INFLAMMATORY CELL SIGNALING BY CD36
Roy L Silverstein, Dept Chairman
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Grant 5P01HL087018-03_0004 from National Heart, Lung, And Blood Institute
Abstract: THE Dpnwincn Interaction of atherogenic lipids with vascular cells plays a critical role in the formation and progression of atherosclerotic lesions. The goal of this project is to define the mechanisms by which CD36, a scavenger receptor for specific forms of oxidized phospholipid, mediates pro-atherogenic and pro-inflammatory responses. Recent studies suggest that scavenger receptor function in the vessel wall may be much more complex than simply serving as a conduit for uptake of atherogenic LDL particles and that intracellular signals triggered by the interaction of oxidized lipids with macrophage CD36 may induce responses that contribute to lesion development and plaque instability. New data obtained during the previous grant period shows that CD36-mediated signals led to activation of MAP kinases JNK-1 and -2. Given the central role of JNK in mediating pro-inflammatory responses and new studies linking pathogenesis of diabetes and insulin resistance to JNK activation, we propose that a CD36-dependent signaling pathway induced by the interaction of specific oxidized phospholipids with macrophages and adipocytes contributes to atherosclerosis and provides a mechanistic connection among atherosclerosis, inflammation, and insulin resistance. To explore this hypothesis, we will take advantage of unique reagents and expertise available through this new Program Project, including well characterized oxidized phospholipids that function as specific CD36 ligands, multiple cd36 null mouse strains, and technologies to transduce primary monocytes with cDNA and RNAi constructs. The 1st aim will define the role of specific oxidized lipid ligands in activating macrophage CD36, identify the molecular elements of the CD36 signaling pathway in macrophages, characterize the intracellular signaling pathways induced by CD36, and define the mechanisms by which CD36 cross talks with other vascular cell receptor signaling pathways, focusing on receptors of the innate immune system and insulin receptor. The 2nd aim will define mechanisms by which CD36 signals regulate specific macrophage functions; e.g. lipoprotein and apoptotic cell uptake, modulation of the inflammatory response, and cell migration
Keywords: APOE [{C0003595}]; Address; Adipocytes; Adipose Cell; Apo-E; ApoE; Apolipoprotein E; Apoptotic; Arterial Fatty Streak; Atheroma; Atheromatous; Atheromatous degeneration; Atheromatous plaque; Atheroscleroses; Atherosclerosis; Atherosclerotic Cardiovascular Disease; Blood Vessels; Blood monocyte; CD36; CD36 gene; Cell Communication and Signaling; Cell Locomotion; Cell Migration; Cell Movement; Cell Signaling; Cells; Cellular Migration; Chronic; Complementary DNA; Complex; DNA, Complementary; Data; Development; Diabetes Mellitus; Disease; Disorder; EC 2.7.2-; Elements; Extracellular Signal-Regulated Kinases; Fat Cells; Foam Cells; GP3B; GP4; GPIV; Goals; Grant; Human; Human, General; INFLM; INSR; Immune system; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Insulin Receptor; Insulin Receptor Protein-Tyrosine Kinase; Insulin Resistance; Insulin-Dependent Tyrosine Protein Kinase; Intracellular Communication and Signaling; JNK; JNK1; JNK1A2; JNK21B1/2; Knockout Mice; LDL; Lesion; Ligands; Link; Lipids; Lipocytes; Lipoprotein (a); Lipoprotein Lp(a); Lipoproteins; Lipoproteins, LDL; Low-Density Lipoproteins; Lp(a); MAP Kinase 8 Gene; MAP kinase; MAPK; MAPK8; MAPK8 gene; Mammals, Mice; Man (Taxonomy); Man, Modern; Marrow monocyte; Mature Lipocyte; Mature fat cell; Mediating; Mediator; Mediator of Activation; Mediator of activation protein; Metabolic Pathway; Metabolic syndrome; Mice; Mice, Knock-out; Mice, Knockout; Mitogen-Activated Protein Kinases; Molecular; Motility; Motility, Cellular; Mouse Strains; Murine; Mus; Null Mouse; Obesity; PRKM8; Pathogenesis; Pathway interactions; Phosphatides; Phospholipids; Play; Post-Transcriptional Gene Silencing; Post-Transcriptional Gene Silencings; Posttranscriptional Gene Silencing; Posttranscriptional Gene Silencings; Process; Programs (PT); Programs [Publication Type]; Publishing; Quelling; RNA Interference; RNA Silencing; RNA Silencings; RNAi; Reagent; Receptor Protein; Receptor Signaling; Role; SAPK1; SCARB3; Sequence-Specific Posttranscriptional Gene Silencing; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Streaks, Arterial Fatty; Technology; acetylated LDL receptor; adiposity; atherogenesis; atheromatosis; atherosclerosis plaque; atherosclerotic lesions; atherosclerotic plaque; atherosclerotic vascular disease; beta-Lipoproteins; biological signal transduction; body system, allergic/immunologic; cDNA; cell motility; corpulence; corpulency; corpulentia; diabetes; disease/disorder; insulin resistant; interest; macrophage; macrophage scavenger receptors; migration; monocyte; obese; obese people; obese person; obese population; organ system, allergic/immunologic; oxidized lipid; particle; pathway; programs; receptor; receptor function; receptor, acetyl-LDL; response; scavenger receptor; scavenger receptors, macrophage; social role; uptake; vascular; vulnerable plaque
Budget start date: 1-MAY-2009
Budget end date: 30-APR-2010
5P01HL087018-03_0004 (2009): $392140
Sponsored Links Excellgen http://Excellgen.com
Roy L Silverstein, Dept Chairman
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Grant 5P01HL087018-03_9003 from National Heart, Lung, And Blood Institute
Abstract: The purpose of Core A, Administration is to (1) provide administrative and biostatistics support for the Program Director and all of the project and core investigators, and to (2) promote tight coordination, organization and integration of the four Projects and the two scientific cores to maximize productivity and leverage the interdisciplinary synergies of the program. The Administrative Core will be responsible for 1) All fiscal matters, including Preparation of monthly statements and annual budgets for the Program Director and each Project and Core Leader; Coordination of all purchase orders for equipment and service contracts; Tracking the proportionate effort of all individuals in projects and cores to assure compliance with the budget. 2.) Arrangement and scheduling of monthly meetings of the Steering Committee to discuss the program´s progress, both with respect to scientific as well as operational issues. The steering committee will consist of the Pi´s of each project and co-Pi´s of each core along. 3.) Scheduling of bi-weekly research in progress seminar series. 4.) Quality control oversight of all communications derived from Program Project activities, including manuscripts, s, oral or poster presentations at scientific and medical symposia, and press releases to translate the findings for the lay community. 5.) Compliance with all institutional and federal processes, including the tracking and oversight of all Human Subject and Animal Research Protocol submissions, and compliance with OSHA, Radiation Safety, and HIPAA regulations. 6.) Communication to all program Pi´s and Co-Pi´s via e-mail, phone and meetings all information pertinent to the program such as electronic pdf files of newly published manuscripts or summaries of presentations at regional or national meeting that relate to the program. 7.) Provide clerical support for all matters related to the program. 8.) Provide biostatistics support for planning and interpreting mouse and human studies; 9.) Arrange the Internal and External Advisory Board meetings to review the program. 10.) Arrange the invitation and visits for outside speakers for the program
Keywords: Animal Experimental Use; Animal Experimentation; Animal Research; Biometrics; Biometry; Biometry and Biostatistics; Biostatistics; Blood Vessels; Budgets; Communication; Communities; Contract Services; Electronics; Equipment; HIPAA; Health Insurance Portability and Accountability Act; Human; Human, General; Individual; Investigators; Kennedy Kassebaum Act; Mails; Mammals, Mice; Man (Taxonomy); Man, Modern; Manuscripts; Medical; Mice; Murine; Mus; Oral; PL 104-191; PL104-191; Phone; Phosphatides; Phospholipids; Posters; Posters [Publication Type]; Preparation; Press Releases; Press Releases (PT); Process; Productivity; Program Reviews; Programs (PT); Programs [Publication Type]; Protocol; Protocols documentation; Public Law 104-191; Publishing; Quality Control; Radiation; Regulation; Research; Research Personnel; Researchers; SCHED; Safety; Schedule; Series; Telephone; Translating; Translatings; United States Health Insurance Portability and Accountability Act; Visit; ing; conference; human subject; language translation; meetings; posters; programs; ray (radiation); statistics/biometry; symposium; vascular
Budget start date: 1-MAY-2009
Budget end date: 30-APR-2010
5P01HL087018-03_9003 (2009): $135903
5P01HL087018-04_9003 (0000): $135877
OXIDIZED PHOSPHOLIPIDS IN VASCULAR PATHOBIOLOGY
Roy L Silverstein, Dept Chairman
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Grant 5P01HL087018-04 from National Heart, Lung, And Blood Institute
Abstract: The goal of this Program is to develop a mechanistic understanding of the link between lipid oxidation and vascular pathology. The senior research team is a highly interactive multidisciplinary group with a broad range of shared expertise, including cell biology, chemical biology, biochemistry, and immunology. The 4 interrelated projects will test the hypothesis that oxidation of phospholipids in lipoproteins and cell membranes creates specific ligands for vascular cell receptors, transporters, and enzymes that then drastically alter cellular function and contribute to the pathogenesis of inflammatory diseases, including atherosclerosis. Project 1 will combine biophysical and clinical studies to define the chemical structures, biophysical properties, and oxidative pathways that participate in the formation, decay, and receptor binding activities of a novel family of specific oxidized phospholipid ligands of the Class B scavenger receptor CD36. This project will also assess the clinical relevance of specific oxidized phospholipids in human athero-sclerosis. Project 2 will determine how vascular cells clear biologically active and toxic components of oxLDL by defining the fate of extracellular oxidized phospholipids, characterizing transporters and transport mechanisms of oxidized phospholipids, and determining signaling pathways connecting oxLDL and phospholipids to cell viability. Project 3 will define complex mechanisms that regulate expression and activity of monocyte/macrophage 15- lipoxygenase, the enzyme responsible for producing 13-HPODE, an oxidation product of linoleic acid and a prominent oxidized lipid found in atheroma. Project 4 will define vascular cell signaling pathways initiated by binding of specific oxidized phospholipids to CD36, focusing on how this signaling cascade modulates foam cell formation, inflammatory responses, and metabolic pathways. An Administrative Core and 2 scientific cores, "Analytic and Synthetic Chemistry" and "Monocyte/Macrophage Cell Biology" strengthen the Program by providing well characterized, uniform-quality cellular, animal, and chemical reagents
Project start date: 2007-08-06
Project end date: 2012-04-30
Budget start date: 1-MAY-2010
Budget end date: 30-APR-2011
5P01HL087018-04 (2010): $2281238
5P01HL087018-03 (2009): $2281679
INFLAMMATORY CELL SIGNALING BY CD36
Roy L Silverstein, Dept Chairman
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Abstract: THE Dpnwincn Interaction of atherogenic lipids with vascular cells plays a critical role in the formation and progression of atherosclerotic lesions. The goal of this project is to define the mechanisms by which CD36, a scavenger receptor for specific forms of oxidized phospholipid, mediates pro-atherogenic and pro-inflammatory responses. Recent studies suggest that scavenger receptor function in the vessel wall may be much more complex than simply serving as a conduit for uptake of atherogenic LDL particles and that intracellular signals triggered by the interaction of oxidized lipids with macrophage CD36 may induce responses that contribute to lesion development and plaque instability. New data obtained during the previous grant period shows that CD36-mediated signals led to activation of MAP kinases JNK-1 and -2. Given the central role of JNK in mediating pro-inflammatory responses and new studies linking pathogenesis of diabetes and insulin resistance to JNK activation, we propose that a CD36-dependent signaling pathway induced by the interaction of specific oxidized phospholipids with macrophages and adipocytes contributes to atherosclerosis and provides a mechanistic connection among atherosclerosis, inflammation, and insulin resistance. To explore this hypothesis, we will take advantage of unique reagents and expertise available through this new Program Project, including well characterized oxidized phospholipids that function as specific CD36 ligands, multiple cd36 null mouse strains, and technologies to transduce primary monocytes with cDNA and RNAi constructs. The 1st aim will define the role of specific oxidized lipid ligands in activating macrophage CD36, identify the molecular elements of the CD36 signaling pathway in macrophages, characterize the intracellular signaling pathways induced by CD36, and define the mechanisms by which CD36 cross talks with other vascular cell receptor signaling pathways, focusing on receptors of the innate immune system and insulin receptor. The 2nd aim will define mechanisms by which CD36 signals regulate specific macrophage functions; e.g. lipoprotein and apoptotic cell uptake, modulation of the inflammatory response, and cell migration
Keywords: APOE [{C0003595}]; Address; Adipocytes; Adipose Cell; Apo-E; ApoE; Apolipoprotein E; Apoptotic; Arterial Fatty Streak; Atheroma; Atheromatous; Atheromatous degeneration; Atheromatous plaque; Atheroscleroses; Atherosclerosis; Atherosclerotic Cardiovascular Disease; Blood Vessels; Blood monocyte; CD36; CD36 gene; Cell Communication and Signaling; Cell Locomotion; Cell Migration; Cell Movement; Cell Signaling; Cells; Cellular Migration; Chronic; Complementary DNA; Complex; DNA, Complementary; Data; Development; Diabetes Mellitus; Disease; Disorder; EC 2.7.2-; Elements; Extracellular Signal-Regulated Kinases; Fat Cells; Foam Cells; GP3B; GP4; GPIV; Goals; Grant; Human; Human, General; INFLM; INSR; Immune system; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Insulin Receptor; Insulin Receptor Protein-Tyrosine Kinase; Insulin Resistance; Insulin-Dependent Tyrosine Protein Kinase; Intracellular Communication and Signaling; JNK; JNK1; JNK1A2; JNK21B1/2; Knockout Mice; LDL; Lesion; Ligands; Link; Lipids; Lipocytes; Lipoprotein (a); Lipoprotein Lp(a); Lipoproteins; Lipoproteins, LDL; Low-Density Lipoproteins; Lp(a); MAP Kinase 8 Gene; MAP kinase; MAPK; MAPK8; MAPK8 gene; Mammals, Mice; Man (Taxonomy); Man, Modern; Marrow monocyte; Mature Lipocyte; Mature fat cell; Mediating; Mediator; Mediator of Activation; Mediator of activation protein; Metabolic Pathway; Metabolic syndrome; Mice; Mice, Knock-out; Mice, Knockout; Mitogen-Activated Protein Kinases; Molecular; Motility; Motility, Cellular; Mouse Strains; Murine; Mus; Null Mouse; Obesity; PRKM8; Pathogenesis; Pathway interactions; Phosphatides; Phospholipids; Play; Post-Transcriptional Gene Silencing; Post-Transcriptional Gene Silencings; Posttranscriptional Gene Silencing; Posttranscriptional Gene Silencings; Process; Programs (PT); Programs [Publication Type]; Publishing; Quelling; RNA Interference; RNA Silencing; RNA Silencings; RNAi; Reagent; Receptor Protein; Receptor Signaling; Role; SAPK1; SCARB3; Sequence-Specific Posttranscriptional Gene Silencing; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Streaks, Arterial Fatty; Technology; acetylated LDL receptor; adiposity; atherogenesis; atheromatosis; atherosclerosis plaque; atherosclerotic lesions; atherosclerotic plaque; atherosclerotic vascular disease; beta-Lipoproteins; biological signal transduction; body system, allergic/immunologic; cDNA; cell motility; corpulence; corpulency; corpulentia; diabetes; disease/disorder; insulin resistant; interest; macrophage; macrophage scavenger receptors; migration; monocyte; obese; obese people; obese person; obese population; organ system, allergic/immunologic; oxidized lipid; particle; pathway; programs; receptor; receptor function; receptor, acetyl-LDL; response; scavenger receptor; social role; uptake; vascular; vulnerable plaque
Budget start date: 1-MAY-2010
Budget end date: 30-APR-2011
5P01HL087018-04_0004 (0000): $392065
Grants awarded to Roy L Silverstein
CD36 /TSP-1: Novel Antiangiogenic Switch Mechanism
Roy L Silverstein, Chairman
Weill Medical College Of Cornell Univ 1300 York Avenue New York, Ny 10021
Grant 5P01HL067839-050002 from National Heart, Lung, And Blood Institute IRG: ZHL1
Abstract: Thrombospondin-1 (TSP-1) is a potent inhibitor of angiogenesis in vivo and of microvascular endothelial cell (MVEC) responses to angiogenic factors in vitro. The anti-angiogeneic activity of TSP-1 is contained in a structural domain known as the type 1 repeat or TSR-1. CD36 is the cellular receptor for TSP-1 on MV3EC and is necessary for its anti- angiogenic activity. Structure-function analyses have determined that binding of TSP-1 to CD36 is mediated by interaction of the TSR-1 domain of TSP with a conserved domain called CLESH-1 in CD36. The long-term goal of this project is to characterize the anti-angiogenic switch mediated by the CD36/TSR-1 system. One specific aim is to characterize the regulation of this switch. Studies will test the hypothesis that histidine-rich glycoprotein (HRGP), a soluble CLESH-1 domain containing plasma protein, modulates the TSR-1/CD36 anti-angiogenic response in vivo. The mechanisms that control expression of CD36 on MVEC will also be studied. Other studies will determine the role of CD36 in mediating the anti-angiogenic activities of TSR-1 containing proteins other than TSP-1, such as ADAMS-TS1 and TSP-2. These studies will include determination of the ability of TSR-1 containing proteins to bind CD36 and to effect endothelial cell function in vitro and in vivo. Another aim will determine the intracellular signaling mechanisms responsible for the CD36-mediated endothelial angiostatic response. These studies will take advantage of several unique reagents, including the CD36 null mouse strain generated in our lab and peptides derived from the cytoplasmic domain of CD36. Specific experiments will identify candidate signaling molecules and pathways by characterizing proteins that interact physically with CD36 in endothelial cells. Once candidate signaling partners are identified, their role in angiogenesis will be studied using in vitro assays of endothelial function and in vivo assays of angiogenesis. The last aim will use a murine model of wound angiogenesis and a murine endothelial progenitor cell transplant model (developed by Dr. Rafii; project 4) to study the role of the CD36/TSR-1 switch in vivo. Characterization of this novel anti-angiogenic system will increase our understanding of angiogenesis and will help define potential new therapeutic targets for blood and vascular diseases and cancer. This project involves extensive use of the program cores and collaborations with other investigators in the program project, including Dr. Rafii (Project 4) to study regulation of CD36 expression in EC progenitors, and Dr. Gershengorn (Project 3) to study the impact of CD36 signaling on endothelial ell G-protein coupled receptors.
Keywords: angiogenesis, angiogenesis inhibitor, protein structure function, receptor binding, thrombospondin, vascular endothelium, CD antigen, glycoprotein, microcirculation, protein protein interaction, wound healing, laboratory mouse, stem cell transplantation
MOLECULAR AND CELL BIOLOGY OF A THROMBOSPONDIN RECEPTOR
Roy L Silverstein, Chairman
Weill Medical College Of Cornell Univ
1300 York Avenue
new York, Ny 10021
Grant 5R01HL042540-05 from National Heart, Lung, And Blood Institute IRG: HEM
Abstract: The overall objective of the proposed project is to understand the role of thrombospondin (TSP) and its cellular receptor (GPIV) in vascular biology. TSP is a major component of platelets and the vascular subendothelium and is involved in mediating platelet- platelet, platelet-monocyte and perhaps other cell-cell and cell- matrix interactions. An 88Kd integral membrane protein has recently been identified as a TSP receptor on platelets and 2 tumor ell lines. This protein is identical to platelet glycoprotein IV (GPIV) and probably to a monocyte protein of unknown function now known as CD36. TSP also modulates arterial smooth muscle cell growth and plays a role in the regulation of plasminogen activation and heparin function. Thus, study of TSP and its receptor is important in understanding both physiological and pathological events at the vessel wall (e.g. thrombosis and atherosclerosis). The specific objectives are to characterize structure-function relationships of the TSP-GPIV interaction by isolating, characterizing and cloning the cDNA for the TSP receptor (GPIV) from human endothelial cell and U937 cell cDNA libraries. The latter is a human cell line of monocyte lineage. Structural domains of GPIV, in particular those involved in mediating TSP binding, membrane insertion and intracytoplasmic functions (e.g. cytoskeletal interactions) will be characterized by analyzing function of mutated (truncated) receptor molecules in transfected cells. In a complementary manner the TSP domain involved in mediating GPIV binding will also be identified using monoclonal antibodies and partial protease digestion of the intact molecule. The gene for GPIV will be identified from a human genomic lambda phage library and partially characterized. The cellular biology of the TSP-GPIV interaction will be studied with particular reference to known TSP functions. The role of GPIV in mediating TSP-dependent cytoadhesive interactions, such as that among platelets, monocytes, and endothelia, between malaria infected erythrocytes and endothelia, and between tumor cells and extracellular matrix will be studied as will the expression and regulation of both TSP and GPIV in vascular cells (endothelium and smooth muscle) and mononuclear phagocytes using specific immunological (monoclonal and polyclonal) and molecular probes
Keywords: cell adhesion, receptor, receptor binding, thrombospondin blood vessel, cell cell interaction, monocyte, nucleic acid sequence, platelet, protein structure function, vascular endothelium, vascular smooth muscle genetic library, laboratory mouse, laboratory rabbit, molecular cloning, tissue /cell culture, transfection
Project start date: 1989-07-01
Project end date: 1994-06-30
5R01HL042540-05 (1993): $211805
5R01HL042540-04 (1992): $191553
Vascular Signaling Via Scavenger Receptors
Roy L Silverstein, Chairman
Weill Medical College Of Cornell Univ 1300 York Avenue New York, Ny 10021
Grant 5P01HL046403-140003 from National Heart, Lung, And Blood Institute
Abstract: The overall goal of this project is to characterize the molecular mechanisms of vascular cell signaling by the Class B scavenger receptor CD36. CD36 is expressed on macrophages and capillary endothelial cells, where it has been implicated in important vascular processes. As a macrophage receptor for oxidatively modified low density lipoprotein particles it is involved in directly internalization of bioactive lipids and foam cell formation during atherogenesis. As a receptor on phagocytes for apoptotic cells, it participates in their clearance and in mediating the accompanying cellular response. As a receptor on endothelium for the matrix glycoprotein thrombospondin-1 it mediates a potent anti- angiogenic activity. The molecular mechanisms by which CD36 mediates diverse actions in different cell types remain largely unknown and is the focus of this application. Dissecting these signaling pathways will have important implications in understanding regulation of macrophage and endothelial cell activation, and in defining potential new therapeutic targets for atherosclerosis inflammation, and cancer. These studies will take advantage of several unique reagents, including cells isolated from CD36 null mice, peptides derived from the cytoplasmic domain of CD36, and CD36 homolog is identified in the nematode Caenorhabditis elegans. Specific plans are to identify candidate signaling molecules and pathways involved in CD36- mediated cellular events by characterizing proteins that interact physically with CD36 in vascular cells. Strategies will include affinity chromatography on CD36 cytoplasmic peptides, co- immunoprecipitation, and yeast 2-hybrid screens. The phenotype induced by loss of expression of CD36 homologs in C. elegans will be characterized to identify the signaling pathways utilized by the homologs in engulfment of apoptotic cells and cell migration. Once candidate signaling partners are identified, their role in CD36-mediated signal transduction will be studied in vascular cells. Practical attention will be paid to changes in protein activity and localization induced by ligation of CD36 by its ligands and to their effects on angiogenesis and macrophage function. This project involves many collaborations and synergisms with investigators in other projects within the Program. Strategies to identify candidate signaling proteins by peptide sequencing will involve collaboration with Drs. Gross, K. Kajjar, and R. Upmacis (projects 5, 1 and 6), who are experts in MS/MS and protein chemistry. Studies of cytoskeletal regulation and signaling by Crk, DOCK180 and Rac will involve collaboration with Dr. Hempstead (project 4), and studies of G- protein signaling will involve collaboration with Dr. D. Hajjar. Studies of endothelial cell responses will involve collaboration with Drs. Kraemer and Hempstead (project 4).
Keywords: CD antigen, biological signal transduction, cell adhesion, cell adhesion molecule, receptor binding, receptor expression, vascular endothelium, gene expression, leukocyte activation /transformation, protein localization, protein protein interaction, Caenorhabditis elegans, affinity chromatography, enzyme linked immunosorbent assay, human subject, immunoprecipitation, laboratory mouse, protein binding, tissue /cell culture, yeast two hybrid system
REGULATION OF THE ANTI-ANTIANGIOGENIC SWITCH BY CD36, THROMBOSPONDIN, AND HRGP
Roy L Silverstein, Dept Chairman
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Grant 5R01HL085718-03 from National Heart, Lung, And Blood Institute
Abstract: The goal of this proposal is to understand the mechanisms by which angiogenesis (new blood vessel growth) is regulated by a protein receptor called CD36 that is expressed on the surface of microvascular endothelial cells. These are the cells that form the lining of blood capillaries. Angiogenesis is a critically important process in the development of many human diseases, including heart attack stroke, diabetes and cancer. Therefore, understanding how the body normally turns angiogenesis on and off has direct potential to lead to new therapeutic and prognostic approaches to many diseases. CD36 functions by sending a signal to microvascular endothelial cells to halt angiogensesis when it is exposed to any of a group of other proteins that contain a structural domain called the thrombospondin type I repeat (TSR). Our laboratory has recently disovered that a protein circulating in blood, called HRGP, bears structural similarity to CD36 and can act as a "decoy", blocking the activity of TSR-containing anti-angiogenic proteins. These discoveries have led to the hypothesis that angiogenesis is modulated by the fine control of CD36, TSR and HRGP expression in tissues. To address the hypothesis three specific aims have been developed. The first is to identify the structural determinants involved in recognition of TSR domains by CD36 and to characterize modulation of TSR binding by oxidized phospholipids. The approach will involve generating recombinant peptides, studying their interaction, and using NMR spectroscopy to define the mechanisms of the interactions at the atomic level. Aim 2 will characterize expression of CD36 in human microvascular endothelial cells, focusing on the mechanisms by which CD36 expression or function is down-regulated by modified phospholipids via protein kinase C activation and by ecto-domain phosphorylation. Expression will be measured at the mRNA and protein levels. The biological importance of these regulatory pathways in vivo will be determined in aim 3, in which angiogenesis associated with UVB skin radiation and tumor growth will be studied in mice with targeted deletion of the cd36 and hrgp genes and transgenic mice that over-express HRGP in skin. Accomplishing these aims could lead to new therapeutic approaches to modulate angiogenesis through the CD36-mediated anti-angiogenic switch. Angiogenesis (new blood vessel growth) is a critically important process in the development of many human diseases, including heart attack, stroke, diabetic retinopathy, and cancer. Therefore, understanding how the body normally turns angiogenesis on and off has direct potential to lead to new therapeutic and prognostic approaches to many diseases. This project seeks to understand the cellular mechanisms by which an anti-angiogenic switch on blood vessel lining cells (endothelial cells) is turned on and off. The switch is mediated by a cellular receptor called CD36 which functions by sending a signal to endothelial cells to halt angiogenesis when it is exposed to any of a group of other proteins that contain a structural domain called the thrombospondin type I repeat (TSR). This project will define the structural basis of TSR- CD36 interactions, determine how expression of the key components of the system are regulated, and determine the functional role of the system in regulating angiogenesis during inflammation and tumor growth in vivo
Keywords: 1-acyl-sn-glycerol-3-phosphate; 1-oleoyl-lysophosphatidic acid; Address; Angiogenesis Antagonists; Angiogenesis Blockers; Angiogenesis Inhibitors; Angiogenetic Antagonists; Angiogenic Antagonists; Angiogenic Factor; Angiogenic Proteins; Angiogenic Switch; Angiostatic Agents; Anti-Angiogenetic Agents; Anti-Angiogenic Agents; Anti-Angiogenic Drugs; Antiangiogenesis Agents; Antiangiogenic Agents; Apoplexy; Bears; Binding; Binding (Molecular Function); Biological; Blood; Blood Vessels; Blood capillaries; Body Tissues; CD36; CD36 gene; Calcium Phospholipid-Dependent Protein Kinase; Calcium-Activated Phospholipid-Dependent Kinase; Cancers; Capillaries; Cardiac infarction; Cell Communication and Signaling; Cell Line; Cell Lines, Strains; Cell Signaling; CellLine; Cells; Cerebral Stroke; Cerebrovascular Apoplexy; Cerebrovascular Stroke; Cerebrovascular accident; Development; Diabetes Mellitus; Diabetic Retinopathy; Disease; Disorder; Down-Regulation; Down-Regulation (Physiology); Downregulation; Endothelial Cells; Factor, Angiogenesis; GP3B; GP4; GPIV; Generalized Growth; Genes; Goals; Growth; HPRG; Human; Human, General; INFLM; In Vitro; Inflammation; Inhibitors, Angiogenetic; Inhibitors, Angiogenic; Intracellular Communication and Signaling; LPA; Laboratories; Lead; Ligands; Lysophosphatidic Acids; Lysophospholipids; MOPA; Malignant Neoplasms; Malignant Tumor; Mammals, Mice; Man (Taxonomy); Man, Modern; Measures; Mediating; Messenger RNA; Mice; Molecular Interaction; Murine; Mus; Myocardial Infarct; Myocardial Infarction; NMR Spectroscopy; Neovascularization Inhibitors; Organism; PKC; Pb element; Phosphatides; Phospholipid-Sensitive Calcium-Dependent Protein Kinase; Phospholipids; Phosphorylation; Post-Translational Modifications; Post-Translational Protein Processing; Posttranslational Modifications; Process; Protein Kinase C; Protein Modification; Protein Modification, Post-Translational; Protein Phosphorylation; Protein Processing, Post-Translational; Protein Processing, Posttranslational; Protein/Amino Acid Biochemistry, Post-Translational Modification; Proteins; RNA, Messenger; Radiation; Receptor Protein; Regulation; Regulatory Pathway; Reticuloendothelial System, Blood; Role; SCARB3; Signal Transduction; Signal Transduction Systems; Signaling; Skin; Spectroscopy, NMR; Stroke; Surface; System; System, LOINC Axis 4; THBS1; TSP; TSP-1; TSP1; Thrombospondin 1; Thrombospondins; Tissue Growth; Tissues; Transgenic Mice; Transgenic Organisms; UVB; UVB radiation; Ultraviolet B Radiation; Ursidae; Ursidae Family; Vascular Accident, Brain; angiogenesis; antiangiogenic; base; biological signal transduction; brain attack; cardiac infarct; cerebral vascular accident; coronary attack; coronary infarct; coronary infarction; cultured cell line; design; designing; diabetes; disease/disorder; gene product; heart attack; heart infarct; heart infarction; heavy metal Pb; heavy metal lead; histidine-proline-rich glycoprotein; histidine-rich glycoprotein; human disease; in vivo; living system; lysophosphatidic acid; mRNA; malignancy; monooleylphosphatidate; monooleylphosphatidic acid; neoplasm/cancer; new therapeutics; next generation therapeutics; novel therapeutic intervention; novel therapeutics; nuclear magnetic resonance spectroscopy; ontogeny; overexpression; prognostic; public health relevance; ray (radiation); receptor; recombinant peptide; response; social role; stroke; transgenic; tumor growth; vascular
Project start date: 2008-06-20
Project end date: 2013-05-31
Budget start date: 1-JUN-2010
Budget end date: 31-MAY-2011
PFA/PA: PA-07-070
5R01HL085718-03 (2010): $392500
Sponsored Links Excellgen http://Excellgen.com
5R01HL085718-02 (2009): $392500
Oxidized Phospholipids In Vascular Pathobiology
Roy L Silverstein, Chairman
Cleveland Clinic Lerner Col/med-cwru P84 Cleveland, Oh 44195
Grant 1P01HL087018-01A1 from National Heart, Lung, And Blood Institute IRG: HLBP
Abstract: The goal of this Program is to develop a mechanistic understanding of the link between lipid oxidation and vascular pathology. The senior research team is a highly interactive multidisciplinary group with a broad range of shared expertise, including cell biology, chemical biology, biochemistry, and immunology. The 4 interrelated projects will test the hypothesis that oxidation of phospholipids in lipoproteins and cell membranes creates specific ligands for vascular cell receptors, transporters, and enzymes that then drastically alter cellular function and contribute to the pathogenesis of inflammatory diseases, including atherosclerosis. Project 1 will combine biophysical and clinical studies to define the chemical structures, biophysical properties, and oxidative pathways that participate in the formation, decay, and receptor binding activities of a novel family of specific oxidized phospholipid ligands of the Class B scavenger receptor CD36. This project will also assess the clinical relevance of specific oxidized phospholipids in human athero-sclerosis. Project 2 will determine how vascular cells clear biologically active and toxic components of oxLDL by defining the fate of extracellular oxidized phospholipids, characterizing transporters and transport mechanisms of oxidized phospholipids, and determining signaling pathways connecting oxLDL and phospholipids to cell viability. Project 3 will define complex mechanisms that regulate expression and activity of monocyte/macrophage 15- lipoxygenase, the enzyme responsible for producing 13-HPODE, an oxidation product of linoleic acid and a prominent oxidized lipid found in atheroma. Project 4 will define vascular cell signaling pathways initiated by binding of specific oxidized phospholipids to CD36, focusing on how this signaling cascade modulates foam cell formation, inflammatory responses, and metabolic pathways. An Administrative Core and 2 scientific cores, "Analytic and Synthetic Chemistry" and "Monocyte/Macrophage Cell Biology" strengthen the Program by providing well characterized, uniform-quality cellular, animal, and chemical reagents.
Keywords: atherosclerosis, cellular pathology, inflammation, oxidized lipid, vascular endothelium, clinical research
Project start date: 2007-08-06
Project end date: 2012-04-30
1P01HL087018-01A1 (2007): $2323399
ANTIPHOSPHOLIPID ANTIBODIES IN SYSTEMIC LUPUS ERYTHEMATOSUS
Roy L Silverstein
Hospital For Special Surgery New York, Ny 10021
Grant 5P50AR042588-050003 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases
Abstract: Patients with Systemic Lupus Erythematosus often have circulating autoantibodies reactive with anionic phospholipid (aPL). The presence of these antibodies correlates with a clinical syndrome of arterial and venous thrombosis, recurrent fetal loss due to placental infarction, thrombocytopenia, and symptoms of cerebral ischemia. The pathogenesis of this hypercoagulable state is not known. The goal of this project is to elucidate the molecular basis of the prothrombotic state associated with aPL and to define genetic and other risk factors that determine risk of thrombosis in patients with aPL. Preliminary data has been obtained showing that lg purified from patients with aPL activate vascular endothelial cells (EC). This was associated with a change in the surface phenotype of the EC such that the cells became adhesive for monocytes. Treatment of EC with lg from control patients did not produce this effect, while rabbit serum from an animal immunized with apoH, a protein cofactor that makes up part of the aPL antigen, mimicked the effect. Plans are to characterize the prothrombotic phenotype induced in EC by aPL and rabbit apoH antibodies, including assays for procoagulant, anti- fibrinolytic and adhesive functions. These assays will be immunological, functional, and molecular. In addition the molecular basis of signal transduction by these antibodies will be explored, with emphasis on the role of the apoH cofactor and potential endogenous EC apoH "receptors". These receptors will be identified, characterized and molecularly cloned. We also propose to study the effect of EC adhesion on monocyte activation, since these cells are known to be important regulators of hemostasis. Since our data suggest that activated Ec are opsonized by aPL we will also explore the role of monocyte Fc receptors in contributing to the prothrombotic state. Engagement of these receptors is known to induce monocyte procoagulant activity. Additionally, activated platelets and apoptotic lymphocytes to which aPL also bind will be used as substrates for FcR-driven monocyte procoagulant activity. To bring these studies to a direct clinical level we will study a large number of patients with aPL. Since our data suggest that apoH and FcR mau play an important role in mediating the prothrombotic state associated with aPL, we will study the role of FcR genetic polymorphisms, apoH isoform, and IgG subclass in determining risk of clinical thrombosis. This will involve a multivariate analysis of a large cohort of SLE patients. In parallel, the role of these inherited factors in modulating the in vitro procoagulant state will be studied. The ultimate goal of these studies is to understand the molecular basis of the aPL syndrome and thus to develop novel diagnostic and therapeutic approaches of these patients.
Keywords: antiantibody, cell cell interaction, medical complication, molecular pathology, phospholipid, systemic lupus erythematosus, thrombosis, antibody receptor, biological signal transduction, disease /disorder proneness /risk, family genetics, genetic polymorphism, immunoglobulin G, monocyte, protein isoform, receptor expression, vascular endothelium, SDS polyacrylamide gel electrophoresis, flow cytometry, human subject, molecular cloning, northern blotting, nucleic acid probe, nucleic acid sequence, polymerase chain reaction
MOLECULAR MECHANISMS OF RPE PHAGOCYTOSIS--ROLE OF CD36
Roy L Silverstein, Chairman
Medicineweill Medical College Of Cornell Univ
1300 York Avenue
new York, Ny 10021
Grant 5R01EY010967-07 from National Eye Institute IRG: VISC
Abstract: Normal retinal function requires constant replacement of the membranous disks which comprise the outer segments of the photoreceptor cells. During this process disk membranes at the apical ends of the segments detach in packets which are ultimately phagocytosed by the retinal pigment epithelium (RPE). Dysfunction in outer segment phagocytosis by the RPE leads to retinal degeneration and blindness in experimental animals and may be an important contributor to retinal degenerative diseases in humans. The overall objective of this proposal is to characterize the molecular mechanisms by which retinal pigment epithelium recognize and phagocytose effete photoreceptor outer segments. Major emphasis will be on CD36, an 88,000MW glycoprotein that we have shown in preliminary studies to be expressed on RPE in a developmentally regulated manner, and to function as a receptor for rod outer segments (ROS). Specific aims are to use immunohistochemical, immunocytochemical, western blot, and rt-PCR to demonstrate that human RPE express CD36 protein and mRNA. Monoclonal anti- CD36 IgG, soluble cD36, and CD36 cDNA transfected cells will be used to demonstrate that CD36 is a receptor for ROS on RPE. Recombinant CD36/GST fusion proteins will be used to identify the ligand binding domain on CD36 for ROS. The ligand for CD36 on ROS will also be identified, as will other receptors that might be associated with CD36 in this function; e.g. alpha(vn)beta(3) integrin, type I/II scavenger receptors, or advanced glycation end product (AGE)-receptors. The mechanism of CD36-mediated ROS internalization will also be explored. Approaches will include electron microscopy to define the ultrastructural pathway of CD36-mediated internalization. Immunological and mutagenesis approaches will be used to define the role of caveolae and CD36-caveolin interactions in internalization. Similarly the role of CD36 interactions with cytoskeletal elements and non-receptor protein tyrosine kinases in internalization will be studied. The RCS rat is a well described animal model of retinal degeneration caused by defective RPE phagocytosis of ROS. Studies will be done to determine if the ROS uptake and internalization defect in the RCS rat is the result of an abnormality in the CD36 pathway. CD36 expression and molecular form in the RCS rat RPE will be studied as will CD36- mediated signalling pathways. Understanding the molecular details of CD36 function on RPE will provide insight into normal retinal processes such as RPE polarity and RPE phagocytosis, as well as into retinal degenerative states. Novel strategies to diagnose and treat retinal degeneration may thus result from this work
Keywords: CD antigen, phagocytosis, protein structure function, retina degeneration, retinal pigment epithelium, rod cell antigen receptor, molecular biology, protein tyrosine kinase animal tissue, electron microscopy, electroretinography, human tissue, immunocytochemistry, immunoprecipitation, laboratory mouse, laboratory rat, northern blotting, polymerase chain reaction, tissue /cell culture, transmission electron microscopy, western blotting
Project start date: 1995-05-01
Project end date: 2003-06-30
5R01EY010967-07 (2001): $275481
5R01EY010967-06 (2000): $267458
2R01EY010967-05 (1999): $259668
5R01EY010967-04 (1998): $252184
5R01EY010967-03 (1997): $242483
5R01EY010967-08 (2002): $283746
Sponsored Links Excellgen http://Excellgen.com
CELLULAR ATTACHMENT AND ADHESIVE RECEPTORS
Roy L Silverstein, Chairman
Institution:
Grant 2P01HL046403-060003 from National Heart, Lung, And Blood Institute
Abstract: Monocyte attachment, adhesion, and infiltration into the vessel wall represent hallmarks of a wide range of vascular disorders including thrombosis, atherosclerosis, and inflammation. Endothelial cell surface expression of attachment molecules, such as E- and P-selectin, and adhesion molecules, such as ICAM-1 and VCAM-1, locally induced by inflammatory mediators, plays an important role in mediating monocyte attachment and adhesion by exposing binding sites for specific counter- receptors on the monocyte surface. The central premise of this proposal is that engagement of specific attachment receptors on monocytes is an important pathway of monocyte activation, and that attachment-induced activation triggers a unique intracellular signalling pathway that has particular relevance to atherogenesis. Preliminary data has been obtained demonstrating that when peripheral blood monocytes are co-cultured with cytokine-activated endothelial cells a specific phenotypic change is induced in the monocytes that includes surface expression of procoagulant tissue factor, increased surface expression of CD36 (a glycoprotein scavenger receptor and adhesion receptor), and secretion of the pro- inflammatory cytokine TNF. This phenotypic change is the result of increased transcription of specific genes and requires direct contact between the endothelial cells and the monocytes. Engagement of E-selectin receptors on monocytes has been shown to play a major role in inducing these phenotypic changes. Plans are outlined to define the surface, cytoplasmic and nuclear signalling pathways involved in monocyte activation by cellular attachment receptors. In particular the role of E- selectin ligands (ES-1 and CD15) will be explored using immuno-inhibition and anti-sense oligonucleotide approaches. Attachment of monocytes to E- selectin transfected cells will be used to explore specific intracytoplasmic signalling pathways and intranuclear gene regulation mechanisms induced by E-selectin engagement. These studies will involve close interactions with Dr. D. Hajjar and B. Hempstead who are experts in cell signalling pathways. Specific genes activated by this pathway will be characterized by northern analysis, PCR, and ELISA with particular attention to monocyte/macrophage effector functions relevant to vascular biology. This will involve close interactions with Dr. D. Hajjar and A. Marcus looking at scavenger receptors, cytokines, and eicosanoids; and Dr. K. Hajjar looking at regulators of coagulation and fibrinolysis. Novel genes activated by E-selectin engagement will be identified by PCR differential display and/or positive selection cloning techniques. It is expected that the proposed studies may provide novel mechanistic insight into cell signalling and ultimately may allow development of novel therapeutic strategies for vascular and inflammatory diseases.
Keywords: biological signal transduction, blood coagulation, cell adhesion, cell adhesion molecule, molecular pathology, receptor binding, vascular endothelium, CD antigen, cytokine, gene expression, leukocyte activation /transformation, monocyte, receptor expression, selectin, enzyme linked immunosorbent assay, human subject, laboratory rabbit, mixed tissue /cell culture, molecular cloning, northern blotting, polymerase chain reaction, transfection
MOLECULAR AND CELL BIOLOGY OF CD36 A TSP RECEPTOR
Roy L Silverstein, Chairman
Medicineweill Medical College Of Cornell Univ
1300 York Avenue
new York, Ny 10021
Grant 5R01HL042540-09 from National Heart, Lung, And Blood Institute IRG: HEM
Abstract: CD36 is a multifunctional 88kD transmembrane glycoprotein expressed on platelets, monocytes, macrophages, certain specialized epithelial cells, and adipocytes. It has been shown to function as a cellular receptor for thrombospondin (TSP) and has recently been implicated as a macrophage scavenger receptor for oxidized low density lipoprotein (OxLDL). Oxidation of LDL is strongly linked to the pathogenesis of atherosclerosis in that lipid accumulation in atheromatous plaque results directly from internalization of OxLDL by intimal macrophages. In addition, OxLDL interactions with vascular cells may result in cellular activation to a prothrombotic, proatherogenic phenotype. The central hypothesis of this proposal is that CD36 is a major macrophage receptor for binding and internalizing ligands, including TSP and OxLDL. The focus will be to define the molecular basis of CD36 function and to probe its role in atherosclerosis. Studies in specific aim l will define structure-function relationships that govern CD36 receptor-ligand binding. Interactions with TSP and Oxidized LDL will be studies. These experiments will utilize recombinant CD36 peptides and solid phase and cellular binding studies. Specific aim 2 will define the mechanisms by which human macrophage CD36 internalizes ligands. Regulation of macrophage CD36 mRNA synthesis and surface protein expression in response to mediators of atherogenesis and differentiation will be studied using RNAse protection assays and immunomicroscopy. CD36-mediated internalization pathways and structure-function relationships that govern internalization will also be probed. Aim 3 will evaluate the role of CD36 in monocyte/macrophage biology by developing and studying a murine model genetically engineered so as not to express CD36. Completion of this project will increase understanding of early atherogenic events and allow for the development of novel therapeutic strategies by specific blockade of OxLDL-macrophage interactions. Furthermore, understanding how internalization and intracellular signaling is accomplished by CD36 may lead to broader insight into cellular function. The animal model may also be useful in testing the role of oxidants and anti-oxidants in atherogenesis
Keywords: CD antigen, low density lipoprotein, macrophage, receptor, receptor binding, thrombospondin atherosclerotic plaque, chimeric protein, genetic model, intermolecular interaction, ligand, low density lipoprotein receptor, membrane protein, model design /development, monocyte, protein structure /function, receptor expression RNase protection assay, immunocytochemistry, laboratory mouse, laboratory rabbit, transgenic animal
Project start date: 1989-07-01
Project end date: 2002-07-31
5R01HL042540-09 (2000): $287503
5R01HL042540-08 (1999): $281226
3R01HL042540-08S1 (1999): $35957
3R01HL042540-07S1 (1999): $2237
5R01HL042540-07 (1998): $242874
2R01HL042540-06A2 (1997): $237896
ROLE OF CD 36 AS PRO-THROMBOTIC PLATELET SURFACE RECEPTOR
Roy L Silverstein, Dept Chairman
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Abstract: Arterial thrombosis is a common complication of many systemic diseases, including atherosclerosis, diabetes, cancer, and chronic inflammatory conditions. This proposal will test the hypothesis that CD36 mediates platelet "hyper-reactivity" associated with systemic diseases, and thus contributes to arterial thrombosis. CD36 is a "pattern recognition" or scavenger receptor that recognizes ligands such as oxidized LDL, glycated proteins, and apoptotic cell membranes that are generated during pathological conditions. It is thus uniquely positioned to "sense" disease. Corollaries to the hypothesis are that platelets sensitized by CD36-ligand interactions may be resistant to anti-platelet therapies, and that levels of platelet CD36 expression, perhaps under the influence of genetic polymorphisms, may contribute to human thrombotic risk. Three specific aims are proposed. The first will utilize in vitro assays of human platelet function to characterize the role of CD36 in platelet activation. Whether CD36 ligands, including oxidized phospholipids, apoptotic cells, advanced glycation end products, and anti-CD36 autoantibodies can sensitize platelets to activation by low concentrations of other agonists will be determined, as will mechanisms by which CD36 initiates platelet signaling cascades. The second aim will characterize the pro-thrombotic role of CD36 in vivo using murine models. CD36 null mice crossed to an apoE null background will be used as a model of an atherogenic state, and CD36 nulls rendered hyperglycemic will be used as a model of diabetes. The effect of pharmacologic upregulation of CD36 will also be tested. The third aim will determine the potential role of human CD36 polymorphisms in athero-thrombosis. Variance in levels of human platelet surface CD36 expression among individuals will be correlated with CD36 genotype, and associations of CD36 genotype and platelet CD36 expression with thrombotic risk and platelet resistance to aspirin and/or clopridogel will be determined
Keywords: 2-(Acetyloxy)benzoic Acid; APOE [{C0003595}]; Acetylsalicylic Acid; Advanced Glycosylation End Products; Agonist; Apo-E; ApoE; Apolipoprotein E; Apoptotic; Aspergum; Aspirin; Atheroscleroses; Atherosclerosis; Atherosclerotic Cardiovascular Disease; Autoantibodies; Bizzozero`s corpuscle/cell; Blood Platelets; CD36; CD36 gene; Cancers; Cell Communication and Signaling; Cell Signaling; Cell membrane; Cells; Chronic; Complication; Cytoplasmic Membrane; Deetjeen`s body; Diabetes Mellitus; Disease; Disorder; Ecotrin; Empirin; Entericin; Extren; GP3B; GP4; GPIV; Genetic Polymorphism; Genotype; Glycation End Products, Advanced; Glycosylation End Products, Advanced; Hayem`s elementary corpuscle; Human; Human, General; Hyperglycemia; Individual; Inflammatory; Intracellular Communication and Signaling; Investigators; Knockout Mice; Ligands; Malignant Neoplasms; Malignant Tumor; Mammals, Mice; Man (Taxonomy); Man, Modern; Marrow platelet; Measurin; Mediating; Mice; Mice, Knock-out; Mice, Knockout; Modeling; Murine; Mus; Null Mouse; OxLDL; Pattern Recognition; Pattern Recognition/Display/Analysis; Phosphatides; Phospholipids; Plasma Membrane; Platelet Activation; Platelets; Polymorphism (Genetics); Polymorphism, Genetic; Position; Positioning Attribute; Productivity; Programs (PT); Programs [Publication Type]; Proteins; Receptor Protein; Research Personnel; Researchers; Resistance; Reticuloendothelial System, Platelets; Risk; Role; SCARB3; Signal Transduction; Signal Transduction Systems; Signaling; Surface; Systemic disease; Testing; Thrombocytes; Thrombosis; Up-Regulation; Up-Regulation (Physiology); Upregulation; acetylated LDL receptor; advanced glycation endproduct; atheromatosis; atherosclerotic vascular disease; autoimmune antibody; biological signal transduction; diabetes; disease/disorder; gene product; hyperglycemic; in vitro Assay; in vivo; malignancy; neoplasm/cancer; ox-LDL; oxidized LDL; oxidized low density lipoprotein; plasmalemma; polymorphism; programs; receptor; receptor, acetyl-LDL; resistant; scavenger receptor; self reactive antibody; social role; thrombocyte/platelet
Budget start date: 1-MAR-2010
Budget end date: 28-FEB-2011
5P50HL081011-05_0001 (2010): $396832
Sponsored Links Excellgen http://Excellgen.com
GENETIC AND CELLULAR DETERMINANTS OF ARTERIAL THROMBOSIS
Roy L Silverstein, Dept Chairman
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Grant 5P50HL081011-05 from National Heart, Lung, And Blood Institute
Abstract: Arterial thrombosis as the proximate cause of myocardial infarction, stroke, and limb gangrene remains the most important cause of mortality and morbidity in the USA. Blood platelets play a critical role in initiating arterial thrombi. The overall goal of this SCCOR is to discover genetic and cellular determinants of altered platelet function in systemic inflammatory states, such as atherosclerosis and diabetes. We have assembled a multidisciplinary team of clinical, basic and translational scientists and developed a Center with 6 scientific projects and 6 cores. Project 1 will use in vitro assays, in vivo mouse genetic models, and human genetic studies to examine the novel hypothesis that platelet glycoprotein CD36 mediates platelet hyper-reactivity associated with atherosclerosis, diabetes, and inflammation. Project 2 will explore a critically important clinical problem, platelet resistance to pharmacologic interventions, and will utilize a targeted human genetic approach coupled to sophisticated in vitro assessment of platelet function. Project 3 will explore the mechanistic role of platelet activating factor (PAF) in mediating the interface between inflammation and thrombosis. Project 4 will study a clinical population of patients undergoing endovascular carotid stenting as a model of the vulnerable plaque, focusing on developing novel imaging tools and biomarkers. Project 5 will study leukocyte-platelet interactions, focusing on the role of the Mac-1 integrin in regulating thrombosis. Project 6 will study patients with anti-phospholipid syndrome, an autoimmune disease that is an important cause of thrombosis, especially in young women, to characterize determinants of thrombotic risk. Core A will provide administrative support, Core B biostatistics support and Core C an infrastructure to support the clinical research components of all 6 projects. Core D will provide high throughput human genotyping services and Core E flow cytometry and mass spectrometry services. Core F will provide a curriculum and career development support for physicians training in clinical thrombosis research in SCCOR laboratories
Project start date: 2006-03-24
Project end date: 2011-02-28
Budget start date: 1-MAR-2010
Budget end date: 28-FEB-2011
PFA/PA: RFA-HL-04-016
5P50HL081011-05 (2010): $2700496
5P50HL081011-02 (2007): $2520364
1P50HL081011-01 (2006): $2530472
INSTITUTIONAL RESEARCH FELLOWSHIP AWARD IN HEMATOLOGY
Roy L Silverstein, Chairman
Weill Medical College Of Cornell Univ 1300 York Avenue New York, Ny 10021
Grant 5T32HL007029-29 from National Heart, Lung, And Blood Institute IRG: ZHL1
Project start date: 1975-07-01
Project end date: 2005-06-30
5T32HL007029-29 (2003): $109971
5T32HL007029-28 (2002): $309544
5T32HL007029-27 (2001): $118203
2T32HL007029-26 (2000): $268998
Sponsored Links Excellgen http://Excellgen.com
5T32HL007029-24 (1998): $188293
5T32HL007029-23 (1997): $188062
MOLECULAR MECHANISMS OF RPE PHAGOCYTOSIS--ROLE OF CD36
Roy L Silverstein, Chairman
Weill Medical College Of Cornell Univ 1300 York Avenue New York, Ny 10021
Grant 5R01EY010967-02 from National Eye Institute IRG: VISC
Project start date: 1995-05-01
Project end date: 1999-04-30
5R01EY010967-02 (1996): $232973
INSTITUTIONAL RESEARCH FELLOWSHIP AWARD IN HEMATOLOGY
Roy L Silverstein, Chairman
Weill Medical College Of Cornell Univ 1300 York Avenue New York, Ny 10021
Grant 5T32HL007029-22 from National Heart, Lung, And Blood Institute IRG: RTR
Project start date: 1975-07-01
Project end date: 2000-06-30
5T32HL007029-22 (1996): $187168