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 外周动脉疾病 (PAD) 非常普遍，并且越来越多地被认为是导致以下疾病的主要原因 心血管疾病（CVD）和公共卫生负担。严重肢体缺血（CLI）是最常见的疾病之一 先进的PAD。迄今为止，除了血管内或手术治疗外，很少有其他治疗方法可以恢复 缺血组织中的血流是可用的。因此，识别新颖的 体内治疗缺血性血管损伤的策略。转录因子-EB (TFEB) 是重要的调节因子 溶酶体生物发生和自噬。 然而，TFEB 在血管疾病中的作用仍有待进一步研究。 探索过。 一氧化氮（NO）依赖性脂肪酸硝化产物的临床检测和定量 （硝基烯烃），引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 已在人类和动物血浆中检测到纳摩尔范围内的衍生物，并显示出 对预防 CVD 的多种病理生理学方面具有深远的影响。重大进展 代谢组学和脂质组学策略将共轭亚油酸 (CLA) 确定为优先底物 用于人体脂肪酸硝化。在此，我们证明硝基共轭亚油酸（硝基-CLA）的形成 很容易转化为脉管系统中的保护机制。我们的初步数据表明硝基-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，硝基-CLA 调节 TFEB 中的自噬和管形成 依赖方式。基于这一证据，该项目将检验中心假设，即增强 硝基-CLA 的内源性产生通过促进 TFEB- 来防止缺血性血管损伤 介导的自噬导致缺血后血管生成增强。提出了两个具体目标。 目标 1：确定内皮 TFEB 对于硝基 CLA 依赖性保护性自噬至关重要，并且 体外促血管生成表型。这将通过培养中的功能获得和丧失方法来解决 硝基-CLA 处理下的原代 EC。目标 2：确定内源性硝基-CLA 可防止 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 转基因和基因敲除小鼠 以确定 TFEB 功能是硝基-CLA 促进体内血流恢复所必需的。预计 通过这个提议，我们将更好地定义 TFEB 在硝基 CLA 调节的 EC 促血管生成中的重要作用 表型和缺血后血管生成，并确定硝基-CLA作为一种新型药物的口服生物利用度 缺血性血管损伤的治疗策略。这一机理研究将为 硝基-CLA的临床应用并导致治疗或/和预防缺血性的重大突破 通过有效增加缺血后血管生成来治疗血管疾病。