SIGNALING MOLECULES IN ACUTE LUNG INJURY
Guy A Zimmerman, Professor Of Internal Medicine
University Of Utah 75 South 2000 East Salt Lake City, Ut 84112
Grant 5P50HL050153-050002 from National Heart, Lung, And Blood Institute
Abstract: Inflammatory mechanisms are involved in the initiation, amplification, propagation, and resolution of Adult Respiratory Distress Syndrome (ARDS). Neutrophils accumulate in the lungs of patients with ARDS, release granular enzymes and other toxins that mediate damage to cells of the alveolar-capillary membrane in vitro, and injure lungs of experimental animals in vivo. Similarly signaling molecules, such as interleukin and biologically-active lipids, that directly activate PMNs and induced mobilization of granular contents, generation of oxygen radicals, and other functional alterations are expressed by endothelium and other cells in ARDS. The identities of many of these signaling molecules remain unknown, and the mechanisms that cause such molecules to become expressed in a dysregulated, injurious fashion, as opposed to a homeostatic, protective fashion, remain undefined. Broad goals of this project are to characterize neutrophil signaling molecules of different classes, define the mechanisms that regulate their expression, and determine mechanisms of dysregulation. We will focus here on characterization of a signaling molecule that induces neutrophil degranulation, because this event appears to be of particular importance in the pathogenesis of ARDS. In preliminary experiments, we found that endothelial cells treated with the cytokines IL-1 and TNFa or with LPS, synthesize a signaling molecule that induces PMN to secrete lactoferrin, a specific granule marker, and elastase, primary granule protease, into the fluid phase. Maximal generation of the degranulating signal is dissociated from surface expression of molecules that mediate adhesion and transmigration of PMN. These features suggest that the degranulating factor is not involved in physiologic transmigration of PMN. These features suggest that the degranulating factor is not involved in physiologic targeting events. Furthermore, endothelial cell monolayers are disrupted when incubated with PMN in the presence of the degranulating activity, suggesting that release of elastase or other PMN proteases induced by this factor mediates endothelial injury. Our hypotheses are that human endothelial cells generate and release a novel signaling molecule that stimulates PMN degranulation. PMN granular proteases amplify injury to the alveolar capillary membrane and may be key mediators of vascular destruction that occurs in ARDS. Dysregulated expression of signaling molecules by endothelial cells is a fundamental pathophysiologic mechanism in acute lung injury. Our specific aims are 1) To purify, characterize, sequence, and clone the new degranulating factor. 2) To characterize the biologic activities of the degranulating factor. 3) To characterize the synthesis and secretion of the factor and define its relationship to other signaling molecules. 4) To determine if the degranulating factor is found in the blood or lung tissue of patients during the acute, subacute, and reparative phases of ARDS.
Keywords: adult respiratory distress syndrome, chemical structure /function, granule, leukocyte activation /transformation, neutrophil, vascular endothelium, biological signal transduction, cytokine, gene expression, inflammation, protein sequence, protein structure /function, secretion, selectin, human subject, molecular cloning, tissue /cell culture
Project start date: 1997-12-01
Project end date: 1998-11-30
Sponsored Links Excellgen http://Excellgen.com
Guy A Zimmerman, Professor Of Internal Medicine
University Of Utah 75 South 2000 East Salt Lake City, Ut 84112
Grant 5P50HL050153-109002 from National Heart, Lung, And Blood Institute
Abstract: The Cell Biology Core will interact with and support each of the projects in the SCOR. The central aim will be to provide well-characterize primary human cells and cell lines for use as models and in cell biologic studies that are relevant to lung injury in acute respiratory distress syndrome (ARDS), as we have done in the current funding period. Also, as in the current funding period. the major cell types will include freshly isolated neutrophils, monocytes, and other leukocytes and primary cultures of human endothelial cells. In addition, we will also grow, characterize, and furnish to the projects a variety of cell lines for specialized studies, including a pulmonary endothelial cell line developed in the Core. We focus on human cell types, because of immediate relevance, but also will provide cultured cells or established cell lines from experimental animals when useful. Core personnel also advise SCOR investigators on cell biology protocols, help identify and develop new preparations, pilot and develop specialized techniques, assays, and models, and in some cases perform specialized cell biologic studies or assays with or for SCOR investigators. The core will foster and extend fundamental investigations in the SCOR that will lead to new knowledge of the biologic features of lung injury in ARDS.
Keywords: adult respiratory distress syndrome, biomedical facility, cell line, tissue /cell culture, tissue resource /registry, human tissue
SIGNALING MOLECULES IN ACUTE LUNG INJURY
Guy A Zimmerman, Professor Of Internal Medicine
University Of Utah 75 South 2000 East Salt Lake City, Ut 84112
Grant 5P50HL050153-040002 from National Heart, Lung, And Blood Institute
Abstract: Inflammatory mechanisms are involved in the initiation, amplification, propagation, and resolution of Adult Respiratory Distress Syndrome (ARDS). Neutrophils accumulate in the lungs of patients with ARDS, release granular enzymes and other toxins that mediate damage to cells of the alveolar-capillary membrane in vitro, and injure lungs of experimental animals in vivo. Similarly signaling molecules, such as interleukin and biologically-active lipids, that directly activate PMNs and induced mobilization of granular contents, generation of oxygen radicals, and other functional alterations are expressed by endothelium and other cells in ARDS. The identities of many of these signaling molecules remain unknown, and the mechanisms that cause such molecules to become expressed in a dysregulated, injurious fashion, as opposed to a homeostatic, protective fashion, remain undefined. Broad goals of this project are to characterize neutrophil signaling molecules of different classes, define the mechanisms that regulate their expression, and determine mechanisms of dysregulation. We will focus here on characterization of a signaling molecule that induces neutrophil degranulation, because this event appears to be of particular importance in the pathogenesis of ARDS. In preliminary experiments, we found that endothelial cells treated with the cytokines IL-1 and TNFa or with LPS, synthesize a signaling molecule that induces PMN to secrete lactoferrin, a specific granule marker, and elastase, primary granule protease, into the fluid phase. Maximal generation of the degranulating signal is dissociated from surface expression of molecules that mediate adhesion and transmigration of PMN. These features suggest that the degranulating factor is not involved in physiologic transmigration of PMN. These features suggest that the degranulating factor is not involved in physiologic targeting events. Furthermore, endothelial cell monolayers are disrupted when incubated with PMN in the presence of the degranulating activity, suggesting that release of elastase or other PMN proteases induced by this factor mediates endothelial injury. Our hypotheses are that human endothelial cells generate and release a novel signaling molecule that stimulates PMN degranulation. PMN granular proteases amplify injury to the alveolar capillary membrane and may be key mediators of vascular destruction that occurs in ARDS. Dysregulated expression of signaling molecules by endothelial cells is a fundamental pathophysiologic mechanism in acute lung injury. Our specific aims are 1) To purify, characterize, sequence, and clone the new degranulating factor. 2) To characterize the biologic activities of the degranulating factor. 3) To characterize the synthesis and secretion of the factor and define its relationship to other signaling molecules. 4) To determine if the degranulating factor is found in the blood or lung tissue of patients during the acute, subacute, and reparative phases of ARDS.
Keywords: adult respiratory distress syndrome, chemical structure /function, granule, leukocyte activation /transformation, neutrophil, vascular endothelium, biological signal transduction, cytokine, gene expression, inflammation, protein sequence, protein structure /function, secretion, selectin, human subject, molecular cloning, tissue /cell culture
Project start date: 1996-12-01
Project end date: 1997-11-30
DYSREGULATED EXPRESSION OF SIGNALING MOLECULES IN ACUTE LUNG INJURY
Guy A Zimmerman, Professor Of Internal Medicine
University Of Utah 75 South 2000 East Salt Lake City, Ut 84112
Grant 5P50HL050153-100002 from National Heart, Lung, And Blood Institute
Abstract: A fundamental hypothesis in our Special Center of Research is that dysregulated cell-cell interactions initiate and/or amplify inflammatory injury to the alveolar capillary membrane in Acute Respiratory Distress. Syndrome (ARDS), a common and lethal cause of lung damage, and that endothelial cells are critically involved. Identification of signaling molecules and other factors dysregulated cell-cell interactions in ARDS is a central and long-term goal of this project. In the current application, we focus on signaling molecules for neutrophils synthesized by stimulated and injured human endothelial cells. Neutrophils (PMNs) initiate, amplify, and influence the maintenance and outcome of acute lung injury. Endothelial cells are the first structural cells that PMNs encounter in the inflamed and injured lung and, by virtue of their ability to synthesize signaling molecules on a variety of classes, are particular points of dysregulated information transfer to the leukocytes. The mechanisms that regulate the expression and biologic actions of endothelial signaling molecules of the same or different classes, the identities of some of these signaling factors, and the mechanisms by which they become dysregulated in inflammatory injury are largely unknown. This project addresses each of these issues in interrelated studies based on the above hypotheses and on observations and preliminary data generated in the current funding period. The first specific aim is to characterize the regulation of expression and actions of chemokines produced by human endothelial cells that signal neutrophil activation, focusing on ENA-78 and related family members. The second specific aim is to characterize new endothelial signaling factors that we have identified by screening cDNA libraries from stimulated endothelial cells and by subtractive hybridization. The third specific aim is to characterize expression and distribution of endothelial signaling molecules in lung tissue from patients with ARDS and in tissue from control and comparative subjects. The fundamental and correlative nature of these studies will yield new knowledge that may lead to novel strategies of prevention and therapy of ARDS.
Keywords: adult respiratory distress syndrome, cell cell interaction, chemokine, leukocyte activation /transformation, neutrophil, vascular endothelium, biological signal transduction, cytokine, gene expression, inflammation, clinical research, genetic library, human subject, laboratory mouse, molecular cloning, subtraction hybridization, tissue /cell culture
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Guy A Zimmerman
NEW PATHWAYS IN THROMBOSIS AND INFLAMMATION MEDIATED BY SEMAPHORIN-PLEXIN SIGNALI
Guy A Zimmerman
University Of Utah, 75 South 2000 East, Salt Lake City, Ut 84112
Grant 5RC1HL100121-02 from National Heart, Lung, And Blood Institute
Abstract: This application addresses broad Challenge Area (04), Clinical Research, and specific Challenge Topic 04-HL-103 assess the role of leukocyte interaction with platelets, erythrocytes, and endothelium in the pathogenesis of heart, lung, and blood diseases. Our proposal explores a new paradigm in molecular signaling and cell-cell interactions that is broadly relevant to human thrombotic and inflammatory diseases, and has the potential to generate novel therapeutic strategies and agents in addition to providing new knowledge in the field. The project unites a team of established investigators that is uniquely well-suited to examine clinically-applicable and basic questions relevant to the pathobiology of hemostasis and inflammation, builds on new discoveries that we have recently made, and poses focused aims that can be addressed expeditiously with expertise and technical capabilities that are in place in our collaborative group. The project is thus poised for rapid translational investigation and, potentially, fast track application. These features make it ideal for the Challenge Grant initiative. Thrombosis and inflammation are intricately linked in the pathogenesis of acute coronary syndromes, stroke, sepsis, acute lung injury, and a variety of other devastating human disorders. While much is known, there remain major gaps in our knowledge regarding the molecular pathways and cellular events that integrate inflammatory and hemostatic signaling in health, and mediate dysregulated signaling in disease. We have discovered a previously-unrecognized pathway that has these critical characteristics. Semaphorin-plexin D1 signaling has recently-identified roles in cellular guidance in the nervous system, but was not known to influence platelet activation, platelet-leukocyte interactions, or hemostatic and inflammatory events in vivo. Our preliminary data yield strong evidence that the semaphorin-plexin D1 signaling axis directly influences each of these, and that it may therefore be a novel target for molecular intervention. Because our current evidence indicates that semaphorin-plexin D1 signaling alters prothrombotic responses of activated platelets by modifying intracellular checkpoints and cytoskeletal organization distal to receptor-mediated cellular activation, new therapeutic agents that target this pathway could potentially be used independently or in combination with other antithrombotic therapies including cyclooxygenase inhibitors (aspirin), thienopyridines (clopidogrel), and anticoagulants (heparin, warfarin). The inter-related specific aims that we propose employ in vitro studies in informative human cell models, in vivo models carefully chosen for preclinical relevance and correlation with the in vitro experiments, and analyses of patient samples, and will rapidly advance our basic, translational, and clinical understanding of semaphorin-plexin signaling in hemostasis, inflammation, and vascular disease. The project will also form the basis for future investigations in many other experimental models and a variety of human syndromes. Our studies will exploit scientific and clinical opportunities, have the potential for broad and major impact, and will influence paradigms in diverse scientific and translational communities. Our proposal is thus directly responsive to the Challenge Grant goals and missions. The investigations outlined in this proposal will address cellular and biochemical mechanisms that contribute to thrombotic and inflammatory diseases, which are major public health problems. Unregulated thrombosis (clot formation, often occluding blood vessels) and inflammation contribute directly to heart attack, stroke, sepsis ("blood poisoning"), acute lung injury, and a host of other devastating human disorders. Many gaps in our understanding of these disease and disorders remain, limiting our ability to develop new and improved therapies and preventative measures. The molecular pathway that we have discovered and will examine in this proposal is a potential target for new and novel molecular therapies. Our studies will also provide invaluable new information on how clot formation and inflammation are controlled in health, and become uncontrolled and injurious in disease
Keywords: 1-(4`-Hydroxy-3`-coumarinyl)-1-phenyl-3-buta; 2-(Acetyloxy)benzoic Acid; 3-(Alpha-acetonylbenzyl)-4-hydroxycoumarin; 3-Alpha-phenyl-beta-acetylethyl-4-hydroxycoumarin; 4-Hydroxy-3-(3-oxo-1-phenylbutyl)-2H-1-benzopyran-2-one; Acetylsalicylic Acid; Acute Pulmonary Injury; Address; Adherence; Adherence (attribute); Adhesion Molecule; Age; Anticoagulant Agents; Anticoagulant Drugs; Anticoagulants; Apoplexy; Area; Aspergum; Aspirin; B2 antigen, Xenopus; Binding; Binding (Molecular Function); Biochemical; Biological; Biological Models; Bizzozero`s corpuscle/cell; Blood Diseases; Blood Platelets; Blood Poisoning; Blood Vessels; Blood erythrocyte; Blood leukocyte; Blood normocyte; COX inhibitor; Cardiac infarction; Cardiovascular; Cardiovascular Body System; Cardiovascular system; Cardiovascular system (all sites); Cell Adhesion Molecules; Cell Communication; Cell Communication and Signaling; Cell Interaction; Cell Signaling; Cell model; Cell-to-Cell Interaction; Cells; Cellular model; Cerebral Stroke; Cerebrovascular Apoplexy; Cerebrovascular Stroke; Cerebrovascular accident; Characteristics; Clinical; Clinical Research; Clinical Study; Clotting; Coagulation; Coagulation Process; Communication; Communities; Complex; Cues; Cyclooxygenase Inhibitors; Cytoskeletal Modeling; Cytoskeletal Organization; Cytoskeletal Organization Process; Cytoskeletal Reorganization; Data; Deetjeen`s body; Diagnosis; Disease; Disorder; Distal; Ecotrin; Empirin; Endothelium; Entericin; Erythrocytes; Erythrocytic; Event; Experimental Models; Experimental Models, Other; Extracellular Matrix, Integrins; Extren; Family; Family member; Future; Gene Expression; Goals; Grant; Hayem`s elementary corpuscle; Health; Heart; Hematologic Diseases; Hematological Disease; Hematological Disorder; Hemostasis; Hemostatic Agents; Hemostatic function; Hemostatics; Heparin; Heparinic Acid; Human; Human, General; INFLM; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Inhibitors, Cyclo-Oxygenase; Integrins; Intervention; Intervention Strategies; Intracellular Communication and Signaling; Investigation; Investigators; Knowledge; Lead; Leukocytes; Ligands; Link; Lung; Lung Injury, Acute; Mammals, Mice; Man (Taxonomy); Man, Modern; Marrow erythrocyte; Marrow leukocyte; Marrow platelet; Measures; Measurin; Mediating; Mediator; Mediator of Activation; Mediator of activation protein; Messenger RNA; Mice; Mission; Model System; Models, Biologic; Models, Experimental; Molecular; Molecular Interaction; Molecular Target; Murine; Mus; Myocardial Infarct; Myocardial Infarction; NRVS-SYS; Nerve Cells; Nerve Unit; Nervous System; Nervous system structure; Neural Cell; Neurocyte; Neurologic Body System; Neurologic Organ System; Neurons; Organ System, Cardiovascular; Pathogenesis; Pathologic; Pathway interactions; Patients; Pattern; Pb element; Pharmaceutical Agent; Pharmaceuticals; Pharmacologic Substance; Pharmacological Substance; Platelet Activation; Platelets; Prostaglandin Endoperoxide Synthase Inhibitors; Prostaglandin Synthase Inhibitors; Prostaglandin Synthesis Antagonists; Public Health; RNA, Messenger; Receptor Protein; Recombinants; Red Blood Cells; Red Cell; Red blood corpuscule; Red cell of marrow; Research Personnel; Researchers; Respiratory System, Lung; Reticuloendothelial System, Erythrocytes; Reticuloendothelial System, Leukocytes; Reticuloendothelial System, Platelets; Role; Sampling; Semaphorins; Sepsis; Septicemia; Severities; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Signaling Molecule; Stroke; Syndrome; Technical Expertise; Testing; Therapeutic Agents; Thrombocytes; Thrombosis; Thrombus; Vascular Accident, Brain; Vascular Diseases; Vascular Disorder; Vascular, Heart; Warfarin; White Blood Cells; White Cell; acute coronary syndrome; acute lung injury; axon growth; axonal growth; base; biological signal transduction; blood corpuscles; blood disorder; blood thinner; blood vessel disorder; bloodstream infection; brain attack; cardiac infarct; cell adhesion protein; cerebral vascular accident; circulatory system; clopidogrel; coronary attack; coronary infarct; coronary infarction; disease/disorder; experiment; experimental research; experimental study; healthy volunteer; heart attack; heart infarct; heart infarction; heavy metal Pb; heavy metal lead; improved; in vivo; in vivo Model; interventional strategy; mRNA; meetings; neuronal; neuronal guidance; new therapeutics; next generation therapeutics; novel; novel therapeutics; pathway; plexin; pre-clinical; preclinical; preclinical study; protein expression; public health medicine (field); pulmonary; receptor; research study; response; septic; septicaemia; social role; stroke; thienopyridine; thrombocyte/platelet; thrombopoiesis inhibitor; vascular; white blood cell; white blood corpuscle
Relevance: The investigations outlined in this proposal will address cellular and biochemical mechanisms that contribute to thrombotic and inflammatory diseases, which are major public health problems. Unregulated thrombosis (clot formation, often occluding blood vessels) and inflammation contribute directly to heart attack, stroke, sepsis ("blood poisoning"), acute lung injury, and a host of other devastating human disorders. Many gaps in our understanding of these disease and disorders remain, limiting our ability to develop new and improved therapies and preventative measures. The molecular pathway that we have discovered and will examine in this proposal is a potential target for new and novel molecular therapies. Our studies will also provide invaluable new information on how clot formation and inflammation are controlled in health, and become uncontrolled and injurious in disease
Project start date: 2009-09-30
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: RFA-OD-09-003
5RC1HL100121-02 (2010): $498264
1RC1HL100121-01 (2009): $498737
CELLULAR AND MOLECULAR RESPONSES IN LUNG DISEASE
Guy A Zimmerman, Professor Of Internal Medicine
University Of Utah 75 South 2000 East Salt Lake City, Ut 84112
Grant 5T32HL007636-15 from National Heart, Lung, And Blood Institute IRG: ZHL1
Abstract: The primary goal of this proposed program is to train young physician- scientists for a full-time academic career in pulmonary research. The emphasis will be placed on developing and cultivating highly qualified trainees, allowing them to acquire the necessary knowledge and skills required to evolve as independent investigators. The research will be conducted under the personal preceptorship of active senior investigators in the departments of Medicine, Pediatrics, Biology, Biochemistry, Pathology or Human Genetics at the University of Utah. The training program utilizes established strengths at the University of genetics and molecular biology while the patient-oriented research is aligned with ongoing investigations in acute lung injury, in particular the Acute Respiratory Distress Syndrome (ARDS). Our research training philosophy is that such training can best be achieved on the basis of the most direct day-to-day interaction between trainees and faculty preceptors who have demonstrated success in preparing individuals for careers in research. A highly conducive environment for a multi- disciplinary approach exists due to the close collaborative ties already developed among various preceptors and evolving associations and collaborations. The preceptorial research training will be supplemented with formal course work, lectures, conferences, seminars, and other pertinent educational modalities in order to prepare the trainees to formulate innovative and meritorious research programs in pulmonary diseases at the conclusion of their training. The proposed preceptors represent a group of active scientists engaged in research in five interrelated thematic areas relevant to lung health and disease. Each preceptor has an NIH-funded productive research program. The thematic areas include 1) Leukocyte Biology; 2) Vascular Cell Biology; 3) Developmental Biology and Aging; 4) Human Genetic Diseases; and 5) Patient-Oriented Research. We believe the program will enable the trainees to master the complexities of modern biomedical research, and will provide an appreciation for the collaborative interdisciplinary approach required for research into medical problems.
Project start date: 1994-07-15
Project end date: 2005-06-30
5T32HL007636-15 (2003): $10144
MOLECULAR INTERACTIONS--MYELOID CELLS W ENDOTHELIUM
Guy A Zimmerman, Professor Of Internal Medicine
University Of Utah 75 South 2000 East Salt Lake City, Ut 84112
Grant 5R01HL044525-14 from National Heart, Lung, And Blood Institute IRG: PTHA
Abstract: The long term goal of this project is to understand how adhesive interactions induce and integrate intercellular signaling between monocytes and neutrophils, which are leukocytes of the myeloid lineage, and endothelial cells. We also will determine how these interactions influence subsequent functional responses of the leukocytes in homeostatic inflammation and in inflammatory disease. The change in title of the project from "Molecular Mechanisms of Neutrophil Adhesion to Endothelium" to "Molecular Interactions of Myeloid Cells with Endothelium" reflects a larger scope, including a focus on adhesion-dependent signaling mechanisms and new investigations of the interactions of monocytes with endothelium, in addition to studies of PMNs. Specific adhesion molecules tether endothelial cells and myeloid cells together; similarly, such tethering molecules can bind platelets to myeloid cells and can mediate adhesion between the myeloid leukocytes themselves. We will determine how these adhesive interactions lead to the generation of intracellular signals that alter critical inflammatory functions of the myeloid cells. We will focus on expression of immediate-early genes, on cell spreading, and on related cellular responses. We will determine how molecules known to tether myeloid leukocytes transmit signals, and orchestrate them when acting in concert with other factors. Using isolated myeloid leukocytes and cultured endothelium, transfected cells, model membranes and purified molecules, we will characterize the roles of selectins, in Specific Aims l and 2. In Specific Aim 3, we will use similar strategies to identify tethering and signaling mechanisms of ICAM-3, a new member of the ICAM family, in myeloid cell interactions. In Specific Aim 4, we will examine "outside-in" signaling by beta32 (CD11/CD18) integrins in myeloid cells using two traditional approaches, adhesion of leukocytes to specific ligands and incubation with antibodies that may induce functional alterations, and a novel approach - study of myeloid cells with targeted deletions of individual CD11/CD18 heterodimers. Information from these reduced systems will then be used to analyze complex models of inflammation (cell-cell interactions in vitro) and, ultimately, to identify new strategies for understanding and modifying inflammatory events in vivo and in humans.
Keywords: atherosclerosis, biological signal transduction, inflammation, leukocyte adhesion molecule, monocyte, neutrophil, CD antigen, cell cell interaction, dimer, gene expression, glycoprotein, homeostasis, integrin, membrane protein, molecular biology, protein purification, selectin, vascular endothelium, human tissue, laboratory rabbit, polymerase chain reaction, tissue /cell culture, transfection
Project start date: 1990-04-01
Project end date: 2004-03-31
5R01HL044525-14 (2003): $318715
5R01HL044525-13 (2002): $309538
5R01HL044525-12 (2001): $300721
5R01HL044525-11 (2000): $292250
MOLECULAR INTERACTIONS OF MYELOID CELLS WITH ENDOTHELIUM
Guy A Zimmerman, Professor Of Internal Medicine
University Of Utah 75 South 2000 East Salt Lake City, Ut 84112
Grant 5R01HL044525-09 from National Heart, Lung, And Blood Institute IRG: PTHA
Abstract: The long term goal of this project is to understand how adhesive interactions induce and integrate intercellular signaling between monocytes and neutrophils, which are leukocytes of the myeloid lineage, and endothelial cells. We also will determine how these interactions influence subsequent functional responses of the leukocytes in homeostatic inflammation and in inflammatory disease. The change in title of the project from "Molecular Mechanisms of Neutrophil Adhesion to Endothelium" to "Molecular Interactions of Myeloid Cells with Endothelium" reflects a larger scope, including a focus on adhesion-dependent signaling mechanisms and new investigations of the interactions of monocytes with endothelium, in addition to studies of PMNs. Specific adhesion molecules tether endothelial cells and myeloid cells together; similarly, such tethering molecules can bind platelets to myeloid cells and can mediate adhesion between the myeloid leukocytes themselves. We will determine how these adhesive interactions lead to the generation of intracellular signals that alter critical inflammatory functions of the myeloid cells. We will focus on expression of immediate-early genes, on cell spreading, and on related cellular responses. We will determine how molecules known to tether myeloid leukocytes transmit signals, and orchestrate them when acting in concert with other factors. Using isolated myeloid leukocytes and cultured endothelium, transfected cells, model membranes and purified molecules, we will characterize the roles of selectins, in Specific Aims l and 2. In Specific Aim 3, we will use similar strategies to identify tethering and signaling mechanisms of ICAM-3, a new member of the ICAM family, in myeloid cell interactions. In Specific Aim 4, we will examine "outside-in" signaling by beta32 (CD11/CD18) integrins in myeloid cells using two traditional approaches, adhesion of leukocytes to specific ligands and incubation with antibodies that may induce functional alterations, and a novel approach - study of myeloid cells with targeted deletions of individual CD11/CD18 heterodimers. Information from these reduced systems will then be used to analyze complex models of inflammation (cell-cell interactions in vitro) and, ultimately, to identify new strategies for understanding and modifying inflammatory events in vivo and in humans.
Keywords: atherosclerosis, biological signal transduction, inflammation, leukocyte adhesion molecule, monocyte, neutrophil, CD antigen, cell cell interaction, dimer, gene expression, glycoprotein, homeostasis, integrin, membrane protein, molecular biology, protein purification, selectin, vascular endothelium, human tissue, laboratory rabbit, polymerase chain reaction, tissue /cell culture, transfection
Project start date: 1990-04-01
Project end date: 1999-03-31
5R01HL044525-09 (1998): $257553
5R01HL044525-08 (1997): $248047
2R01HL044525-06 (1995): $229656
Sponsored Links Excellgen http://Excellgen.com
MOLECULAR INTERACTIONS--MYELOID CELLS W ENDOTHELIUM
Guy A Zimmerman, Professor Of Internal Medicine
University Of Utah 75 South 2000 East Salt Lake City, Ut 84112
Grant 2R01HL044525-10 from National Heart, Lung, And Blood Institute IRG: PTHA
Abstract: The long term goal of this project is to understand how adhesive interactions induce and integrate intercellular signaling between monocytes and neutrophils, which are leukocytes of the myeloid lineage, and endothelial cells. We also will determine how these interactions influence subsequent functional responses of the leukocytes in homeostatic inflammation and in inflammatory disease. The change in title of the project from "Molecular Mechanisms of Neutrophil Adhesion to Endothelium" to "Molecular Interactions of Myeloid Cells with Endothelium" reflects a larger scope, including a focus on adhesion-dependent signaling mechanisms and new investigations of the interactions of monocytes with endothelium, in addition to studies of PMNs. Specific adhesion molecules tether endothelial cells and myeloid cells together; similarly, such tethering molecules can bind platelets to myeloid cells and can mediate adhesion between the myeloid leukocytes themselves. We will determine how these adhesive interactions lead to the generation of intracellular signals that alter critical inflammatory functions of the myeloid cells. We will focus on expression of immediate-early genes, on cell spreading, and on related cellular responses. We will determine how molecules known to tether myeloid leukocytes transmit signals, and orchestrate them when acting in concert with other factors. Using isolated myeloid leukocytes and cultured endothelium, transfected cells, model membranes and purified molecules, we will characterize the roles of selectins, in Specific Aims l and 2. In Specific Aim 3, we will use similar strategies to identify tethering and signaling mechanisms of ICAM-3, a new member of the ICAM family, in myeloid cell interactions. In Specific Aim 4, we will examine "outside-in" signaling by beta32 (CD11/CD18) integrins in myeloid cells using two traditional approaches, adhesion of leukocytes to specific ligands and incubation with antibodies that may induce functional alterations, and a novel approach - study of myeloid cells with targeted deletions of individual CD11/CD18 heterodimers. Information from these reduced systems will then be used to analyze complex models of inflammation (cell-cell interactions in vitro) and, ultimately, to identify new strategies for understanding and modifying inflammatory events in vivo and in humans.
Keywords: atherosclerosis, biological signal transduction, inflammation, leukocyte adhesion molecule, monocyte, neutrophil, CD antigen, cell cell interaction, dimer, gene expression, glycoprotein, homeostasis, integrin, membrane protein, molecular biology, protein purification, selectin, vascular endothelium, human tissue, laboratory rabbit, polymerase chain reaction, tissue /cell culture, transfection
Project start date: 1990-04-01
Project end date: 2004-03-31
2R01HL044525-10 (1999): $283308
MECHANISMS OF NEUTROPHIL ADHESION TO ENDOTHELIUM
Guy A Zimmerman, Professor Of Internal Medicine
University Of Utah 75 South 2000 East Salt Lake City, Ut 84112
Grant 5R01HL044525-05 from National Heart, Lung, And Blood Institute IRG: PTHA
Abstract: The goal of this project is to define the molecular mechanisms that regulate the adhesion of polymorphonuclear leukocytes (PMNs, neutrophils, granulocytes) to endothelial cells (EC) and the biologic consequences of this interaction, such as activation (and subsequent polarization, migration, degranulation, etc) of the PMNs. The regulation of PMN adhesion to the Ec that form the intima of blood vessels is a central issue in vascular biology because of its requisite role in physiologic inflammation and host defense. Furthermore, unregulated adhesion of PMNs to EC, followed by release of granular enzymes, oxygen radicals, and other mediators by the activated leukocytes, contributes to vascular damage in ischemia-reperfusion injury, myocardial infarction, acute pulmonary vascular injury, nad other syndromes in human disease. We have described a novel mechanism of EC-dependent adhesion of PMNs. The adhesive interaction occurs rapidly (within minutes) when cultured human EC are activated by an agonist and is mediated, at least in part, by the rapid, time-dependent, synthesis and surface expression by the EC of platelet-activating factor ("PAF"; 1-0-alkyl-2-acetyl-sn-glycero-3-phosphocholine), a unique glycero- phospholipid that interacts with PMNs and other cell types by binding to a surface receptor. This rapid EC-dependent mechanism of PMN adhesion is clearly different from that induced by cytokines, which occurs over hours, and likely is involved in the rapid localization of PMNs to specific areas of the intima that occurs in physiologic and pathologic inflammation. The current project has 4 specific objectives 1) We will define the requirement for an activation response by the PMNs for the adhesive interaction and the mechanisms that are involved. This also the first step in determining how adhesion of PMNs to activated EC influences subsequent events (such as polarization, release of enzymes and lipid mediators, etc by the PMNs). 2) We will define the role of the PMN CD11/CD18 adhesive glycoproteins, with the hypothesis that they play an amplifying rather than an obligate role. 3) We will characterize the role of other EC molecules in rapid EC- dependent adhesion, focusing initially on granule membrane protein 140 (GMP-140), a novel glycoprotein that is localized in secretory granules of the EC and that can be rapidly translocated to the plasma membrane. 4) We will define the mechanism(s) of PMN adhesion to EC activated by oxidants, a model relevant to inflammatory vascular injury in human disease. Based on the results of this line of investigation, we will be able to explore the mechanisms that lead to unregulated PMN adhesion and activation in other models of vascular injury and potentially, in vivo. The experiments will employ cultured EC, isolated native, fixed, and radiolabeled PMNs, neutrophil cytoplasts, myeloid cell lines, studies of signal transduction mechanisms in PMNs activated by agonists presented in the EC plasma membrane, in model membranes, or in the fluid phase, studies of surface expression of lipid (PAF) and glycoprotein (GMP-140) mediators by Ec, metabolic labeling of PAF and other products of activated EC and, potentially, isolation and characterization of new mediators expressed by EC or PMNs.
Keywords: atherosclerosis, cell adhesion, inflammation, neutrophil, cell cell interaction, glycoprotein, membrane protein, molecular biology, human tissue, laboratory rabbit, tissue /cell culture
Project start date: 1990-04-01
Project end date: 1995-03-31
5R01HL044525-05 (1994): $208778
5R01HL044525-04 (1993): $200411
5R01HL044525-03 (1992): $188166
Molecular Interactions Of Myeloid Cells With Endothelium
Guy A Zimmerman, Professor Of Internal Medicine
University Of Utah 75 South 2000 East Salt Lake City, Ut 84112
Grant 5R37HL044525-17 from National Heart, Lung, And Blood Institute IRG: PTHA
Abstract: The broad long-term goals of this project are to define mechanisms by which interactions between myeloid leukocytes, endothelial cells and platelets activate molecular pathways that regulate cellular function and phenotype, how these functions become dysregulated, and how dysregulated interactions and activities cause inflammatory vascular and tissue injury. Molecular mechanisms and pathways that control these events provide fundamental biologic insights; in addition, they are directly relevant to human disease, since dysregulated inflammatory cell-cell interactions are key features of major human syndromes that span a spectrum from the acute manifestations of sepsis to atherosclerosis and its complications. After initially characterizing key mechanisms of myeloid leukocyte tethering and adhesion, work in this project has focused on how outside-in signals delivered by these adhesion pathways and by inflammatory receptors lead to altered gene expression. In the last funding period we explored post-transcriptional mechanisms of gene regulation, an issue of evolving significance in human biology and disease in the post-genomic era, and have identified pathways for signal-dependent translation of critical messenger RNAs (mRNAs) in myeloid leukocytes, endothelial cells and platelets that were previously unrecognized and have physiologic significance. The current application builds on these discoveries, and will characterize novel signal dependent translation pathways and genes based on the central hypothesis that translational control and signal-dependent translation mechanisms regulate key innate immune cell phenotypes, functions and responses in acute and chronic inflammation. The specific aims are 1) to characterize inflammatory mechanisms that regulate the mTOR translational control pathway in myeloid leukocytes; 2) define new inflammatory activities of mTOR in specialized innate immune effector cells; 3) characterize novel signal dependent translation mechanisms in myeloid leukocytes; 4) test the hypothesis that translationally regulated genes in myeloid leukocytes comprise a sub-group of transcripts that code for critical inflammatory proteins and can be identified by their association with polyribosomes. The studies will resolve critical gaps in our knowledge of gene regulation in inflammatory systems, have direct clinical relevance, and are innovative because they explore new concepts and paradigms.
Keywords: biological signal transduction, cell cell interaction, genetic translation, inflammation, leukocyte, vascular endothelium, gene expression, immunogenetics, polysome, clinical research, human subject
Project start date: 1990-04-01
Project end date: 2009-03-31
5R37HL044525-17 (2006): $328471
5R37HL044525-16 (2005): $336375
Guy A Zimmerman
University Of Utah
Project start date: 1990-04-01
Project end date: 2014-01-31
MOLECULAR INTERACTIONS OF MYELOID CELLS WITH ENDOTHELIUM
Guy A Zimmerman
University Of Utah, 75 South 2000 East, Salt Lake City, Ut 84112
Grant 5R37HL044525-21 from National Heart, Lung, And Blood Institute
Keywords: Acute; Address; Adhesions; Atheroscleroses; Atherosclerosis; Atherosclerotic Cardiovascular Disease; Binding; Binding (Molecular Function); Bizzozero`s corpuscle/cell; Blood Platelets; Blood Vessels; Blood leukocyte; Body Tissues; Cell Communication; Cell Communication and Signaling; Cell Function; Cell Interaction; Cell Process; Cell Signaling; Cell model; Cell physiology; Cell-to-Cell Interaction; Cells; Cellular Function; Cellular Physiology; Cellular Process; Cellular model; Chronic; Code; Coding System; Deetjeen`s body; Disease; Disorder; Effector Cell; Endothelial Cells; Endothelium; Event; FK506 Binding Protein 12-Rapamycin Associated Protein 1; FK506 Binding Protein 12-Rapamycin Associated Protein 2; FK506 binding protein 12-rapamycin associated protein 1, human; FKBP-Rapamycin Associated Protein; FKBP-rapamycin associated protein, human; FKBP12 Rapamycin Complex Associated Protein 1; FRAP1 protein, human; Funding; Gene Action Regulation; Gene Expression; Gene Expression Regulation; Gene Regulation; Gene Regulation Process; Genes; Genomics; Goals; Hayem`s elementary corpuscle; Human; Human Biology; Human, General; INFLM; Immune; Inflammation; Inflammatory; Injury; Intracellular Communication and Signaling; Knowledge; Lead; Leukocytes; Man (Taxonomy); Man, Modern; Marrow leukocyte; Marrow platelet; Messenger RNA; Molecular; Molecular Interaction; Myelogenous; Myeloid; Myeloid Cells; Pathway interactions; Pb element; Phenotype; Physiologic; Physiological; Platelets; Polyribosomes; Polysomes; Productivity; Programs (PT); Programs [Publication Type]; Progress Reports; Proteins; RAFT1 protein, human; RAPT1 protein, human; RNA, Messenger; Rapamycin Target Protein; Receptor Protein; Reports, Progress; Reticuloendothelial System, Leukocytes; Reticuloendothelial System, Platelets; Role; SUBGP; Sepsis; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Subcellular Process; Subgroup; Syndrome; System; System, LOINC Axis 4; Testing; Thrombocytes; Tissues; Transcript; Translations; White Blood Cells; White Cell; Work; atheromatosis; atherosclerotic vascular disease; base; biological signal transduction; bloodstream infection; clinical relevance; clinically relevant; disease/disorder; gene product; heavy metal Pb; heavy metal lead; human FRAP1 protein; human disease; innovate; innovation; innovative; insight; mRNA; mTOR; novel; pathway; programs; rapamycin and FKBP12 target 1 protein, human; receptor; response; social role; thrombocyte/platelet; vascular; white blood cell; white blood corpuscle
Project start date: 1990-04-01
Project end date: 2014-01-31
Budget start date: 1-APR-2010
Budget end date: 31-JAN-2011
5R37HL044525-21 (2010): $363781
Sponsored Links Excellgen http://Excellgen.com
A Murine Model For Inhibition Of Farnesyltransferase
Guy A Zimmerman, Professor Of Internal Medicine
Human Geneticsuniversity Of Utah
Grant 5R03NS048512-02 from National Institute Of Neurological Disorders And Stroke IRG: AICS
Project start date: 2008-02-01
Project end date: 2009-12-31
Molecular Interactions Of Myeloid Cells With Endothelium
Guy A Zimmerman, Professor Of Internal Medicine
Cardiovascular Research And Training Instituteuniversity Of Utah
Grant 4R37HL044525-20 from National Heart, Lung, And Blood Institute IRG: NSS
Project start date: 1990-04-01
Project end date: 2014-03-31
5R37HL044525-18 (2007): $318945
2R01HL044525-15 (2004): $336375
5R01HL044525-07 (1996): $238587