Macrophages, Cell-Cell Communication, Ischemic Injury in Diabetes and the RAGE/DIAPH1 Signaling Axis

巨噬细胞、细胞间通讯、糖尿病缺血性损伤和 RAGE/DIAPH1 信号轴

• 批准号: 10191018
• 负责人: ANN MARIE SCHMIDT
• 金额: $ 248.62万
• 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/10191018
• 关键词: Address; Advanced Glycosylation End Products; Adverse effects; Affect; Anterior Descending Coronary Artery; Atherosclerosis; Beds; Binding; Biological Assay; Biology; Biophysics; Biostatistics Core; Blood flow; Bone Marrow Transplantation; Calcium; Cardiac Myocytes; Cardiovascular system; Cell Communication; Cell Death Signaling Process; Cell Therapy; Cells; Complex; Complications of Diabetes Mellitus; Cues; Cytoplasmic Tail; Development; Diabetes Mellitus; Diabetic mouse; Endoplasmic Reticulum; Endothelial Cells; Fluorescence; Generations; Genes; HMGB1 Protein; Heart; Hyperglycemia; Infiltration; Inflammation; Inflammation Mediators; Inflammatory Response; Injury; Insulin-Dependent Diabetes Mellitus; Ischemia; Left; Leukocyte L1 Antigen Complex; Ligands; Ligation; Limb structure; LoxP-flanked allele; Measures; Mediating; Metabolic dysfunction; Mitochondria; Modeling; Molecular; Molecular Biology Techniques; Mus; Myelogenous; Myocardial Infarction; NMR Spectroscopy; Non-Insulin-Dependent Diabetes Mellitus; Organ; Organ failure; Oxidative Stress; Oxides; Pathology; Peripheral; Peripheral arterial disease; Pharmacology; Pharmacotherapy; Physiological; Process; Property; Proteins; Proteomics; RNA Interference; Recovery; Regulation; Reperfusion Injury; Reperfusion Therapy; Signal Transduction; Site; Skeletal Muscle; Stress; Structure; System; Testing; Therapeutic; Tissues; Work; angiogenesis; base; data management; diabetic; femoral artery; in vivo; innovation; irradiation; ischemic injury; limb ischemia; macrophage; mitochondrial dysfunction; monocyte; mouse model; mutant; non-diabetic; novel; novel therapeutic intervention; novel therapeutics; programs; receptor for advanced glycation endproducts; receptor function; recruit; repaired; response; small molecule; structural biology; tissue repair; transcriptome sequencing

Project Summary: OVERALL Ischemia, a complication of diabetic cardiovascular (CVD) and peripheral arterial disease (PAD), is accompanied by the recruitment, infiltration and activation of monocytes/macrophages (MΦs), into affected tissues. In diabetes, MΦ properties are perturbed and repair is significantly mitigated, leading to organ failure. The microenvironments in the heart vs. the skeletal muscle display unique responses to ischemia, but the mechanisms are not fully understood. The ligands of the receptor for advanced glycation endproducts (RAGE), such as nonenzymatically glycated and oxidized advanced glycation endproducts; S100/calgranulins and high mobility group box 1, which accumulate in non-diabetic and diabetic CVD and PAD tissue, and RAGE itself, contribute to MΦ and niche-specific responses to ischemia. Mice globally devoid of Ager (the gene encoding RAGE) or devoid of myeloid Ager (lethal irradiation/bone marrow transplantation) are protected from the adverse effects of ligation of the left anterior descending coronary artery and the femoral artery, models for myocardial infarction (MI) and hind limb ischemia (HLI), respectively. In MI and HLI models, Ager deletion is accompanied by a marked reduction in tissue MΦ content and reduced expression of inflammatory mediators. Surprisingly, in HLI, deletion of Ager is accompanied by increased MΦ content and expression of inflammatory mediators in the skeletal muscle. Yet, in both cases, Ager deletion augured repair. Our discovery that the cytoplasmic domain of RAGE interaction with the formin, DIAPH1, mediates signal transduction, generation of oxidative stress and mitochondrial dysfunction (on account of our novel discovery that DIAPH1 binds to Mitofusin2 (MFN2) in ischemic tissue MΦs, cardiomyocytes and endothelial cells), may hold the key to these RAGE-dependent findings. The three Projects of this Program will use novel mouse models, state-of-the-art molecular biology techniques, novel small molecule antagonists of RAGE-DIAPH1 interaction, NMR spectroscopy and in-cell fluorescence assays to test the hypothesis that RAGE/DIAPH1 contributes to MΦ cell-intrinsic and MΦ- cardiomyocyte cross talk in MI and to and MΦ-endothelial cell cross talk in HLI, thereby amplifying tissue damage. We posit that RAGE-DIAPH1 and DIAPH1-MFN2 interactions control MΦ inflammation and that pharmacological blockade of RAGE-DIAPH1-MFN2 interaction and/or administration of monocytes/MΦs devoid of Ager or Diaph1 will facilitate the transition from pro-injury to adaptive MΦ inflammation and, thereby, hasten tissue repair in the diabetic heart and peripheral arterial systems. The meticulous integration of in vivo biology and molecular mechanisms studies (Projects 1 and 2) with structural biology (Project 3) assures the innovation, significance and ultimate relevance of this work for the development of novel therapeutic strategies for diabetes and ischemia.

项目概要：总体 缺血是糖尿病心血管 (CVD) 和外周动脉疾病 (PAD) 的并发症，伴随着 通过单核细胞/巨噬细胞 (MΦs) 的募集、渗透和激活进入受影响的组织。在 糖尿病时，MΦ 特性受到干扰，修复能力显着减弱，导致器官衰竭。这 心脏与骨骼肌的微环境对缺血表现出独特的反应，但 机制尚未完全了解。晚期糖基化终产物（RAGE）受体的配体， 例如非酶糖化和氧化的高级糖化终产物； S100/钙颗粒蛋白和高 移动组框 1（在非糖尿病和糖尿病 CVD 和 PAD 组织中积累）以及 RAGE 本身， 有助于 MΦ 和针对缺血的特定生态位反应。小鼠体内全面缺乏 Ager（编码基因） RAGE）或缺乏骨髓 Ager（致死性放射/骨髓移植）可免受不利影响 结扎左冠状动脉前降支和股动脉对心肌模型的影响 分别为梗塞（MI）和后肢缺血（HLI）。在 MI 和 HLI 模型中，Ager 删除伴随 通过显着减少组织 MΦ 含量和减少炎症介质的表达。令人惊讶的是，在 HLI，Ager 的缺失伴随着 MΦ 含量和炎症介质表达的增加 骨骼肌。然而，在这两种情况下，阿杰的删除都预示着修复。我们发现细胞质结构域 RAGE 与福尔明、DIAPH1 相互作用，介导信号转导、氧化应激的产生和 线粒体功能障碍（由于我们新发现 DIAPH1 与 Mitofusin2 (MFN2) 结合） 缺血组织 MΦ（心肌细胞和内皮细胞），可能是这些 RAGE 依赖性的关键 发现。该计划的三个项目将使用新型小鼠模型、最先进的分子生物学 技术、RAGE-DIAPH1 相互作用的新型小分子拮抗剂、NMR 波谱和细胞内 荧光测定来检验 RAGE/DIAPH1 有助于 MΦ 细胞固有和 MΦ- 的假设 MI 中的心肌细胞串扰以及 HLI 中的 MΦ 内皮细胞串扰，从而放大组织 损害。我们假设 RAGE-DIAPH1 和 DIAPH1-MFN2 相互作用控制 MΦ 炎症，并且 药物阻断 RAGE-DIAPH1-MFN2 相互作用和/或缺乏单核细胞/MΦ 的给药 Ager 或 Diaph1 的表达将促进从促损伤向适应性 MΦ 炎症的转变，从而加速 糖尿病心脏和外周动脉系统的组织修复。体内生物学的细致整合 分子机制研究（项目 1 和 2）与结构生物学（项目 3）确保了创新， 这项工作对于开发糖尿病新治疗策略的意义和最终相关性 和缺血。