The Role of MicroRNA-21 in Regulating the Coronary Microcirculation in Diabetes

项目摘要 冠状动脉微血管功能障碍（CMD）与冠状动脉疾病（CAD）、糖尿病性心肌病、 和HFpEF（射血分数保留的心力衰竭）。在健康成人中，一氧化氮（NO）介导 内皮依赖性扩张，而过氧化氢（H2 O2）是患者的主要内皮扩张剂 在CAD这种转变可能是CAD进展中的早期致病步骤，但知之甚少 关于这种转换背后的机制，以及在慢性条件下这种转换是否可以逆转。 该提案通过使用临床前模型来解决NO到H2 O2转换的潜在机制， Gutterman小组的临床CAD观察结果。我们假设“正常”冠状动脉 微血管功能（恢复内皮依赖性舒张）可以通过调节 microRNA-21（miR-21）表达以及miR-21调节内皮（EC）和内皮祖细胞 细胞（EPC）在糖尿病中的作用。为了验证这一假设，我们提出了三个具体的目标，利用遗传 miR-21缺失策略，以解决糖尿病HFHS模型中miR-21缺陷的影响。要求1 通过单细胞RNA测序解决了EPC功能和信号通路的关键方面。目标2利用 心肌内移植EPCs和心肌血流测量，以确定EPCs是否可以 将NO转换为H2 O2。目的3研究miR-21在EC中的特异性作用，以确定miR-21对EC的影响。 预防/逆转NO向H2 O2转换，并评估血管功能和心肌血流量， 糖尿病总的来说，目前的提案将血管生理学概念和方法与状态相结合， 的遗传工具和仪器，以获得新的洞察力的机制，CMD在糖尿病的研究 EPC-EC-微血管功能轴以及围绕miR-1的生理和病理后果 21. 为了确保科学的严谨性，研究质量和职业发展，该奖学金的培训计划 需要将技术技能的获得和同行，同事， 和教授，以促进批判性思维和科学沟通技巧。研究环境在 东北俄亥俄州医科大学可以被描述为高度协作，道德和精通。

PROJECT SUMMARY Coronary microvascular dysfunction (CMD) is linked to coronary artery disease (CAD), diabetic cardiomyopathy, and HFpEF (heart failure with preserved ejection fraction). In healthy adults, nitric oxide (NO) mediates endothelium-dependent dilation, whereas hydrogen peroxide (H2O2) is the principal endothelial dilator in patients with CAD. Such a transition might be an early pathogenic step in the progression of CAD, but little is known about the mechanisms underlying this switch, and whether under chronic conditions the switch can be reversed. This proposal addresses the mechanisms underlying the NO to H2O2 switch by using preclinical models that recapitulate the clinical CAD observations by the Gutterman group. We hypothesize that “normal” coronary microvascular function (restoring endothelial-dependent dilation) can be restored by modulating microRNA-21 (miR-21) expression and that miR-21 regulates endothelial (EC) and endothelial progenitor cell (EPC) function in diabetes. To test this hypothesis, we proposed three specific aims that utilize genetic miR-21 deletion strategies to address the impact of the miR-21 deficiency in a HFHS model of diabetes. Aim 1 addresses key aspects of EPC function and signaling pathways via single cell RNA sequencing. Aim 2 utilizes intramyocardial transplantation EPCs and myocardial blood flow measurements to determine if EPCs can reverse the NO to H2O2 switch. Aim 3 investigates the role of miR-21 specifically in ECs to determine the impact on preventing / reversing the NO to H2O2 switch and assesses vasomotor function and myocardial blood flow in diabetes. Overall, the current proposal merges vascular physiological concepts and methodologies with state- of-the genetic tools and instrumentation to gain novel insight into the mechanisms of CMD in diabetes by studying the EPC-EC-microvascular function axis and the physiological and pathological consequences surrounding miR- 21. To ensure scientific rigor, quality of research, and career development, the training plan laid out for this fellowship entails a combination of technical skill acquisition and regular scientific engagement among peers, colleagues, and professors to facilitate critical thinking and scientific communication skills. The research environment at Northeast Ohio Medical University can be described as highly collaborative, ethical, and proficient.