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

摘要 我们的研究计划试图阐明氧化还原开关在化学、生物学和翻译方面的影响。 心血管系统。我们的工作揭示了细胞色素b5还原酶3(Cyb5R3)作为一种 通过将血红素和辅酶Q等底物还原到 调节活性氮和氧物种及下游信号传导。这款R35的首要目标是 研究建议是填补我们关于心血管氧化还原开关相关知识的四个主要空白 生物学。首先，我们将定义包括CyB5R1、2、4、5和他们的其他CyB5R家庭成员的角色(S) 在血管内皮细胞、平滑肌和心肌细胞中的功能。虽然在这些细胞类型中表达，但没有 定义这些酶在心血管系统中的功能的报告(S)。使用特定于新单元格类型 CyB5R1、2、4、5的敲除，我们将描述这些基因的生理和病理生理作用(S 跨内皮、平滑肌和心肌细胞的酶及其在氧化还原中的潜在作用(S) 发信号。第二，我们的目标是使用一种创新的方法为CyB5R家族酶建立新的客户蛋白 “生物传销”的方法。利用一种与CyB5R酶偶联的转基因大豆过氧化物酶，我们将 为这些还原酶建立新的结合伙伴、底物和机制。第三，使用大个子人 数据集(即n=8500名参与者)，我们将评估CYB5R酶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。机械论研究将确定 这些变异对心血管细胞氧化还原信号通路的功能影响。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”的新药是否 IF改善氧化还原平衡，逆转高血压、镰状细胞疾病和心力衰竭。机械论 研究将阐明NitroQ影响的关键靶点和下游信号通路。总而言之，这些 基础性、转化性和治疗性研究将提供一个关键框架，使我们能够 这些主要的知识差距提高了我们对氧化还原开关在心血管健康中的理解 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.