OXIDIZED PHOSPHOLIPIDS IN VASCULAR PATHOBIOLOGY
L Roy, Dept Chairman
Cleveland Clinic Lerner Col/med-cwrucity: Cleveland country: United States (us)
Grant 5P01HL087018-05 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-2011
Budget end date: 30-APR-2012
5P01HL087018-05 (2011): $2257974
Sponsored Links Excellgen http://Excellgen.com
L Roy, Dept Chairman
Cleveland Clinic Lerner Col/med-cwrucity: Cleveland country: United States (us)
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: ing; Animal Experimentation; Biometry; Blood Vessels; Budgets; Communication; Communities; Contract Services; Electronics; Equipment; Health Insurance Portability and Accountability Act; Human; human subject; Individual; Mails; Manuscripts; Medical; meetings; Mus; Oral; Phospholipids; posters; Preparation; Press Releases; Process; Productivity; Program Reviews; programs; Protocols documentation; Publishing; Quality Control; Radiation; Regulation; Research; Research Personnel; Safety; Schedule; Series; symposium; Telephone; Translating; Visit
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
5P01HL087018-05_9003 (2011): $135850
INFLAMMATORY CELL SIGNALING BY CD36
L Roy, Dept Chairman
Cleveland Clinic Lerner Col/med-cwrucity: Cleveland country: United States (us)
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: Address; Adipocytes; Apolipoprotein E; Apoptotic; Arterial Fatty Streak; atherogenesis; Atherosclerosis; Blood Vessels; CD36 gene; cell motility; Cells; Chronic; Complementary DNA; Complex; Data; Development; Diabetes Mellitus; Disease; Elements; Foam Cells; Goals; Grant; Human; Immune system; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Insulin Receptor; Insulin Resistance; interest; Knockout Mice; Lesion; Ligands; Link; Lipids; Lipoproteins; Low-Density Lipoproteins; macrophage; macrophage scavenger receptors; MAPK8 gene; Mediating; Mediator of activation protein; Metabolic Pathway; Metabolic syndrome; migration; Mitogen-Activated Protein Kinases; Molecular; monocyte; Mouse Strains; Mus; Obesity; oxidized lipid; particle; Pathogenesis; Pathway interactions; Phospholipids; Play; Process; programs; Publishing; Reagent; receptor; receptor function; Receptor Signaling; response; RNA Interference; Role; scavenger receptor; Signal Pathway; Signal Transduction; Technology; uptake
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
5P01HL087018-05_0004 (2011): $391986
Grants awarded to L Roy
CAM HERBAL COCKTAILS FOR BOOSTING ANTIOXIDENT ACTIVITY TO TREAT LIVER DISEASE
L Roy, Clinical Assistant Professor
University Of North Carolina Chapel Hillcity: Chapel Hill country: United States (us)
Grant 5R21AT003892-03 from National Center For Complementary & Alternative Medicine
Abstract: This application concerns the exploration of a novel strategy to increase the normally low bioavailability of polyphenolic antioxidants contained in botanical products by using knowledge of their metabolism to target specific herbal-herbal interactions. A major limitation of many prior clinical investigations is their use of inadequate oral dose regimens that failed to account for the rapid elimination and low bioavailability of polyphenolic antioxidants that results in low steady-state exposures to these potentially protective compounds and presumably accounts for their lack of efficacy. The rationale design of CAM herbal cocktails should result in dramatic increases in the steady-state concentrations of these potent antioxidants in additive or synergistic pharmacodynamic effects that may be common or unique for each of the botanical products contained in the cocktail. More than 50% of patients with chronic hepatitis C either have contraindications or do not achieve sustained virological response to therapy and would potentially benefit from the availability of alternative medicines. Our hypothesis is that the coadministration of silymarin with a CAM cocktail containing a structurally diverse mixture of polyphenolic natural products will boost silymarin´s antioxidant activity either through pharmacokinetic interactions that will slow elimination pathways for silymarin, or through pharmacodynamic interactions that result from the antioxidant or other beneficial properties of the CAM cocktail that are additive or synergistic with silymarin. The goal of the proposed double blind, placebo controlled, randomized clinical trial is to describe the pharmacokinetics, and to demonstrate the safety, tolerability, and antioxidant activity of a CAM herbal cocktail in patients with chronic hepatitis C genotype 1 infection and who have not received prior peginterferon-based therapies. In Specific Aim 1 we will perform a pharmacokinetic study that will determine the effects of a CAM herbal cocktail containing milk thistle and green tea extracts on the exposures of patients to major silymarin flavonolignans. For Specific Aim 2, we will identify and use an optimal procedure for the assay of 8-isoprostane F2, as a measure of oxidative stress, to determine the antioxidant activity of the CAM herbal cocktail in patients with chronic HCV. The goal of this study is to provide an alternative medicine for patients with hepatitis C where treatment options are unavailable. Regardless of the positive public perceptions regarding the safety and efficacy of botanical products, data on the use of high doses of herbal combinations to optimize health outcomes are lacking and vigilance regarding adverse effects is warranted. The goal of this proposal is to evaluate the safety, tolerability, and effectiveness of potentially beneficial herbal-herbal interactions using high dose herbal cocktails for the treatment of liver diseases where treatment options are not available
Keywords: Accounting; active control; Address; Adverse effects; Affinity; Alternative Medicine; Antioxidants; Area Under Curve; Attenuated; base; Biological; Biological Assay; Biological Availability; Biological Factors; Black Tea; Blood; Botanicals; Camellia sinensis (Plant); Catechin; Characteristics; Chronic; Chronic Hepatitis C; Clinical; Clinical Trials; cohort; Complex; Compliance behavior; Control Groups; Coronary Arteriosclerosis; cost; Data; design; Diabetes Mellitus; Disease; Dose; Double-Blind Method; Drug Kinetics; effective therapy; Effectiveness; efficacy trial; Enzyme-Linked Immunosorbent Assay; Epigallocatechin Gallate; Etiology; Exhibits; Exposure to; Failure (biologic function); Fibrosis; Flavonoids; Flavonolignans; Frequencies (time pattern); Future; Genotype; Glucuronides; Goals; Green tea (dietary); Half-Life; Health; Hepatic; Hepatitis C; Hepatitis C virus; human disease; Infection; Inorganic Sulfates; isoprostaglandin F2alpha type-III; Knowledge; Literature; Liver; Liver diseases; liver metabolism; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Measures; Metabolism; Milk Thistle; Multicenter Trials; novel strategies; Oral; Oral Administration; Outcome; Oxidative Stress; Pathway interactions; Patients; Perception; Pharmacodynamics; Phase; pill (pharmacologic); Placebo Control; Plasma; polyphenol; Procedures; Production; Property; public health relevance; Publishing; Randomized Clinical Trials; Reactive Oxygen Species; Regimen; Relative (related person); Research Design; response; Risk; Route; Safety; Sampling; Schedule; Serum; silibinin; Silymarin; Site; Source; Standard Preparations; Time; UGT1A1 enzyme; UGT1A1 gene; Unspecified or Sulfate Ion Sulfates; uptake; Urine; Variant; vigilance; Virus Diseases
Project start date: 2008-09-30
Project end date: 2012-06-30
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2012
PFA/PA: PA-06-510
5R21AT003892-03 (2010): $237949
REGULATION OF THE ANTI-ANTIANGIOGENIC SWITCH BY CD36, THROMBOSPONDIN, AND HRGP
L Roy, Dept Chairman
Cleveland Clinic Lerner Col/med-cwrucity: Cleveland country: United States (us)
Grant 5R01HL085718-04 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: Address; angiogenesis; Angiogenesis Inhibitors; Angiogenic Factor; Angiogenic Proteins; Angiogenic Switch; base; Binding (Molecular Function); Biological; Blood; Blood capillaries; Blood Vessels; CD36 gene; Cell Line; Cells; design; Development; Diabetes Mellitus; Diabetic Retinopathy; Disease; Down-Regulation; Endothelial Cells; Genes; Goals; Growth; Health; histidine-rich glycoprotein; Human; human disease; In Vitro; in vivo; Inflammation; Laboratories; Lead; Ligands; lysophosphatidic acid; Lysophospholipids; Malignant Neoplasms; Measures; Mediating; Messenger RNA; Mus; Myocardial Infarction; NMR Spectroscopy; novel therapeutic intervention; novel therapeutics; Organism; overexpression; Phospholipids; Phosphorylation; Post-Translational Protein Processing; Process; prognostic; Protein Kinase C; Proteins; Radiation; receptor; recombinant peptide; Regulation; Regulatory Pathway; response; Role; Signal Transduction; Skin; stroke; Surface; System; Thrombospondin 1; Thrombospondins; Tissues; Transgenic Mice; Transgenic Organisms; tumor growth; Ultraviolet B Radiation; Ursidae Family
Project start date: 2008-06-20
Project end date: 2013-05-31
Budget start date: 1-JUN-2011
Budget end date: 31-MAY-2012
PFA/PA: PA-07-070
5R01HL085718-04 (2011): $392500
PPAR GAMMA AND NOX4 IN PULMONARY HYPERTENSION
L Roy
Emory Universitycity: Atlanta country: United States (us)
Grant 1R01HL102167-01A1 from National Heart, Lung, And Blood Institute
Abstract: Despite existing therapies, pulmonary hypertension (PH) causes significant morbidity and mortality. This proposal focuses on peroxisome proliferator-activated receptor gamma (PPARg) as a new target in PH therapy. Evolving evidence demonstrates that chronic hypoxia and other causes of PH are associated with increased expression and activity of the NADPH oxidase, Nox4. Nox4 generates reactive oxygen species that contribute to vasoconstriction, pulmonary vascular cell proliferation, and PH pathogenesis. Stimulating PPARg with thiazolidinedione ligands reduces the expression and activity of Nox4 and attenuates hypoxia-induced vascular remodeling, right ventricular hypertrophy, and pulmonary hypertension in a mouse model. Preliminary data confirm that Nox4 is upregulated in endothelial cells from patients with idiopathic pulmonary arterial hypertension. Therefore, this proposal examines the hypothesis that activation of PPARg provides a novel strategy to attenuate hypoxia-induced Nox4 expression, oxidative stress, vascular remodeling and PH. To explore this hypothesis, Aim 1 will examine the role of Nox4 in hypoxia-induced PH and its regulation by PPARg using endothelial- and smooth muscle-targeted Nox4 knockout mice. Aim 2 will use endothelial- and smooth muscle-targeted PPARg knockout or overexpressing mice to define pulmonary vascular cell compartments that are critical for PPARg ligand-induced alterations in Nox4 and PH. Aim 3 will examine the molecular mechanisms by which PPARg activation attenuates Nox4 expression in the pulmonary vasculature. In vitro studies will be performed using hypoxia-exposed human pulmonary artery smooth muscle or endothelial cells. The long-term goals of this proposal are to define mechanisms by which PPARg activation attenuates PH and to facilitate the development of new PH therapy. High blood pressure in the lung, also called pulmonary hypertension, is a devastating condition that affects many patients and for which there are no effective therapies. This proposal explores a new type of therapy for pulmonary hypertension using an animal model by examining not only the efficacy of this new treatment strategy but also the basic mechanisms by which it works. These studies have the potential to identify novel treatment strategies that could be applied to patients with pulmonary hypertension
Keywords: 2, 4-thiazolidinedione; activating transcription factor; Active Oxygen; Affect; Animal Model; Animal Models and Related Studies; Animals; ATF; Attenuated; Binding; Binding (Molecular Function); biological signal transduction; Blood Pressure, High; Blood Vessels; Bone-Derived Transforming Growth Factor; Cell Communication and Signaling; Cell Growth in Number; Cell Multiplication; Cell Proliferation; Cell Signaling; Cell Wall; Cells; Cellular Proliferation; Chronic; Data; Development; Disease; disease/disorder; Disorder; effective therapy; Endothelial Cells; Experimental Models; Experimental Models, Other; Glitazones; Goals; HIF 1; HIF-1 protein; HIF1; HIF1 protein; Human; Human, General; hyperpiesia; hyperpiesis; Hypertension; hypertensive disease; Hypoxia; hypoxia inducible factor 1; Hypoxic; In Vitro; in vivo; in vivo Model; Intracellular Communication and Signaling; Knock-out; Knockout; knockout animal; Knockout Mice; Leiomyocyte; Ligands; loss of function; Lung; Mammals, Mice; Man (Taxonomy); Man, Modern; Mediating; member; Mice; Mice, Knock-out; Mice, Knockout; Milk Growth Factor; model organism; Modeling; Models, Experimental; Molecular; Molecular Interaction; Morbidity; Morbidity - disease rate; Mortality; Mortality Vital Statistics; mouse model; Murine; Mus; Muscle, Involuntary; Muscle, Smooth; Myocytes, Smooth Muscle; NADPH Oxidase; new approaches; novel; novel approaches; novel strategies; novel strategy; Nuclear; Nuclear Hormone Receptor Superfamily; Nuclear Hormone Receptors; Null Mouse; overexpression; Oxidative Stress; Oxygen Deficiency; Oxygen Radicals; Pathogenesis; pathway; Pathway interactions; Patients; PDGF; Peroxisome Proliferative Activated Receptor Gamma; Peroxisome Proliferator-Activated Receptor gamma; Platelet Transforming Growth Factor; Platelet-Derived Growth Factor; PPAR gamma; PPARG; PPARG1; PPARG2; PPARgamma; Pro-Oxidants; Production; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); protective effect; public health relevance; Publishing; pulmonary; pulmonary arterial hypertension; Pulmonary Artery; Pulmonary artery structure; Pulmonary Hypertension; Reactive Oxygen Species; Regulation; Respiratory System, Lung; response; Right Ventricular Hypertrophy; Role; Signal Transduction; Signal Transduction Systems; Signaling; Smooth muscle (tissue); Smooth Muscle Cells; Smooth Muscle Myocytes; Smooth Muscle Tissue Cell; social role; TGF B; TGF-beta; TGFbeta; Therapeutic Effect; therapeutic target; thiazolidinedione; Thiazolidinedione Receptor; Thiazolidinediones; Transforming Growth Factor beta; treatment strategy; vascular; Vascular constriction (function); Vascular Hypertensive Disease; Vascular Hypertensive Disorder; Vascular remodeling; Vasoconstriction; Wild Type Mouse; Work
Relevance: High blood pressure in the lung, also called pulmonary hypertension, is a devastating condition that affects many patients and for which there are no effective therapies. This proposal explores a new type of therapy for pulmonary hypertension using an animal model by examining not only the efficacy of this new treatment strategy but also the basic mechanisms by which it works. These studies have the potential to identify novel treatment strategies that could be applied to patients with pulmonary hypertension
Project start date: 2011-01-01
Project end date: 2014-12-31
Budget start date: 1-JAN-2011
Budget end date: 31-DEC-2011
PFA/PA: PA-10-067
1R01HL102167-01A1 (2011): $312154
5R01HL102167-02 (2012): $291982