Basic and Translational Studies in Redox Regulation of Cardiovascular Physiology and Disease

摘要 我们的研究计划旨在阐明化学，生物学和翻译的影响，“氧化还原开关”， 心血管系统。我们的研究表明，细胞色素b5还原酶3（CYB5R3）作为一种酶， 关键的氧化还原开关在心血管系统通过还原底物，如血红素和辅酶Q， 调节活性氮和氧物质以及下游信号传导。R35的首要目标是 一项研究计划是填补我们在心血管系统中与氧化还原开关相关的知识中的四个主要空白。 生物学首先，我们将定义“其他CYB5R”家族成员的作用，包括CYB5R1、2、4、5及其 在内皮细胞、平滑肌细胞和心肌细胞中起作用。虽然在这些细胞类型中表达，但没有 报告定义了这些酶在心血管系统中的功能。使用新的细胞类型特异性 CYB5R1、2、4、5的敲除，我们将描述这些CYB5R1、2、4、5的生理和病理生理作用。 跨内皮、平滑肌和心肌细胞的酶及其在氧化还原中的潜在功能 发信号。其次，我们的目标是建立新的客户蛋白的CYB5R家族的酶使用创新的 “生物淘选”方法。利用转基因大豆过氧化物酶耦合CYB5R酶，我们将 为这些还原酶建立新的结合伴侣、底物和机制。第三，利用大型人力 数据集（即n=8500名受试者），我们将评估CYB5R酶中的常见遗传变异是否与 患有高血压、镰状细胞病和心力衰竭等疾病。机械研究将决定 这些变体对心血管细胞中氧化还原信号传导途径的功能影响。第四，我们将利用 从我们对CYB5R3的研究中获得的信息，以测试我们开发的一种名为“NitroQ”的新药， if改善氧化还原平衡并逆转高血压、镰状细胞病和心力衰竭。机械论 研究将阐明NitroQ影响的关键靶点和下游信号通路。总的来说，这些 基本的基础，转化和治疗研究将提供一个关键的框架，使我们能够填补 这些主要的差距，我们的知识和提高我们的理解氧化还原开关在心血管健康 和疾病

ABSTRACT Our research program seeks to elucidate the chemistry, biology and translational impact of “redox switches” in the cardiovascular system. Our work has revealed that cytochrome b5 reductase 3 (CYB5R3) operates as a critical redox switch in the cardiovascular system by reducing substrates such heme and coenzyme Q to modulate reactive nitrogen and oxygen species and downstream signaling. The overarching goal of this R35 research proposal is to fill four major gaps in our knowledge related to redox switches in the cardiovascular biology. First, we will define the role(s) of the “other CYB5R” family members including CYB5R1, 2, 4, 5 and their functions in endothelial, smooth muscle and cardiomyocytes. While expressed in these cells types, there are no reports defining the function(s) of these enzymes in the cardiovascular system. Using new cell type specific knockouts of CYB5R1, 2, 4, 5, we will delineate the physiological and pathophysiological role(s) of these enzymes across endothelial, smooth muscle and cardiomyocytes and their potential function(s) in redox signaling. Second, we aim to establish novel client proteins for the CYB5R family of enzymes using an innovative “biopanning” approach. Utilizing a genetically modified soybean peroxidase coupled to CYB5R enzymes we will establish new binding partners, substrates and mechanisms for these reductases. Third, using large human data sets (i.e. n=8500 participants), we will assess if common genetic variants in CYB5R enzymes associates with diseases such as hypertension, sickle cell disease and heart failure. Mechanistic studies will determine functional impact of these variants on redox signaling pathways in cardiovascular cells. Forth, we will leverage the information gained from our studies on CYB5R3 to test if a new drug we developed called “NitroQ” and test if improves redox equilibrium and reverses hypertension, sickle cell disease and heart failure. Mechanistic studies will elucidate key targets and downstream signaling pathways influenced by NitroQ. Collectively, these fundamental basic, translational and therapeutic studies will provide a critical framework that will enable us to fill these major gaps in our knowledge and improve our understanding of redox switches in cardiovascular health and disease.