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（Aldo）激活的受体，调节基因转录 （基因组）和细胞信号传导（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， 心肌梗死（MI）和中风的高风险，MR拮抗剂的保护作用与BP不成比例 降低，支持肾脏独立机制。心肌梗死和中风是由心肌破裂和血栓形成引起的。 发炎的动脉粥样硬化斑块。相对于男性，女性可免受MI和卒中的影响， 绝经期，从而牵连另一种激素，雌激素（E2），而肥胖妇女代谢综合征 在这个不断增长的人口中，随着死亡率的上升，失去了绝经前的保护。我们最近发现， 在雄性小鼠动脉粥样硬化中，内皮细胞（EC）中的白细胞通过诱导白细胞减少而导致血管炎症 募集到血管和表达ICAM 1和E-选择素（E-sel），这在女性中被阻止。我们 还表明雌激素受体α（ER α）与EC中的MR相互作用，阻断ICAM 1的MR调节 并且EC-MR-KO保护肥胖女性免于EC功能障碍。新的数据表明，ICAM 1的表达是 与男性相比，女性的原代人EC减少，这在肥胖女性中逆转; EC-MR 抑制E2的非基因组效应以增加一氧化氮（NO）;并且髓样（My）-MR-KO降低 ApoE-KO小鼠中的斑块大小和炎症，通过活体显微镜（IVM）进行的白细胞滚动，以及 EC粘附配体的巨噬细胞表达。基于这些数据，我们提出检验假设， EC-和My-MR协调动脉粥样硬化中的血管炎症，女性受基因组和非基因组的保护。 MR和ER α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， 包括具有MR、ER α或两者的EC特异性缺失和MR的骨髓特异性缺失的小鼠，每种小鼠都交叉 易患动脉粥样硬化的遗传小鼠模型;肠系膜IVM测定，以测试白细胞运输中的性别差异; 具有ER α功能突变体的EC系;以及年龄匹配的男性和女性人主动脉EC的生物库。使用 这些工具，我们提出了2个具体的目标，每一个使用在体外和体内的方法：SA 1探索基因组 以及MR和ER α在EC中相互作用以调节白细胞募集的非基因组机制， 斑块炎症及其如何介导性别差异; SA 2决定My-MR在免疫细胞中的作用 运输，斑块巨噬细胞表型和骨髓T细胞相互作用的性别差异。使用这些 创新的模型，方法和试剂量身定制，以研究性别差异，这项建议调查的小说 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 和动脉粥样硬化中的血管炎症。目标的完成将确定增加的机制 MR活动性升高的不断增长的人群中的心血管（CV）风险和CV疾病的性别差异。

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.