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.

项目摘要：项目2 1型和2型糖尿病的incideo正在上升，这将导致宏观和微血管增加并发症。截至截肢者或四肢糖基化终产（RAGE），例如非酶糖化蛋白（年龄），S100/calgranulins和High 迁移率组1（HMGB1），在非糖尿病中累积，尤其是在人类的糖尿病垫中受试者，愤怒和配体在心血管疾病（CVD）和PAD组织中上调，多个细胞类型，尤其是在单核细胞/巨噬细胞（MφS）和内皮细胞中。（HLI），通过单层结扎和排除的外周血管系统缺血性损伤模型股动脉（FAL），全球无ager（编码愤怒的基因）的小鼠显示出显着增加糖尿病和非糖尿病的骨骼肌的血流和血管生成小鼠平行，令人惊讶的是，敏捷的污染单核细胞群，巨噬细胞（Mφ）在受影响的骨骼肌肉中的含量和炎症。组织（项目1），显着降低了Mφ组织含量和伴随组织修复的炎症，此处揭示了依赖rage的炎症反应的小众特异性强制性愤怒的结构域与formin diaph1结合，它传递了rage配体刺激的信号转导；初步数据表明，全球无动物的小鼠在HLI后表现出明显的血液流动。与WT小鼠相比，我们的新颖观察结果是，Diaph1与Mitofusin2结合（MFN2）线粒体特性和缺血后的组织稳态的无数意见。项目表明，在人类动脉粥样硬化中愤怒，diaph1和mbycalize tirst tirst tirs tirs tirs tirs tirs the冠状动脉粥样硬化动脉。微环境介导MφS和组织内皮细胞细胞细胞细胞细胞细胞串扰机制的介导 HLI/FAL中的细胞（EC）加剧了组织损伤和平衡修复。 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揭示糖尿病垫的机制，并确定新型的Torapeatics目标和策略与项目1和3和两个核心紧密合作，以实现这些目标。