Mineralocorticoid receptor mediates vascular stiffness via dysregulated immunity in perivascular adipose tissue

项目摘要 我的研究主要集中在血管周围的盐皮质激素受体（MR）信号之间的相互作用， 脂肪组织（PVAT），免疫失调和血管功能障碍，在高脂高蛋白背景下， 果糖饮食（西方饮食[WD]）。WD导致胰岛素抵抗。重要的是，胰岛素抵抗导致 女性的侵袭性心血管疾病。我假设在女性中，WD诱导的胰岛素抵抗 导致T调节（Treg）功能丧失，导致巨噬细胞MR活化和随后的M1 PVAT中的极化。此外，我假设这些事件通过激活 组织转氨酶-2（TG 2），部分通过氧化应激和生物可利用的一氧化氮减少 （否）。为了验证我的假设，具体目标1是确定PVAT巨噬细胞中MR激活的效果 关于血管僵硬的发病机制，因为它与Treg功能受损和M1巨噬细胞有关 极化骨髓特异性MR敲除（MyMRKO）雄性和雌性小鼠将用WD或 我们会测量主动脉硬度在PVAT中，我们将测量巨噬细胞极化， 炎症我们将测量原代内皮细胞（EC）和血管平滑肌的细胞硬度 在PVAT条件培养基中培养的VSMC。在具体目标2中，我们将确定 TdR对PVAT巨噬细胞极化和主动脉僵硬度的作用。在这组实验中，我们将使用 从喂食正常饮食的雄性和雌性C57 B1/6 J小鼠向用WD或D2 D处理的小鼠的过继性Treg转移 缓慢升压剂量的醛固酮。我们将测量巨噬细胞极化，炎症和主动脉 刚度，沿着在PVAT调节介质中进行EC/VSMC刚度测量。我已经召集了一个团队 导师和合作者，他们将在整个项目中指导我，并促进我作为一个 独立研究员。我的职业发展计划有一个初始培训期（1-3年）， 完成建议的工作，以及一个过渡期（4-5年），在此期间，我将专注于R 01建议。 我的指导团队拥有必要的基础设施、专业知识和资源，我的机构将提供 必要的支持，使我能够成功地实现我的目标。我的中期目标是 完成我提出的工作。我的最终目标是建立一个明确的工作路线， MR激活和免疫失调导致血管疾病。

Project summary My research is focused on the interactions between mineralocorticoid receptor (MR) signaling in perivascular adipose tissue (PVAT), dysregulation of immunity and vascular dysfunction in the setting of high-fat high- fructose diet (Western Diet [WD]). WD leads to insulin resistance. Importantly, insulin resistance leads to aggressive cardiovascular disease in females. I hypothesize that in females, WD-induced insulin resistance results in loss of T regulatory (Treg) function leading to macrophage MR activation and subsequent M1 polarization in PVAT. Further, I hypothesize that these events increase vascular stiffness via activation of Tissue transglutaminase-2 (TG2), in part, through oxidative stress and decreased bioavailable nitric oxide (NO). To test my hypothesis, specific Aim 1 is to determine the effect of MR activation in PVAT macrophages on the pathogenesis of vascular stiffness as it relates to impaired Treg function and M1 macrophage polarization. Myeloid-specific MR knockout (MyMRKO) male and female mice will be treated with a WD or aldosterone and we will measure aortic stiffness. In PVAT, we will measure macrophage polarization and inflammation. We will measure cellular stiffness in primary endothelial cells (EC) and vascular smooth muscle cells (VSMC) cultured in PVAT-conditioned media from each cohort. In Specific Aim 2, we will determine the role of Tregs on PVAT macrophage polarization and aortic stiffness. In this set of experiments, we will use adoptive Treg transfer from male and female C57Bl/6J mice fed a normal diet to the mice treated with WD or slow pressor doses of aldosterone. We will measure macrophage polarization, inflammation and aortic stiffness, along with EC/VSMC stiffness measurement in PVAT-conditioned media. I have assembled a team of mentors and collaborators that will guide me throughout this project and promote my transition as an independent researcher. My career development plan has an initial training period (years 1-3) in which I will complete the proposed work, and a transitional period (years 4-5) during which I will focus on an R01 proposal. My mentoring team has the necessary infrastructure, expertise and resources and my institution will provide the necessary support to enable me successfully achieve my goals. My intermediate goal is to successfully complete the work I propose. My ultimate goal is to establish a distinct line of work addressing the mechanisms by which MR activation and dysregulated immunity lead to vascular disease.