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

项目2：糖尿病、RAGE/DIAPH1 和后肢缺血

• 批准号: 10642712
• 负责人: ANN MARIE SCHMIDT
• 金额: $ 57.62万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 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/10642712
• 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 gene; Homeostasis; Human; Incidence; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Insulin-Dependent Diabetes Mellitus; Ischemia; Leukocyte L1 Antigen Complex; Ligands; Ligation; Limb structure; Link; LoxP-flanked allele; Macrophage; 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; Vascular Diseases; Vascular System; Wild Type Mouse; Work; angiogenesis; antagonist; cell type; diabetic; effective therapy; femoral artery; glucose metabolism; glycation; human subject; immune cell infiltrate; in vivo; irradiation; ischemic injury; limb ischemia; lipid metabolism; 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

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 类型1和2糖尿病的事件正在上升，这将导致宏观和微血管增加 并发症。糖尿病是周围动脉疾病（PAD）的主要原因，这是一个重要的风险因素 数字或四肢的截肢。迄今为止，还没有有效的疗法。高级接收器的配体 糖基化终产（RAGE），例如非酶糖化蛋白（年龄），S100/calgranulins和High 流动性组框1（HMGB1），积聚在非糖尿病患者中，尤其是在糖尿病垫组织中。在人类中 受试者，愤怒及其配体在心血管疾病（CVD）和PAD组织中进行了更新， 细胞类型，尤其是在单核细胞/巨噬细胞（MφS）和内皮细胞中。在鼠后肢缺血中 （HLI），通过单方面结扎和切除对周围血管系统缺血性损伤的模型 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与野生型（WT） 老鼠。同时，令人惊讶的是，Ager缺失增加了炎症单核细胞亚集，巨噬细胞（Mφ） 受影响的骨骼肌的含量和感染。相反，在动脉粥样硬化小鼠和心肌保险中。 组织（项目1），显着降低了Mφ组织含量和注射完成的组织修复 揭示了调节rage依赖性炎症反应的新型小众特异性力。细胞质 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，全球没有diaph1的小鼠在HLI后的血流显着增加 vs. WT小鼠。此外，我们的新发现，即diaph1与丝曲表示（MFN2）结合，将愤怒/diaph1连接起来 线粒体特性和缺血后组织稳态的无数后果。这个程序 项目首次表明，在人动脉粥样硬化中愤怒，Diaph1和Mφs在冠状动脉粥样硬化中共定位 动脉。我们假设来自浸润的免疫细胞和/或细胞的Rage/Diaph1/MFN2特异性提示 微环境介导MφS和组织内皮中的细胞内部和/或细胞 - 细胞串扰机制 HLI/FAL中的细胞（EC），加剧组织损伤并修复平压。我们将使用新颖的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 发现糖尿病垫的机制并确定新颖的治疗靶标和策略。项目2将 与项目1和3和两个核心紧密合作以实现这些目标。