Cav-3 in Diabetic Myocardial Injury Following Ischemia/Reperfusion

肥胖/糖尿病对心肌缺血/再灌注（MI/R）损伤的不利影响 机制等内脏脂肪细胞（ADp）功能障碍导致远端器官损伤。然而，分子 功能障碍性ADp和MI/R损伤增加之间的联系尚未确定。细胞外囊泡， 特别是外来体（exosomes，Exo），是介导器官间通讯的系统性信使。但无论 以及糖尿病如何改变Exo介导的ADP-心脏通讯仍然未知。我们的初步 实验表明，糖尿病会导致供体（ADp）和供体（ADp）发生显着的病理变化， 受体（心肌细胞，CM）细胞，增加MI/R损伤。这个应用程序将测试一个假设，糖尿病 Exo介导的ADp与心脏通讯的改变是加重MI后心脏损害的新机制。 重塑将讨论三个具体目标。SA 1将澄清下游机制， 糖尿病ADp Exo急性MI/R损伤加重。我们首次发现miR-130 b-3 p是一种常见的 在糖尿病患者血清和糖尿病患者血清中，ADp Exo分子显著增加。MiR-130 b-3 p模拟物 miR 130 b-3 p抑制剂加重了MI/R损伤，而miR 130 b-3 p抑制剂减轻了急性MI/R损伤。这方面的实验将 1)将ADp定义为糖尿病心脏中miR-130 b-3 p的细胞来源;以及2）鉴定ADp的下游表达。 介导miR-130 b-3 p促凋亡作用的分子靶点。SA 2将确定负责的机制 增加糖尿病CM对ADp Exo的摄取。初步数据表明，Cav 3表达减少， 糖尿病心脏中Cav 3硝化的增加可能是ADp Exo跨膜丢失的原因 通过CM的ADp Exo摄取的信号传导和促进。结合遗传学和药理学方法，我们将 严格测试一个新的假设，Cav 3/AdipoR 1信号复合物的完整性是决定命运的关键， ADp Exo的。糖尿病心脏Cav 3/AdipoR 1的解离将AdipoR 1从介导ADp的受体转换， 衍生的富含脂联素的Exo在非糖尿病CM中启动信号传导至促进糖尿病ADp Exo的媒介物 进入CM。SA 3将检验阻断Exo介导的ADp-CM通讯的干预措施的假设 是治疗糖尿病心肌梗死后重构和心力衰竭的新疗法。初步数据显示， 心肌内注射糖尿病性ADpExo可加重非糖尿病小鼠的急性MI/R损伤，而 Exo产生抑制减轻糖尿病小鼠的MI/R损伤这一目标的实验将确定是否 ADP特异性miR-130 b-3 pKO或CM特异性Cav 3 OE可有效保护糖尿病心脏免于过度表达。 心肌梗死后心脏重塑为了增加我们研究结果的转化价值，我们将确定是否 miR-130 b-3 p抑制剂或过氧亚硝酸盐分解催化剂的施用有效地保护了抗 糖尿病ADpExo介导的心脏重塑和HF增强。成功完成这些研究将 揭示糖尿病ADp Exo诱导的心脏损伤的分子机制，产生干预措施 靶向Exo介导的ADp-CM通讯，并最终保护心脏免受糖尿病MI/R损伤。

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.