MECHANISMS OF ANESTHETIC CARDIOPROTECTION

 描述（由申请人提供）：我们的PPG的中心主题是使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性的机制的基本见解。我们将使用麻醉剂来测试糖尿病动物和人类心肌细胞对缺血再灌注损伤的保护能力，为糖尿病心脏新的心脏保护策略的未来发展奠定基础。我们的假设是，糖尿病通过对心肌细胞的作用破坏心脏保护作用，这些作用既是环境的，也是细胞的。我们的转化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，计算机模拟和模拟麻醉心脏保护中的线粒体和细胞功能，最后，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解决这一主题的不同方面：项目I（PI：Zeljko J. Bosnjak -麻醉学）将利用来自非糖尿病个体和2型糖尿病患者的iPSC的人类心肌细胞沿着威斯康星州医学院开发的2型糖尿病大鼠模型（T2 DNmtFHH或简称T2 DN）。我们的基本假设是，糖尿病和糖脂毒性损害麻醉心脏保护，通过信号成分，可以有利地调制恢复麻醉心脏保护在糖尿病。项目II（PI：Mingyu Liang - Physiology）将研究miR-21在非糖尿病和糖尿病小鼠和大鼠的麻醉心脏保护中的作用，使用患者特异性心肌细胞将研究结果转化为人类，并研究相关的分子机制。该项目将测试miR- 21表达的变化将恢复糖尿病动物模型和患者特异性心肌细胞中异氟烷赋予的保护作用的假设。项目三（主要研究者：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，并使用测量数据进行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特定作用和相关机制，导致心脏保护，以及糖尿病状况如何损害这种保护。行政核心（主任：Zeljko J. Bosnjak -麻醉学）将为所有子项目和核心项目提供有组织的综合支持框架，以促进项目的内部功能，同时确保合规性。干细胞核心（主任：斯蒂芬A。邓肯-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细胞系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，将由合作多年的研究人员领导，他们的合作历史已经导致了紧密联系的个人项目，非常适合继续这个PPG。这些研究将得到麻醉学研究以及生理学部门的最先进设施的支持;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。

 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.