Project 2: Diabetes, RAGE/DIAPH1 and Hind Limb Ischemia

项目2：糖尿病、RAGE/DIAPH1 和后肢缺血

基本信息

批准号：
10407558
负责人：
ANN MARIE SCHMIDT
金额：
$ 57.2万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10407558
关键词：
ATAC-seq Affect Amputation Atherosclerosis Binding Biological Assay Blood flow Bone Marrow Cardiovascular Diseases Cell Communication Cells Coculture Techniques Conditioned Culture Media Coronary artery Coupled Cues Cytoplasmic Tail Data Databases Diabetes Mellitus Digit structure Drug Metabolic Detoxication Endothelial Cells Excision Foundations Functional disorder Genes Genetic Transcription Goals HMGB1 Protein Homeostasis Human Incidence Inflammation Inflammatory Inflammatory Response Insulin-Dependent Diabetes Mellitus Ischemia Leukocyte L1 Antigen Complex Ligands Ligation Limb structure Link LoxP-flanked allele Mediating Microvascular Dysfunction Mitochondria Modeling Molecular Monitor Mus Myocardial Infarction Non-Insulin-Dependent Diabetes Mellitus Peripheral Peripheral arterial disease Process Property Proteins Proteomics RNA Interference Recovery Regulation Risk Factors Signal Transduction Skeletal Muscle Specificity Techniques Testing Therapeutic Time Tissues Tumor-infiltrating immune cells Vascular Diseases Vascular System Wild Type Mouse Work angiogenesis antagonist cell type diabetic effective therapy femoral artery glucose metabolism human subject in vivo irradiation ischemic injury limb ischemia lipid metabolism macrophage mitochondrial dysfunction monocyte mouse Cre recombinase new therapeutic target non-diabetic novel programs receptor for advanced glycation endproducts reconstitution recruit repaired response small molecule tissue repair transcriptome sequencing transcriptomics

项目摘要

Project Summary: Project 2 The incidence of types 1 and 2 diabetes is on the rise, which will lead to increased macro- and microvascular complications. Diabetes is a leading cause of peripheral arterial disease (PAD), a significant risk factor for amputations of digits or limbs. To date, there are no effective therapies. The ligands of the receptor for advanced glycation endproducts (RAGE), such as nonenzymatically glycated proteins (AGEs), S100/calgranulins and high mobility group box 1 (HMGB1), accumulate in non-diabetic, but especially in diabetic PAD tissues. In human subjects, RAGE and its ligands are upregulated in cardiovascular disease (CVD) and PAD tissues, in multiple cell types, but especially in monocytes/macrophages (MΦs) and endothelial cells. In murine hind limb ischemia (HLI), a model of ischemic injury to the peripheral vascular system by unilateral ligation and excision of the femoral artery (FAL), mice globally devoid of Ager (the gene encoding RAGE) display significant increases in blood flow and angiogenesis in the affected skeletal muscle in diabetes and non-diabetes vs. wild type (WT) mice. In parallel, and surprisingly, Ager deletion increased inflammatory monocyte subsets, macrophage (MΦ) content and inflammation in affected skeletal muscle. In contrast, in atherosclerotic mice and in myocardial infarct tissue (Project 1), significantly reduced MΦ tissue content and inflammation accompanied tissue repair, thereby unveiling novel niche-specific forces that regulate RAGE-dependent inflammatory responses. The cytoplasmic domain of RAGE binds to the formin, DIAPH1, which transduces RAGE ligand-stimulated signal transduction; preliminary data show that mice globally devoid of Diaph1 display significant increases in blood flow after HLI vs. WT mice. Further, our novel observation that DIAPH1 binds to Mitofusin2 (MFN2) links RAGE/DIAPH1 to mitochondrial properties and the myriad consequences for tissue homeostasis after ischemia. This Program Project shows for the first time that RAGE, DIAPH1 and MΦs co-localize in human atherosclerosis in the coronary artery. We hypothesize that RAGE/DIAPH1/MFN2-specific cues from infiltrating immune cells and/or the cellular microenvironment mediate cell-intrinsic and/or cell-cell cross-talk mechanisms in MΦs and in tissue endothelial cells (ECs) in HLI/FAL, which aggravate tissue damage and quell repair. We will employ novel Ager and Diaph1 floxed mice, small molecule antagonists of RAGE-DIAPH1 interaction and state-of-the-art molecular techniques to uncover mechanisms of diabetic PAD and to identify novel therapeutic targets and strategies. Project 2 will work closely with Projects 1 and 3 and the two Cores to achieve these goals.

项目概要：项目2 1型和2型糖尿病的发病率呈上升趋势，这将导致大血管和微血管的增加糖尿病是外周动脉疾病 (PAD) 的主要原因，而外周动脉疾病是导致外周动脉疾病的重要危险因素。迄今为止，还没有针对晚期截肢的受体的配体。糖化终产物 (RAGE)，例如非酶糖化蛋白 (AGE)、S100/钙颗粒蛋白和高迁移族蛋白 1 (HMGB1) 在非糖尿病患者的 PAD 组织中积聚，尤其是在糖尿病患者的 PAD 组织中。在多个受试者中，RAGE 及其配体在心血管疾病 (CVD) 和 PAD 组织中上调细胞类型，特别是单核细胞/巨噬细胞 (MΦs) 和内皮细胞在小鼠后肢缺血中。（HLI），通过单侧结扎和切除周围血管系统的缺血性损伤模型股动脉（FAL）中，全身缺乏 Ager（编码 RAGE 的基因）的小鼠表现出显着增加糖尿病和非糖尿病患者与野生型 (WT) 患者受影响骨骼肌中的血流和血管生成与此同时，令人惊讶的是，Ager 缺失增加了炎症单核细胞亚群、巨噬细胞 (MΦ)。相比之下，在动脉粥样硬化小鼠和心肌梗塞中，受影响的骨骼肌中的含量和炎症。组织（项目1），显着降低MΦ组织含量和伴随组织修复的炎症，从而揭示了调节 RAGE 依赖性炎症反应的新的利基特异性力量。 RAGE 结构域与福尔明 (formin)、DIAPH1 结合，可转导 RAGE 配体刺激的信号转导；初步数据显示，全身缺乏 Diaph1 的小鼠在 HLI 后血流量显着增加此外，我们的新观察发现 DIAPH1 与 Mitofusin2 (MFN2) 结合，将 RAGE/DIAPH1 与线粒体特性以及缺血后组织稳态的多种后果。项目首次表明 RAGE、DIAPH1 和 MΦ 在人类冠状动脉粥样硬化中共定位我们从浸润的免疫细胞和/或细胞中获取了 RAGE/DIAPH1/MFN2 特异性线索。微环境介导 MΦ 和组织内皮细胞中的细胞内在和/或细胞间串扰机制 HLI/FAL 中的细胞 (EC) 会加剧组织损伤并抑制修复，我们将采用新型 Ager 和 Diaph1。 floxed 小鼠、RAGE-DIAPH1 相互作用的小分子拮抗剂和最先进的分子技术项目 2 将揭示糖尿病 PAD 的机制并确定新的治疗靶点和策略。与项目 1 和 3 以及两个核心密切合作以实现这些目标。