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

基本信息

批准号：
10407554
负责人：
ANN MARIE SCHMIDT
金额：
$ 248.03万
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/10407554
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 antagonist 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

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 antagonist 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

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项目摘要

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.

项目摘要：总体缺血，糖尿病心血管（CVD）和周围动脉疾病（PAD）的并发症已完成通过募集，单核细胞/巨噬细胞（MφS）的募集，浸润和激活。糖尿病，Mφ特性受到干扰，并显着减轻修复，从而导致器官衰竭。这心脏中的微环境与骨骼肌表现出对缺血的独特反应，但机制尚未完全理解。高级糖基化最终产物（RAGE）的接收器配体，例如非酶糖化和氧化的晚期糖基化最终产物； S100/calgranulins和高流动性组框1，积累在非糖尿病和糖尿病CVD和垫组织中，愤怒本身，有助于对缺血的Mφ和小众特异性反应。全球无ager的小鼠（基因编码愤怒）或缺乏髓样脂肪（致命的辐照/骨髓移植）受到保护左前降冠状动脉和股动脉的连接的影响，心肌模型梗塞（MI）和后肢缺血（HLI）。在MI和HLI模型中，AGER缺失已完成通过明显减少组织Mφ含量和炎症介质的表达降低。令人惊讶的是，在 HLI，通过增加的Mφ含量和炎症介体在炎症中的表达来实现AGER的缺失骨骼肌。然而，在这两种情况下，Ager删除都进行了修复。我们发现的细胞质结构域与formin，Diaph1的愤怒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（由于我们的新发现，Diaph1与Mitofusin2（MFN2）结合缺血组织Mφs，心肌细胞和内皮细胞）可能会持有这些愤怒依赖性的关键发现。该程序的三个项目将使用新颖的鼠标模型，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光谱和细胞内荧光阿萨斯测试了愤怒/diaph1有助于Mφ细胞中心和Mφ-的假设。心肌细胞在MI和MI and和Mφ-内层细胞交叉中的交叉讲座，从而扩增组织损害。我们阳性rage-diaph1和diaph1-mfn2相互作用控制Mφ注入，并且 RAGE-DIAPH1-MFN2相互作用和/或单核细胞/MφS毫米的药理阻滞 Ager或Diaph1的of将有助于从亲伤害到自适应Mφ注入的过渡，从而加快糖尿病心脏和周围动脉系统的组织修复。体内生物学的细致整合与结构生物学（项目3）的分子机制研究（项目1和2）假定创新，这项工作的意义和最终相关性对于开发糖尿病的新型治疗策略和缺血。