Document text
Principal Investigator: Elena Aikawa
Organization: BRIGHAM AND WOMEN'S HOSPITAL
Fiscal Year: 2020
Award: $661,115
Funding agency: National Heart Lung and Blood Institute
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.
Terms: <3-D><3-Dimensional><3D><Acute><Affect><Antigen 60B8-A><Apo-E><ApoE><Apolipoprotein E><Area><Arterial Fatty Streak><Arterial Fatty Streaks><Atheroma><Atheromatous><Atheromatous degeneration><Atheromatous plaque><Biological Markers><Biomechanics><Blood><Blood Reticuloendothelial System><Blood Vessels><Bone Marrow Grafting><Bone Marrow Transplant><Bone Marrow Transplantation><Breast Microcalcification><Brittle Diabetes Mellitus><CABP-P14><CAGB><CAT scan><CGCB><CGLB><CT X Ray><CT Xray><CT imaging><CT scan><Calcified><Calcium-Binding Proteins><Calgranulin B><Calprotectin L1H Subunit><Cardiac infarction><Cardiovascular><Cardiovascular Body System><Cardiovascular Diseases><Cardiovascular Organ System><Cardiovascular system><Cell Body><Cells><Clinical><Computed Tomography><D-Glucose><DNA Molecular Biology><Data><Detection><Development><Dextrose><Diabetes Mellitus><Diabetic mouse><Elements><Encapsulated><Epidemic><Event><Finite Element Analyses><Finite Element Analysis><Genetic><Glucose><Heart Vascular><Human><Hydrogels><Hyperglycemia><IDDM><In Vitro><Insulin-Dependent Diabetes Mellitus><Juvenile-Onset Diabetes Mellitus><Ketosis-Prone Diabetes Mellitus><LIAG><Leanness><Leukocyte L1 Complex Heavy Chain><Leukocyte L1 Protein Heavy Chain><Link><MAC387><MRP-14 Protein><MRP14><MRP14 Protein><Macrophage Activation><Macrophage-Related Protein-14><Marrow Transplantation><Math Models><Mediating><Methods><Mice><Mice Mammals><Microcalcification><Microscopy><Migration Inhibitory Factor-Related Protein 14><Modeling><Modern Man><Molecular><Molecular Biology><Murine><Mus><Myeloid Calcium-Binding Protein p14><Myocardial Infarct><Myocardial Infarction><Network Analysis><Pathway Analysis><Patients><Phenotype><Play><Process><Proteomics><Publishing><R-Series Research Projects><R01 Mechanism><R01 Program><Research Grants><Research Project Grants><Research Project Summaries><Research Projects><Research Specimen><Resolution><Role><Rupture><S100 Calcium Binding Protein A9><S100A9><S100A9 Protein><S100A9 gene><STZ><Scanning Electron Microscopy><Short interfering RNA><Small Interfering RNA><Spatial Distribution><Specimen><Streptozocin><Streptozotocin><Stress><Sudden-Onset Diabetes Mellitus><System><T1 DM><T1 diabetes><T1D><T1DM><Testing><Thinness><Tomodensitometry><Translating><Type 1 Diabetes Mellitus><Type 1 diabetes><Type I Diabetes Mellitus><United States><Vascular calcification><X-Ray CAT Scan><X-Ray Computed Tomography><X-Ray Computerized Tomography><X-ray microtomography><Xray CAT scan><Xray Computed Tomography><Xray computerized tomography><Xray microtomography><Zanosar><atherosclerosis plaque><atherosclerotic lesions><atherosclerotic plaque><atherosclerotic plaque rupture><base><bio-markers><biologic marker><biomarker><biomechanical><calcification><cardiac infarct><cardiovascular disease risk><cardiovascular disorder><cardiovascular disorder risk><catscan><circulatory system><clinical imaging><clinical translation><computed axial tomography><computer tomography><computerized axial tomography><computerized tomography><coronary attack><coronary infarct><coronary infarction><density><developmental><diabetes><diabetes mouse model><diabetic><experience><experiment><experimental research><experimental study><extracellular vesicles><global health><heart attack><heart infarct><heart infarction><human tissue><hyperglycemic><image-based method><imaging method><imaging modality><imaging study><in vivo><innovate><innovation><innovative><insulin dependent diabetes><juvenile diabetes><juvenile diabetes mellitus><ketosis prone diabetes><lipid nanoparticle><macrophage><mathematic model><mathematical model><mathematical modeling><micro CT><micro computed tomography><microCT><microtomography><mineralization><molecular imaging><molecule imaging><mouse model><multi-modality><multidisciplinary><multimodality><murine model><nano particle><nano-sized particle><nanoparticle><nanosized particle><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><non-diabetic><nondiabetic><novel><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><osteogenic><pre-clinical><preclinical><scRNA-seq><siRNA><single cell RNA-seq><single cell RNAseq><single-cell RNA sequencing><social role><three dimensional><tool><type I diabetes><type one diabetes><vascular><vulnerable plaque>