MECHANISMS OF ANESTHETIC CARDIOPROTECTION

 描述(申请人提供)：我们PPG的中心主题是利用患者来源的心肌细胞和糖尿病动物获得对糖尿病心脏对缺血-再灌注损伤更敏感的机制的基本见解。我们将使用麻醉药来测试糖尿病动物和人类心肌细胞抵抗缺血再灌注损伤的能力，为未来开发新的糖尿病心脏保护策略奠定基础。我们的假设是，糖尿病通过对起源于环境和细胞的心肌细胞的作用来破坏心脏保护。我们的翻译方法将包括体内糖尿病动物模型、分离的动物心脏制剂、分离的动物心肌细胞和线粒体、麻醉心脏保护中线粒体和细胞功能的计算机模拟和建模，最后是来自疾病特异性诱导多能干细胞(IPSC)的人类心肌细胞。以下密切相关和相互依存的项目将涉及这一主题的不同方面：项目I(PI：Zeljko J.Bosnjak-麻醉学)将利用从非糖尿病患者和2型糖尿病患者的IPSC获得的人心肌细胞以及威斯康星医学院建立的2型糖尿病大鼠模型(T2DNmtFHH或T2DN)。我们的基本假设是，糖尿病和糖脂毒性通过信号成分损害麻醉性心脏保护，这些信号成分可以在糖尿病期间被有利地调节以恢复麻醉性心脏保护。项目II(PI：梁明宇-生理学)将研究miR-21在非糖尿病和糖尿病小鼠和大鼠麻醉心脏保护中的作用，使用患者特定的心肌细胞将研究结果转化为人类，并探讨相关的分子机制。该项目将检验一种假设，即改变miR-21的表达将恢复异氟醚对糖尿病动物模型和患者特定心肌细胞的保护作用。项目III(PI：Ranjan K.Dash-生物技术和生物工程中心和生理学)将使用系统生物学方法反复进行实验，并使用测量数据来计算建模和机械表征挥发性麻醉剂对线粒体和细胞功能的具体影响和相关机制，从而导致心脏保护，以及糖尿病条件如何损害这种保护。行政核心(主任：Zeljko J.Bosnjak-麻醉学)将为所有子项目和核心提供有组织和全面的支持框架，有助于该计划的凝聚力，同时确保法规遵从性。干细胞核心公司(董事：Stephen A.Duncan-细胞生物学、神经生物学和解剖学)将通过提供患者来源的IPSC株并将这些细胞分化为疾病特异性心肌细胞来支持所有项目。因此，所有三个项目都集中在糖尿病破坏心脏保护的作用上。这是一个高度集中的项目，将由合作多年的调查人员领导，他们的合作历史导致了紧密联系的个别项目，这些项目非常适合继续开展这一项目。这些研究将得到麻醉学研究、生理学、生物物理学、生物化学、药理学、细胞生物学、神经生物学和解剖学、医学、心血管研究中心、人类和分子遗传中心以及生物技术和生物工程中心等先进设施的支持。

 DESCRIPTION (provided by applicant): The central theme of our PPG is to gain fundamental insight into the mechanisms responsible for greater susceptibility of diabetic hearts to ischemia-reperfusion injury using both patient-derived cardiomyocytes and diabetic animals. We will use anesthetics to test the ability of diabetic animals and human cardiomyocytes to be protected against ischemia-reperfusion injury for the future development of novel cardioprotective strategies for the diabetic heart. Our hypothesis is that diabetes undermines cardioprotection through actions on cardiomyocytes that are both environmental and cellular in origin. Our translational approach will include in vivo diabetic animal models, isolated animal heart preparations, isolated animal cardiomyocytes and mitochondria, computer simulations and modeling of mitochondrial and cellular function in anesthetic cardioprotection, and finally, human cardiomyocytes derived from disease-specific induced pluripotent stem cells (iPSC). The following closely interrelated and interdependent Projects will address different facets of this theme: Project I (PI: Zeljko J. Bosnjak - Anesthesiology) will utilize human cardiomyocytes derived from the iPSC obtained from non-diabetic individuals and patients with type 2 diabetes mellitus along with a rat model of type 2 diabetes developed at the Medical College of Wisconsin (T2DNmtFHH or T2DN for short). Our fundamental hypothesis is that diabetes and glucolipotoxicity impair anesthetic cardioprotection through signaling components that can be favorably modulated to restore anesthetic cardioprotection during diabetes. Project II (PI: Mingyu Liang - Physiology) will examine the role of miR-21 in anesthetic cardioprotection in non-diabetic and diabetic mice and rats, translate the findings to human using patient-specific cardiomyocytes, and investigate the molecular mechanisms involved. This project will test the hypothesis that a change of miR- 21 expression will restore isoflurane-conferred protection in diabetic animal models and in patient-specific cardiomyocytes. Project III (PI: Ranjan K. Dash - Biotechnology and Bioengineering Center and Physiology) will use a system biology approach to iteratively conduct experiments and use the measured data to computationally model and mechanistically characterize the specific effects and associated mechanisms of volatile anesthetic action on mitochondrial and cellular function that lead to cardioprotection, and how diabetic conditions impair this protection. Administrative Core (Director: Zeljko J. Bosnjak - Anesthesiology) will provide an organized and comprehensive framework of support for all subprojects and cores contributing to the cohesive functionality of the Program while ensuring regulatory compliance. Stem Cell Core (Director: Stephen A. Duncan - Cell Biology, Neurobiology and Anatomy) will support all projects by supplying patient-derived iPSC lines and differentiate these cells into disease-specific cardiomyocytes. Thus, all three Projects focus cohesively on the role of diabetes undermining the cardioprotection. This is a highly focused Program that will be led by investigators who have worked together for many years, and their history of collaboration has resulted in closely linked individual Projects that are ideally suitedfor the continuation of this PPG. These investigations will be supported by well-established state-of-the-art facilities in Anesthesiology Research, and the Departments of Physiology; Biophysics; Biochemistry; Pharmacology; Cell Biology, Neurobiology and Anatomy; Medicine; Cardiovascular Research Center; Human and Molecular Genetic Center and Biotechnology and Bioengineering Center.