Transcription Factor-EB and Postischemic Angiogenesis

转录因子-EB 与缺血后血管生成

• 批准号: 9368394
• 负责人: Yanbo Fan
• 金额: $ 42.54万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9368394
• 关键词: Address; Animal Model; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Autophagocytosis; Bioavailable; Biogenesis; Biological Availability; Blood Vessels; Blood flow; Cardiovascular Diseases; Cardiovascular system; Cell Nucleus; Cell Proliferation; Cell Therapy; Clinical; Conjugated Linoleic Acids; Data; Dependency; Detection; Diet; Endothelial Cells; Endothelium; Fatty Acids; Foundations; Genetic Engineering; Genetic Transcription; Heart; Hindlimb; Human; Huntington Disease; Impairment; In Vitro; Injury; Intervention; Ischemia; Knockout Mice; Lead; Limb structure; Linoleic Acids; Lipids; Mediating; Modeling; Morbidity - disease rate; Mus; Neurodegenerative Disorders; Nitrates; Nitric Oxide; Nitrites; Oils; Operative Surgical Procedures; Oral; Outcome; Parkinson Disease; Peripheral arterial disease; Pharmacology; Phenotype; Plasma; Prevention; Production; Proteins; Public Health; Quality of life; Recovery; Regulation; Research; Risk; Role; Signal Transduction; Site; Solid; Supplementation; Testing; Therapeutic; Tissues; Transgenes; Transgenic Mice; Transgenic Organisms; Translating; Tube; Vascular Diseases; angiogenesis; base; gene therapy; in vivo; interest; loss of function; metabolomics; migration; modifiable risk; nanomolar; nitration; nitroalkene; novel; novel strategies; novel therapeutics; preclinical study; prevent; protective effect; tool; transcription factor

ABSTRACT Peripheral arterial disease (PAD) is highly prevalent and is increasingly recognized as a major contributor to the cardiovascular disease (CVD) and public health burden. Critical limb ischemia (CLI) is one of the most advanced PAD. To date, beside endovascular or surgical treatment, few therapeutic alternatives to restore the blood flow in ischemic tissues are available. Therefore, it would be of high significance to identify novel strategies to treat ischemic vascular injury in vivo. Transcription factor-EB (TFEB) is a crucial regulator of lysosomal biogenesis and autophagy. However, the functions of TFEB in vascular disease remain to be explored. The clinical detection and quantitation of nitric oxide (NO)-dependent fatty acid nitration products (nitroalkenes), has sparked the interest on novel anti-inflammatory lipids. Free and esterified nitro-fatty acid derivatives have been detected in human and animal plasma in the nanomolar range and shown to have profound implications in the prevention of diverse pathophysiological aspects of CVDs. Significant advances in metabolomics and lipidomics strategies identified conjugated linoleic acid (CLA) as the preferential substrate for fatty acid nitration in humans. Herein, we show that nitro-conjugated linoleic acid (nitro-CLA) formation readily translates into protective mechanisms in the vasculature. Our preliminary data indicate that nitro-CLA enhances autophagy through TFEB in endothelial cells (ECs). Endothelial TFEB significantly increases postischemic angiogenesis in vivo. Finally, nitro-CLA regulates autophagy and tube formation in a TFEB- dependent manner. Based on this evidence, the project will test the central hypothesis that enhancing the endogenous production of nitro-CLA protects against ischemic vascular injury by promoting TFEB- mediated autophagy leading to enhanced postischemic angiogenesis. Two Specific Aims are proposed. Aim 1: Establish that endothelial TFEB is essential for nitro-CLA-dependent protective autophagy and proangiogenic phenotype in vitro. This will be addressed by gain- and loss-of-function approaches in cultured primary ECs in the presence of nitro-CLA treatment. Aim 2: Establish that endogenous nitro-CLA protects from ischemic vascular injury in vivo through endothelial TFEB. An oral therapeutic strategy to promote endogenous nitro-CLA formation will be established. Unique EC-selective TFEB transgenic and knockout mice will be used to establish that TFEB function is required for nitro-CLA-promoting blood flow recovery in vivo. It is expected that with this proposal we will better define the essential role of TFEB in nitro-CLA-regulated EC proangiogenic phenotype and postischemic angiogenesis, and establish oral bioavailability of nitro-CLA as a novel therapeutic strategy against ischemic vascular injury. This mechanistic research will set a solid foundation for clinical utilization of nitro-CLA and lead to a major breakthrough for treating or/and preventing ischemic vascular disease by efficiently increasing postischemic angiogenesis.

抽象的 外围动脉疾病（PAD）高度普遍，越来越多地被认为是造成的主要贡献者 心血管疾病（CVD）和公共卫生负担。关键的肢体缺血（CLI）是最多的。 高级垫。迄今为止，除了血管内或外科治疗外，几乎没有治疗替代方法可以恢复 有缺血组织中的血流可用。因此，识别新颖的意义将具有很高的意义 治疗体内缺血性血管损伤的策略。转录因子-EB（TFEB）是关键的调节剂 溶酶体生物发生和自噬。 但是，TFEB在血管疾病中的功能仍然是 探索。 一氧化氮（NO）依赖性脂肪酸硝化产物的临床检测和定量 （Nitroalkenes）激发了人们对新型抗炎脂质的兴趣。游离和酯化的硝基脂肪酸 在纳摩尔范围内的人和动物血浆中检测到衍生物 在预防CVD的各种病理生理方面的深刻含义。重大进展 代谢组学和脂肪组学策略将共轭亚油酸（CLA）鉴定为优先底物 用于人类的脂肪酸硝化。在此，我们表明硝基偶联的亚油酸（Nitro-CLA）形成 容易转化为脉管系统中的保护机制。我们的初步数据表明Nitro-CLA 通过内皮细胞（EC）中的TFEB增强自噬。内皮TFEB显着增加 体内缺血后血管生成。最后，Nitro-CLA调节TFEB-中的自噬和管形成 依赖方式。基于此证据，该项目将检验中心假设，以增强 硝基-CLA的内源性产生可通过促进TFEB-可预防缺血性血管损伤 介导的自噬导致疾病后血管生成增强。提出了两个具体目标。 目标1：确定内皮TFEB对于依赖硝基-CLA的保护性自噬至关重要 体外促血管生成表型。这将通过培养的功能丧失方法来解决 在硝基-CLA处理的情况下，主要EC。目标2：确定内源性硝基-CLA可保护 通过内皮TFEB体内缺血性血管损伤。促进内源性的口服治疗策略 将建立硝基-CLA形成。将使用独特的EC选择性TFEB转基因和淘汰小鼠 为了确定在体内促进Nitro-CLA的血流恢复所必需的TFEB功能。预计 通过此提案，我们将更好地定义TFEB在硝基-CLA调节的EC中的重要作用 表型和缺血后血管生成，并建立硝基-CLA的口服生物利用度作为一种新型 针对缺血性血管损伤的治疗策略。这项机械研究将为 硝基-CLA的临床利用，导致治疗或/和预防缺血的重大突破 血管疾病通过有效增加缺血后血管生成。