Macrophage-derived microcalcificaitons

巨噬细胞来源的微钙化

• 批准号: 9287227
• 负责人: Elena Aikawa
• 金额: $ 71.08万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-24 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9287227
• 关键词: Acute; Affect; Apolipoprotein E; Area; Arterial Fatty Streak; Biological Markers; Biomechanics; Blood; Blood Vessels; Bone Marrow Transplantation; Breast Microcalcification; Calcified; Calcium-Binding Proteins; Calgranulin B; Cardiovascular Diseases; Cardiovascular system; Cells; Clinical; Data; Detection; Development; Diabetes Mellitus; Diabetic mouse; Elements; Encapsulated; Epidemic; Event; Finite Element Analysis; Genetic; Glucose; Human; Hydrogels; Hyperglycemia; In Vitro; Insulin-Dependent Diabetes Mellitus; Link; Lipids; Macrophage Activation; Mediating; Methods; Microscopy; Modeling; Molecular; Molecular Biology; Mus; Myocardial Infarction; Pathway Analysis; Patients; Phenotype; Play; Process; Proteomics; Publishing; Research Project Grants; Research Project Summaries; Resolution; Role; Rupture; S100A9 gene; Scanning Electron Microscopy; Small Interfering RNA; Spatial Distribution; Specimen; Streptozocin; Stress; System; Testing; Thinness; Translating; United States; Vascular calcification; X-Ray Computed Tomography; base; calcification; cardiovascular disorder risk; clinical imaging; clinical translation; density; diabetic; experience; experimental study; extracellular vesicles; global health; human tissue; imaging modality; imaging study; in vivo; innovation; macrophage; mathematical model; microCT; mineralization; molecular imaging; mouse model; multidisciplinary; multimodality; nanoparticle; new therapeutic target; non-diabetic; novel; osteogenic; pre-clinical; tool; transcriptome sequencing

Project Summary This research project will test the hypothesis that, in the diabetic milieu, S100A9 induces the calcification potential of macrophage-derived extracellular vesicles (EV), precursors of microcalcifications, contributing to the biomechanical instability of the vulnerable atherosclerotic plaque. S100A9, a recently identified biomarker of vulnerable plaques, increases in the blood of patients with type 1 diabetes, is expressed by macrophages, and is a component of EV. Our published studies linked macrophages and calcification, and showed that macrophages can release EV with a high calcification potential. The present study will explore further the role of proinflammatory macrophages in vascular calcification in diabetes. Specific Aim 1 will test the hypothesis in vitro that diabetic milieu promotes macrophage activation and accelerates release and mineralization of S100A9–enriched EV. These experiments will involve innovative methods for detection of EV microcalcifications and macrophage phenotypes, including density dependent scanning electron microscopy (DDSEM) combined with elemental analysis, high-resolution microscopy, nanoparticle tracking analysis, 3D-hydrogel system, proteomics, single cell RNA sequencing, and network analyses. Specific Aim 2 will test the hypothesis in vivo that S100A9 mediates diabetes-induced microcalcifications in atherosclerotic plaques. We expect that i) genetic deletion of S100A9, ii) macrophage-targeted siRNA silencing of S100A9, and iii) bone marrow transplantation from S100A9-deficient mice will retard the progression of microcalcification and subsequent rupture, as determined by molecular imaging and comprehensive histopathological analyses. Specific Aim 3 will quantitatively evaluate the impact of microcalcification on the biomechanical instability of the atherosclerotic plaque, using mathematical modeling and finite element analysis. These complementary studies will advance the field by identifying the role of macrophage S100A9 in microcalcification. To facilitate clinical translation of mouse data, we will employ human primary macrophages and atherosclerotic plaque specimens from patients with diabetes. The findings from this project will help to develop much needed anti-calcification therapies.

项目摘要 本研究项目将验证以下假设：在糖尿病环境中，S100A9诱导钙化 巨噬细胞衍生的细胞外囊泡（EV）的潜力，微钙化的前体，有助于 易损动脉粥样硬化斑块的生物力学不稳定性。S100A9，最近发现的 易损斑块的生物标志物，1型糖尿病患者血液中的增加，由以下表达： 巨噬细胞，并且是EV的组分。我们发表的研究将巨噬细胞和钙化联系起来， 表明巨噬细胞可以释放具有高钙化潜力的EV。本研究将探讨 进一步研究促炎巨噬细胞在糖尿病血管钙化中的作用。具体目标1将 在体外测试糖尿病环境促进巨噬细胞活化并加速释放的假设， 富集S100A9的EV的矿化。这些实验将涉及创新的检测方法， EV微钙化和巨噬细胞表型，包括密度依赖性扫描电子 结合元素分析、高分辨率显微镜、纳米粒子跟踪的DDSEM 分析、3D水凝胶系统、蛋白质组学、单细胞RNA测序和网络分析。具体 目的2将在体内验证S100A9介导糖尿病诱导的微钙化的假设， 动脉粥样硬化斑块。我们预期i）S100A9的基因缺失，ii）巨噬细胞靶向siRNA S100A9的沉默，和iii）来自S100A9缺陷型小鼠的骨髓移植将延缓S100A9的表达。 微钙化的进展和随后的破裂，通过分子成像和 全面的组织病理学分析。具体目标3将定量评估 微钙化对动脉粥样硬化斑块生物力学不稳定性的影响， 建模和有限元分析。这些补充研究将通过确定 巨噬细胞S100A9在微钙化中的作用为了促进小鼠数据的临床翻译，我们将 使用人原代巨噬细胞和来自糖尿病患者的动脉粥样硬化斑块样本。 该项目的发现将有助于开发急需的抗钙化疗法。