Pyruvate Dehydrogenase Complex Activation as a Strategy to Ameliorate Metabolic Disease

代谢重塑是心脏病、2型糖尿病等疾病的一个潜在主题， 糖尿病和癌症，其中细胞偏离其典型的燃料利用概况。丙酮酸 脱氢酶复合体（PDC）是有氧葡萄糖利用的看门人， 下调与这些疾病密切相关。因此，最近的努力集中在 作为一种治疗剂的PDC的激活。目前，努力的重点仅仅是抑制 丙酮酸脱氢酶激酶（PDK），其通过磷酸化抑制PDC。然而，在这方面， 激活丙酮酸脱氢酶磷酸酶（PDP）作为一种治疗剂， 磷酸化PDC以恢复活性，已被忽略。此外，众所周知， PDC活性受到NADH的抑制，但针对这种调节机制进行治疗的努力 被忽视了作为一个更全面的PDC激活策略，我们希望扩大 PDK抑制剂，鉴定PDP小分子活化剂，并优化醌化合物， 已经显示出从NADH抑制中恢复PDC作为一种新的治疗策略， 代谢性疾病第二，线粒体产生的活性氧（ROS） 电子传递链（mETC）被认为是增强PDC活化，但影响 PDC作为ROS的来源以及特定条件如NADH/NAD和ATP需求， 在运动中，影响ROS稳态尚不清楚。ROS的位点特异性定量 生成需要在各种条件下进行详细的酶模拟来阐明。的 该项目的目的是1）虚拟筛选和实验验证PDC活化剂，2） 应用数学建模来确定PDC活化对位点特异性mETC ROS的影响 一代我们从ZINC数据库中获得了1500万个药物样虚拟化合物， 在三个已知抑制位点：硫辛酰胺，ATP/ADP， 和PZF 3来寻找同工酶特异性和PAN抑制剂。这个虚拟复合集也将被用于 通过虚拟筛选混合晶体/计算衍生的PDPc来识别PDP活化剂， PDPr复合物。我们将使用PDC（E3）的NADH结合结构域，以结构引导的方式， 方法，以确定PDC的最佳醌类似物。详细的酶动力学模型 PDC、TCA循环和mETC将被整合以模拟各种氧化状态，包括 还原性应激和运动条件，以量化位点特异性线粒体ROS 生产我们认为，我们的建议解决了在激活 PDC作为一种治疗，并将提供一个定量的了解PDC的影响， 线粒体ROS的产生。

Metabolic remodeling is an underlying theme in diseases such as heart disease, type 2 diabetes, and cancer, where cells deviate from their typical fuel utilization profile. The pyruvate dehydrogenase complex (PDC) is the gatekeeper for aerobic glucose utilization and its downregulation is greatly associated with these diseases. As such, recent efforts have focused on the activation of PDC as a therapeutic. Currently, efforts have focused only on inhibition of the pyruvate dehydrogenase kinase (PDK), which inhibits PDC by phosphorylation. However, activation of the pyruvate dehydrogenase phosphatase (PDP) as a therapeutic, which de- phosphorylates PDC to recover activity, has been ignored. Furthermore, it is well-known that PDC activity is inhibited by NADH but efforts to therapeutically target this regulatory mechanism have been overlooked. As a more comprehensive PDC activation strategy, we look to expand PDK inhibitors, identify PDP small molecule activators, and optimize quinone compounds that have been shown to recover PDC from NADH inhibition as a new therapeutic strategy for metabolic disease. Secondly, reactive oxygen species (ROS) generated from the mitochondrial electron transport chain (mETC) are thought to be enhanced by PDC activation but the impact of PDC as a source of ROS and how specific conditions such as NADH/NAD and ATP demand, as in exercise, influence ROS homeostasis is unclear. Site-specific quantification of ROS generation requires detailed enzymatic simulations in various conditions to be elucidated. The aims of this project are to 1) virtually screen and experimentally validate PDC activators and 2) apply mathematical modeling to determine effects of PDC activation on site-specific mETC ROS generation. We obtained 15M drug-like virtual compounds from the ZINC database to individually screen all PDK isozymes (1-4) at three known inhibition sites: lipoamide, ATP/ADP, and pzf3 to find isozyme specific and pan inhibitors. This virtual compound set will also be used to identify PDP activators by virtually screening a hybrid crystal/computationally derived PDPc- PDPr complex. We will use the NADH binding domain of PDC (E3), in a structure guided approach, to identify optimal quinone analogs for PDC. Detailed enzyme kinetics models of PDC, TCA cycle, and mETC will be integrated to simulate various oxidative states including reductive stress and exercising conditions to quantify site-specific mitochondrial ROS production. We believe that our proposal addresses significant gaps in strategies to activate PDC as a therapeutic and will provide a quantitative understanding of the influence of PDC activity on mitochondrial ROS production.