Cav-3 in Diabetic Myocardial Injury Following Ischemia/Reperfusion

肥胖/糖尿病对心肌缺血/再灌注损伤的不利影响 机械装置。内脏脂肪细胞(ADP)功能障碍导致远隔器官损伤。然而，分子 ADP功能障碍与MI/R损伤增加之间的联系(S)仍未确定(S)。细胞外小泡， 尤其是Exosome(Exo)，是调节器官间通讯的系统信使。然而，无论是 糖尿病如何改变外周介导的ADP-心脏通讯仍不清楚。我们的预赛 实验表明，糖尿病引起供者(ADP)和供者(ADP)的显著病理改变 受体(心肌细胞，CM)细胞，增加MI/R损伤。这个应用程序将检验一个假设，即糖尿病 外源性ADP向心脏通讯的改变是心肌梗死后心脏恶化的一种新机制 改建。将解决三个具体目标。SA1将澄清负责以下工作的下游机制 糖尿病ADP加重急性MI/R损伤。我们首次揭示了miR-130b-3p是一种常见的 糖尿病ADP Exo和糖尿病患者血清中的分子显著增加。MIR-130B-3P模拟 而miR130b-3p抑制剂可减轻急性MI/R损伤。以此为目标的实验将 1)将ADP定义为糖尿病心脏miR-130b-3p的细胞来源；2)确定下游 MiR-130b-3p促凋亡作用的分子靶点SA2将确定负责的机制 用于糖尿病CM对ADP Exo的摄取增加。初步数据表明，Cav3表达减少和 糖尿病心脏中Cav3硝化增加可能是ADP外膜丢失的原因 CM对ADP外源摄取的信号转导和促进作用结合遗传学和药理学方法，我们将 严格检验一个新的假设，即Cav3/AdipoR1信号复合体的完整性对决定命运至关重要 ADP Exo.糖尿病时心脏Cav3/AdipoR1的解离使AdipoR1从介导ADP-R的受体上切换 衍生的富含脂联素的Exo在非糖尿病CM中向促进糖尿病ADP Exo的载体发出信号 CM中的条目。SA3将检验一种假设，即阻断Exo的干预措施介导了ADP-CM通信 是治疗糖尿病心肌梗死后重塑加重和心力衰竭的新疗法。初步数据显示 心肌内注射糖尿病ADP Exo可加重非糖尿病小鼠的急性MI/R损伤，而 抑制外源性合成减轻糖尿病小鼠的心肌缺血/再灌注损伤。这一目标的实验将决定是否 ADP特异性miR-130b-3pKO或CM特异性Cav3OE在糖尿病心脏保护中的作用 心肌梗死后心脏重塑。为了增加我们发现的翻译价值，我们将确定 使用miR-130b-3p缓蚀剂或过氧亚硝酸根分解催化剂可有效地防止 糖尿病ADP外源性介导的心脏重塑和心衰。成功完成这些研究将 揭示糖尿病ADP外源性心脏损伤的分子机制，产卵干预 靶向Exo介导的ADP-CM通讯，最终对糖尿病心肌缺血再灌注损伤具有保护作用。

Obesity/diabetes adversely impact myocardial ischemia/reperfusion (MI/R) injury by incompletely understood mechanisms. Visceral adipocyte (ADp) dysfunction contributes to remote organ injury. However, the molecular link(s) between dysfunctional ADp and increased MI/R injury remain(s) unidentified. Extracellular vesicles, particularly exosomes (Exo), are systemic messengers mediating inter-organ communication. However, whether and how diabetes may alter Exo-mediated ADp-heart communication remain unknown. Our preliminary experiments demonstrate that diabetes causes significant pathologic alterations in both donor (ADp) and recipient (cardiomyocyte, CM) cells, increasing MI/R injury. This application will test a hypothesis that diabetic alteration of Exo-mediated ADp to heart communication is a novel mechanism exacerbating post-MI cardiac remodeling. Three specific aims will be addressed. SA1 will clarify the downstream mechanisms responsible for diabetic ADp Exo exacerbation of acute MI/R injury. We revealed for the first time that miR-130b-3p is a common molecule significantly increased in diabetic ADp Exo and diabetic patient serum. MiR-130b-3p mimics exacerbated MI/R injury, whereas miR130b-3p inhibitor mitigated acute MI/R injury. Experiments in this aim will 1) define ADp as the cellular source of miR-130b-3p in the diabetic heart; and 2) identify the downstream molecular targets mediating the pro-apoptotic effect of miR-130b-3p. SA2 will identify mechanisms responsible for increased uptake of ADp Exo by diabetic CM. Preliminary data suggest that reduced Cav3 expression and increased Cav3 nitration in the diabetic heart is likely responsible for the loss of ADp Exo transmembrane signaling and promotion of ADp Exo uptake by CM. Combining genetic and pharmacologic approaches, we will rigorously test a novel hypothesis that Cav3/AdipoR1 signaling complex integrity is critical in determining the fate of ADp Exo. Diabetic dissociation of cardiac Cav3/AdipoR1 switches AdipoR1 from a receptor mediating ADp- derived, adiponectin-rich Exo initiated signaling in non-diabetic CM to a vehicle facilitating diabetic ADp Exo entry within CM. SA3 will test a hypothesis that interventions blocking Exo mediated ADp-CM communication are novel therapy against diabetic exacerbation of post-MI remodeling and HF. Preliminary data demonstrate that intramyocardial injection of diabetic ADp Exo exacerbates acute MI/R injury in non-diabetic mice, whereas Exo production inhibition attenuates MI/R injury in diabetic mice. Experiments in this aim will determine whether ADp-specific miR-130b-3pKO or CM-specific Cav3OE is effective in protecting the diabetic heart from excessive post-MI cardiac remodeling. To increase the translational value of our findings, we will determine whether administration of miR-130b-3p inhibitor or peroxynitrite decomposition catalyst effectively protects against diabetic ADp Exo-mediated augmented cardiac remodeling and HF. Successful completion of these studies will reveal the molecular mechanisms responsible for diabetic ADp Exo-induced cardiac injury, spawn interventions targeting Exo mediated ADp-CM communication, and ultimately protect against cardiac diabetic MI/R injury.