Mechanisms regulating VEGF receptors in diabetic angiogenesis

糖尿病血管生成中 VEGF 受体的调节机制

基本信息

批准号：
10219890
负责人：
Hong Chen
金额：
$ 81.45万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10219890
关键词：
Adaptor Signaling Protein Affect Amputation Angiogenesis Inhibitors Animal Model Animals Arteries Atherosclerosis Autophagocytosis Autophagosome Binding Biological Models Blood Vessels Cardiovascular Diseases Cause of Death Cell Proliferation Cell physiology Chronic Complications of Diabetes Mellitus Data Degradation Pathway Deposition Development Diabetes Mellitus Diabetic mouse Disease Endothelial Cells Endothelial Growth Factors Receptor Endothelium Family Funding Gene Expression Genetic Goals Growth Growth Factor Receptors Hyperglycemia Impairment In Vitro Infection Insulin Resistance Intention KDR gene Lead Leg Life Light Limb structure Lower Extremity Maps Mediating Medical Medical Care Costs Mission Molecular Molecular Target Mus Mutant Strains Mice Pathway interactions Patients Peripheral Peripheral Nervous System Diseases Peripheral arterial disease Phosphotransferases Play Post-Translational Regulation Process Productivity Proteins Retinal Diseases Role Signal Transduction Streptozocin Stroke Testing Therapeutic Tube Ubiquitination Ulcer United States United States National Institutes of Health Untranslated RNA Up-Regulation Vascular Endothelial Cell Vascular Endothelial Growth Factors Vascular Endothelium Vascularization activating transcription factor angiogenesis cell motility cost db/db mouse diabetic diabetic patient diabetic ulcer epigenetic regulation epsin epsin 1 experimental study forkhead protein in vitro Model in vivo in vivo evaluation limb amputation migration mouse model neovascularization new growth novel novel strategies novel therapeutic intervention novel therapeutics prevent receptor response skin ulcer targeted treatment transcription factor transcriptome sequencing ubiquitin-protein ligase wound healing

项目摘要

PROJECT SUMMARY/ABSTRACT Diabetes affects over 30 million people in the United States and costs a staggering $327 billion a year in direct medical costs and lost productivity. Diabetes adversely affects blood vessels as hyperglycemia and insulin resistance are key players in the development of atherosclerosis, peripheral neuropathy, retinopathy, and peripheral artery disease. Peripheral artery disease is a chronic condition where fatty deposits called plaques build up in the arteries to the legs, resulting in ulcerations and infections, which precedes 85% of diabetes- related amputations. Our goal is to understand how chronic hyperglycemia impairs vascular endothelial cell function to identify molecular targets that will form the basis for new therapeutic approaches to treat ulcerations and the other vascular complications of diabetes. Vascular endothelial growth factor receptors 2 and 3 (VEGFR2/3) are critical regulators of blood vessel growth or angiogenesis. These receptors are significantly reduced in the vascular endothelium of diabetic patients, resulting in inadequate angiogenesis. In our previously funded application, we showed that diabetic conditions induced expression of autophagosome proteins and promoted degradation of VEGFR2/3. In particular, we identified the protein Unc-51-like autophagy activating kinase 1 (Ulk1) as an important inhibitor of angiogenesis by stimulating autophagosome formation causing selective degradation of VEGFR2/3. Loss of endothelial Ulk1 elevated VEGFR2/3 levels and enhanced angiogenic responses such as endothelial cell proliferation, migration, and tube formation. In this competing renewal application, we present compelling preliminary data demonstrating the Forkhead box O1 transcription factor (FoxO1) controls expression of endothelial Ulk1 in both in vivo and in vitro diabetic model systems and that deficiency of endothelial FoxO1 inhibits autophagosome formation. We also show the noncoding RNA miR183-3p inhibits endothelial FoxO1 expression and the deficiency of epsin 1 and 2 adaptor proteins promotes FoxO1 ubiquitination and degradation in diabetes. These findings strongly suggest that targeting FoxO1 to protect VEGFR2/3 from degradation may represent a novel therapeutic strategy to prevent inadequate vascularization in diabetic ulcers. In view of that, we will investigate the following Specific Aims using unique mutant mice as well as in vitro models of diabetes: 1) determine the molecular mechanisms underlying FoxO1-mediated inhibition of neovascularization in diabetes, 2) determine the molecular mechanisms regulating FoxO1 activity in the diabetic endothelium, and 3) determine the therapeutic potential of targeting FoxO1 by genetic deletion or miR183-3p-mediated inhibition in diabetic angiogenesis. Our findings will enhance understanding of the cellular mechanisms behind VEGFR2/3 loss and activation of FoxO1 in regulating blood vessel damage in diabetes. We anticipate that therapies targeting FoxO1 may be useful for restoring peripheral angiogenesis to ameliorate the vascular complications associated with diabetes.

项目概要/摘要糖尿病影响美国超过 3000 万人，每年造成的直接损失高达 3270 亿美元医疗费用和生产力损失。糖尿病对血管产生不利影响，如高血糖和胰岛素抵抗力是动脉粥样硬化、周围神经病、视网膜病等发展的关键因素周围动脉疾病。外周动脉疾病是一种慢性疾病，其中脂肪沉积物称为斑块积聚在腿部动脉中，导致溃疡和感染，这先于 85% 的糖尿病—— 相关截肢。我们的目标是了解慢性高血糖如何损害血管内皮细胞识别分子靶标的功能，为治疗溃疡的新治疗方法奠定基础以及糖尿病的其他血管并发症。血管内皮生长因子受体 2 和 3 (VEGFR2/3) 是血管生长或血管生成的关键调节因子。这些受体显着糖尿病患者的血管内皮细胞减少，导致血管生成不足。在我们的在之前资助的申请中，我们发现糖尿病条件诱导自噬体的表达蛋白质并促进 VEGFR2/3 的降解。特别是，我们鉴定了类似 Unc-51 的自噬蛋白通过刺激自噬体形成激活激酶 1 (Ulk1) 作为血管生成的重要抑制剂引起 VEGFR2/3 的选择性降解。内皮 Ulk1 缺失会升高 VEGFR2/3 水平，增强血管生成反应，例如内皮细胞增殖、迁移和管形成。在这个在竞争更新应用中，我们提供了令人信服的初步数据，展示了 Forkhead Box O1 转录因子 (FoxO1) 在体内和体外糖尿病模型中控制内皮 Ulk1 的表达系统，内皮细胞 FoxO1 的缺陷会抑制自噬体的形成。我们还展示了非编码RNA miR183-3p抑制内皮FoxO1表达以及epsin 1和2接头的缺乏蛋白质促进糖尿病中 FoxO1 泛素化和降解。这些发现强烈表明靶向 FoxO1 来保护 VEGFR2/3 免遭降解可能代表一种新的治疗策略，以预防糖尿病溃疡的血管化不足。鉴于此，我们将研究以下具体目标使用独特的突变小鼠以及糖尿病体外模型：1）确定分子机制 FoxO1 介导的糖尿病新生血管形成的潜在抑制作用，2) 确定分子调节糖尿病内皮细胞中 FoxO1 活性的机制，以及 3) 确定治疗潜力通过遗传删除或 miR183-3p 介导的抑制糖尿病血管生成来靶向 FoxO1。我们的发现将增强对 VEGFR2/3 丢失和 FoxO1 激活背后细胞机制的理解调节糖尿病的血管损伤。我们预计针对 FoxO1 的疗法可能有助于恢复外周血管生成以改善与糖尿病相关的血管并发症。