanisms underlying increased cardiovascular (CV) risk in growing populations with elevated MR activity and for sex differences in CV disease.