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

项目概要：项目2 1型和2型糖尿病的发病率正在上升，这将导致大血管和微血管病变增加。 并发症糖尿病是外周动脉疾病（PAD）的主要原因，PAD是糖尿病的重要风险因素。 手指或四肢的截肢。到目前为止，还没有有效的治疗方法。晚期乳腺癌受体的配体 糖基化终产物（glycation endproducts，AGEs），如非酶促糖化蛋白（nonenzymatically glycated protein，AGEs）、S100/钙粒蛋白（calgranulin）和高糖基化蛋白（high glycation protein，HSG）。 运动族蛋白1（HMGB 1）在非糖尿病患者中积累，但尤其是在糖尿病PAD组织中。人 在受试者中，在心血管疾病（CVD）和PAD组织中， 细胞类型，但尤其是单核细胞/巨噬细胞（MΦs）和内皮细胞。在小鼠后肢缺血 (HLI)，通过单侧结扎和切除外周血管系统的缺血性损伤模型， 在股动脉（FAL）中，完全缺乏Ager（编码Ager的基因）的小鼠显示出显著增加的 糖尿病和非糖尿病与野生型（WT）受影响骨骼肌中的血流和血管生成 小鼠同时，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） 含量和炎症的影响骨骼肌。相反，在动脉粥样硬化小鼠和心肌梗死小鼠中， 组织（项目1），显著减少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。细胞质 CD 34的结构域与CD 34，DIAPH 1结合，DIAPH 1转导CD 34配体刺激的信号转导; 初步数据显示，总体上缺乏Diaph 1的小鼠在HLI后显示血流量显著增加， vs. WT小鼠。此外，我们的新观察，即DIAPH 1与线粒体融合蛋白2（MFN 2）结合， 线粒体特性和缺血后组织稳态的无数后果。这个程序 该项目首次显示，在冠状动脉粥样硬化中，CD 4，DIAPH 1和MΦs共定位于人类动脉粥样硬化中 动脉我们假设来自浸润性免疫细胞和/或细胞免疫球蛋白的CD 45/DIAPH 1/MFN 2特异性信号， 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中的内皮细胞（EC），其加重组织损伤并抑制修复。我们将聘请小说家阿格和黛芬妮 floxed小鼠，RAGE-DIAPH 1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PAD的发病机制，确定新的治疗靶点和策略。项目2将 与项目1和3以及两个核心密切合作，以实现这些目标。

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.