# projects

application_id: 8597407
project_number: 5I01BX001014-04
project_title: Regulation of adipose cells by autotaxin / lysophosphatidic acid signaling
administering_ic: VA
award_notice_date: 2013-11-06
opportunity_number: BX-10-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 001014
project_start_date: 2010-10-01
project_end_date: 2014-09-30
study_section: Endocriniology A[ENDA]
contact_pi_person_id: 1932626
contact_pi: SMYTH, SUSAN S.
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2013-10-01
project_abstract:
DESCRIPTION (provided by applicant):     Project Summary/Abstract Obesity and obesity-related cardiovascular complications are an increasing health burden in the United States. An analysis of nearly 2 million veterans receiving care at VA Medical Centers in 2000 indicated that ~68% were overweight and ~37% were obese. Despite extensive investigation viable therapeutic strategies to prevent weight gain or promote weight loss remain largely elusive. Attention has recently focused on the exciting possibility that a therapeutic strategy aimed at promoting energy expenditure by increasing brown adipose tissue (BAT) content or activity in adults could reduce fat deposition and obesity. However such a therapeutic strategy requires a comprehensive understanding of factors that regulate BAT. Lysophosphatidic acid (LPA) is a bioactive lipid present in physiologically relevant levels in plasma and other biologic fluids and is positioned to play a role in human health and disease. The secreted lysophospholipase D autotaxin (autotaxin/lysoLPD) is responsible for production of extracellular biologically active LPA which is in turn inactivated by enxymatic dephosphorylation by lipid phosphate phosphatases (LPP). We provide evidence that treatment of primary cultures of preadipocytes with LPA or autotaxin reduces expression of markers of brown adipocyte differentiation and LPP3 an endogenous regulator of Wnt; whereas a novel autotaxin/lysoPLD inhibitor dramatically promotes brown adipocyte differentiation. In mice overexpression of autotaxin reduces UCP1 expression and promotes diet-induced obesity. We therefore propose the central hypothesis that signaling pathways regulated by autotaxin and LPP3 contribute to diet-induced thermogenesis and regulate the development of diet-induced obesity. We will test our central hypothesis using state-of-the art genetic and pharmacologic approaches in three specific aims. In Aim One we will identify the role of autotaxin/lysoPLD in regulation of brown adipose tissue and diet-induced obesity in animal models. In Aim Two we will establish the molecular mechanism(s) by which LPA and autotaxin exert their effects. In Aim Three we identify the role of LPP3 in development and function of adipose tissue and diet-induced obesity. The aims of this grant provide a vehicle to address a major unresolved issue in the field of lysolipid signaling namely role of autotaxin/lysoPLD LPA and LPP3 in regulation of adiposity. These results will be significant because they are expected to provide innovative targets and provide proof-of-concept for novel inhibitors that may be used for prevention and treatment of obesity in humans.    PUBLIC HEALTH RELEVANCE:     Project Narrative More than 44 million Americans are obese as defined by body mass index (BMI) of >30 kg/m2. Obesity is a significant health care problem among veterans receiving care at VA Medical Centers. A comprehensive analysis of nearly 2 million veterans in 2000 indicated that ~68% were overweight and ~37% were obese. Current strategies to treat and prevent obesity focus primarily on reducing energy consumption through appetite suppression and behavioral modification. Recent attention has focused on an attractive alternative approach namely that of increasing energy expenditure by enhancing the heat-dissipating function of brown adipose tissue in adults. We provide evidence that the secreted lysophospholipase D autotaxin responsible for production of biologically active LPA may regulate the functional status of brown adipose tissue. The goal of the current study is to identify the role of the autotaxin/LPA signaling nexus in the regulation of adipose tissue and diet-induced obesity. Our results may suggest innovative targets to prevent and treat obesity in humans.

project_terms:
Address;Adipocytes;Adipose tissue;Adult;Affect;American;Animal Model;Attention;Attenuated;Behavior Therapy;Body Weight decreased;Body mass index;Brown Fat;Cardiovascular system;Caring;Cell surface;Consumption;Data;Deposition;Desire for food;Development;Diet;Disease;Down-Regulation;Energy Metabolism;Exhibits;Family;Fatty acid glycerol esters;G-Protein-Coupled Receptors;Gap Junctions;Generations;Genetic;Goals;Grant;Health;Healthcare;Heating;Human;In Vitro;Integral Membrane Protein;Investigation;Knowledge;Lipids;Liquid substance;Lysophospholipids;Medical center;Molecular;Mus;Names;Obesity;Organism;Overweight;Plasma;Play;Positioning Attribute;Prevention;Production;Protein Dephosphorylation;Reagent;Recombinants;Regulation;Reporting;Research;Role;Signal Pathway;Signal Transduction;System;Techniques;Testing;Therapeutic;Thermogenesis;Tissue Differentiation;United States;Veterans;Weight Gain;Work;abstracting;adipocyte biology;adipocyte differentiation;alkylglycerophosphoethanolamine phosphodiesterase;extracellular;functional status;improved;in vivo;inhibitor/antagonist;innovation;lipid phosphate phosphatase;lysophosphatidic acid;member;novel;obesity prevention;obesity treatment;overexpression;precursor cell;prevent;public health relevance;tool;transdifferentiation


application_id: 8597360
project_number: 5I01BX000693-04
project_title: Airway sensors with myelinated afferents
administering_ic: VA
award_notice_date: 2013-11-08
opportunity_number: BX-10-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000693
project_start_date: 2010-10-01
project_end_date: 2014-09-30
study_section: Respiration[PULM]
contact_pi_person_id: 2084836
contact_pi: YU, JERRY (JUN)
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2013-10-01
project_abstract:
DESCRIPTION (provided by applicant):     The long-term objective of this proposal is to identify the unknown morphology and pathophysiological functions of pulmonary sensory receptors potentially benefiting more than 30 million patients with chronic obstructive pulmonary disease (COPD) and other pulmonary diseases in the US including veterans. Dyspnea bronchial inflammation hyperreactivity coughing and mucus secretion occur when airway sensory receptors are activated. These are common clinical manifestations of pulmonary disorders; however the underlying neural mechanisms remain unclear in part because our knowledge of airway receptor morphology and sensory behavior is limited. Increasing evidence points to important roles of myelinated airway receptors in pulmonary diseases. In this application we will test the hypothesis that vagal airway sensory units are not only transducers but also processors. Sensory units' behaviors depend on their structures. In addition a single unit may contain homogeneous or heterogeneous types of receptors providing varied mixed behavior. In this application we will address three specific aims to characterize the morphology and sensory behavior of myelinated airway receptors. 1) To identify different morphological structures of myelinated airway receptors by examing receptor morphology by immunohistochemical or neural tracing techniques combined with confocal microscopy. 2) To assign a specific receptor structure to physiologically defined classes of receptors by first electrophysiologically characterizing each known receptor type then tissue in the receptive field will be isolated dissected and immunohistochemically stained for receptor morphology. 3) To explore mechanosensor structure-function relationships and sensory integration mechansims by examining sensory behavior before and after blocking or stimulating one of the receptors in the unit. Thus integration mechanisms of sensory units can be explored. Successful completion of these studies will provide important structure-function relationships and establish a comprehensive objective classification system for the airway receptors. The presence of heterogenous receptors challenges the current view of the airway sensory unit. Transmission of heterogeneous information in a single unit verifies that the unit is not merely a transducer but also a processor. Our data will lay the foundation for future research and facilitate our understanding of the receptor function and their roles in pulmonary diseases and may help improve managements for cardiopulmonary patients suffering from cardiac failure COPD asthma acute lung injury and other pulmonary diseases.    PUBLIC HEALTH RELEVANCE:     Airway sensory receptors play important roles in cardiopulmonary diseases. Understanding their mechanisms of operation may help patient management thus benefitting a large number of veterans who suffer from cardiac and lung diseases including cardiac failure acute respiratory distress syndrome (ARDS) chronic obstructive pulmonary disease (COPD) and asthma.

project_terms:
Address;Adult Respiratory Distress Syndrome;Asthma;Behavior;Bronchoconstriction;C Fiber;Cardiopulmonary;Chemicals;Chronic Obstructive Airway Disease;Classification;Clinical;Confocal Microscopy;Cor pulmonale;Coughing;Data;Development;Disease;Dyspnea;Exhibits;Fiber;Foundations;Heart Diseases;Heart failure;Inflammation;Knowledge;Laboratories;Lung;Lung Injury Acute;Lung diseases;Mechanics;Mechanoreceptors;Microscopy;Morphology;Mucous body substance;Neurons;Patients;Physiological;Play;Process;Research;Role;Sensory;Sensory Receptors;Staining method;Stains;Structure;Structure of parenchyma of lung;Structure-Activity Relationship;Surveys;System;Techniques;Testing;Tissues;Trachea;Transducers;Vagus nerve structure;Veterans;improved;neuromechanism;operation;public health relevance;pulmonary function;receptive field;receptor;receptor function;relating to nervous system;response;sensor;sensory integration;sensory mechanism;transmission process

public_health_relevance:
Airway sensory receptors play important roles in cardiopulmonary diseases. Understanding their mechanisms of operation may help patient management thus benefitting a large number of veterans who suffer from cardiac and lung diseases including cardiac failure acute respiratory distress syndrome (ARDS) chronic obstructive pulmonary disease (COPD) and asthma.


application_id: 8586860
project_number: 5I01CX000240-04
project_title: Mechanisms of hepcidin effects in the anemia of chronic disease
administering_ic: VA
award_notice_date: 2013-11-08
opportunity_number: CX-10-001
application_type: 5
activity_code: I01
ic: CX
serial_number: 000240
project_start_date: 2010-10-01
project_end_date: 2014-09-30
study_section: Hematology[HEMA]
contact_pi_person_id: 1876507
contact_pi: MEANS, ROBERT T
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2013-10-01
project_abstract:
DESCRIPTION (provided by applicant):     The anemia of chronic disease (ACD) is one of the most common clinical syndromes encountered in the practice of medicine. This disorder typically manifests itself as a hypoproliferative anemia accompanied by a low serum iron concentration despite adequate reticuloendothelial iron stores. ACD has generally been considered to result from a combination of pathologic processes which can be linked to the cytokine mediators of inflammation. However the diagnostic importance of altered iron metabolism in ACD has led many investigators to consider that this feature denotes the dominant pathophysiologic mechanism of this syndrome. Hepcidin is an antibacterial protein which is produced in the liver circulates in the blood and is excreted in the urine. It is a type II acute phase protein the expression of which is induced by interleukin (IL)-6 and down-regulated by tumor necrosis factor (TNF). The specific iron regulatory functions of hepcidin as well as the anemia syndromes observed in patients and transgenic animals with abnormalities of hepcidin gene expression (discussed below) have led many to consider that it is the key to unlocking "the mysteries of ACD ".  Three major processes are involved in the pathogenesis of ACD. A slight shortening of red cell survival creates a demand for a small increase in red cell production by the bone marrow. The marrow cannot respond adequately to this demand due to impaired erythropoiesis and impaired mobilization of reticuloendothelial system iron stores. A potential role for hepcidin in the iron anomalies of ACD is strongly supported by available data. However iron changes are not the sole abnormality characteristic of ACD. If hepcidin is the major factor responsible for ACD then it should also contribute to the impaired erythropoiesis observed in this syndrome.  The hypothesis that hepcidin contributes to impaired erythropoiesis in ACD will be addressed through the following specific aims: 1. Determination of hepcidin effects on erythroid colony formation and investigation of the mechanisms involved. This will include A. whether the effects of hepcidin on erythropoiesis are direct-acting or require an accessory cell; B. the role of Epo concentration in hepcidin effects on erythropoiesis; C. the role of apoptosis/anti-proliferation in inhibitory effects of hepcidin on erythropoiesis; D. the effects of cytokines on the hepcidin/erythropoiesis relationship; E. the contributions of iron to hepcidin effects on CFU-E colony formation; F. the role of ferroportin (FPN) and variant FPN in hepcidin effects on CFU-E; and G. effects of Epo and inflammatory cytokines on FPN variant expression in erythroid cells. 2. Determination of the interactions of hepcidin and cytokines in bone marrow macrophage iron transport. Hepcidin inhibits egress of iron from bone marrow macrophages by downregulation of the iron export protein FPN. The ability of inflammatory cytokines to enhance or diminish this effect will be determined as well as any effects on the expression of variant FPN.3. Determination of hepcidin effects on induced Epo production in vitro and delineation of involved mechanisms. This will include A. determination and quantification of the effects of hepcidin on the induction of Epo protein in hepatic and renal models and of the time-course of these effects; and B. determination of the roles of inflammatory cytokines in modulating these effects; C. the role of iron availability in these effects; and D. determination of the effects of hepcidin on Epo transcription and the mechanism involved in these effects. 4. Effects of rhEpo on hepcidin production in vitro. In this Aim we will determine A. the effects of Epo on hepcidin production by HepG2 as well as B. the extent to which these effects are modulated by TNF IL-1 and IL-6. Relevance to VA mission: The proposed studies can potentially lead to an enhanced understanding of the mechanisms of ACD and lead to more effective therapy.    PUBLIC HEALTH RELEVANCE:     Anemia is a major contributor to morbidity and mortality in chronically ill patients and the anemia of chronic disease (ACD) is the major etiology of this anemia. This disorder typically manifests itself as a hypoproliferative anemia accompanied by a low serum iron concentration despite adequate reticuloendothelial iron stores. ACD is highly prevalent in the VA population. In ACD a slight shortening of red cell survival creates a demand for a small increase in red cell production by the bone marrow. The marrow cannot respond adequately to this demand due to impaired erythropoiesis and impaired mobilization of reticuloendothelial system iron stores. A potential role for hepcidin in the iron anomalies of ACD is strongly supported by available data. If hepcidin is the major factor responsible for ACD then it should also contribute to the impaired erythropoiesis observed in this syndrome. The proposed studies can potentially lead to an enhanced understanding of the mechanisms of ACD and therefore to more effective therapy.

project_terms:
Acute-Phase Proteins;Address;Anemia;Anemia due to Chronic Disorder;Anti-Bacterial Agents;Antigen-Presenting Cells;Apoptosis;Blood;Bone Marrow;CFU-E;Cell Survival;Characteristics;Chronically Ill;Clinical;Data;Diagnostic;Disease;Down-Regulation;Erythrocytes;Erythroid;Erythroid Cells;Erythropoiesis;Etiology;Gene Expression;Genetic Transcription;Hepatic;In Vitro;Inflammation Mediators;Inflammatory;Interleukin-1;Interleukin-6;Investigation;Iron;Kidney;Lead;Link;Liver;Marrow;Medicine;Mission;Modeling;Morbidity - disease rate;Mortality Vital Statistics;Pathogenesis;Pathologic Processes;Patients;Population;Process;Production;Protein Export Pathway;Proteins;Research Personnel;Reticuloendothelial System;Role;Serum;Syndrome;Time;Transgenic Animals;Tumor Necrosis Factor-alpha;Urine;Variant;cytokine;effective therapy;hepcidin;iron metabolism;macrophage;metal transporting protein 1;protein expression;public health relevance

public_health_relevance:
Anemia is a major contributor to morbidity and mortality in chronically ill patients and the anemia of chronic disease (ACD) is the major etiology of this anemia. This disorder typically manifests itself as a hypoproliferative anemia accompanied by a low serum iron concentration despite adequate reticuloendothelial iron stores. ACD is highly prevalent in the VA population. In ACD a slight shortening of red cell survival creates a demand for a small increase in red cell production by the bone marrow. The marrow cannot respond adequately to this demand due to impaired erythropoiesis and impaired mobilization of reticuloendothelial system iron stores. A potential role for hepcidin in the iron anomalies of ACD is strongly supported by available data. If hepcidin is the major factor responsible for ACD then it should also contribute to the impaired erythropoiesis observed in this syndrome. The proposed studies can potentially lead to an enhanced understanding of the mechanisms of ACD and therefore to more effective therapy.


application_id: 8443030
project_number: 1I01BX001984-01
project_title: Association of a common variant of the PPAP2B gene with cardiovascular disease.
administering_ic: VA
award_notice_date: 2013-03-01
opportunity_number: BX-12-001
application_type: 1
activity_code: I01
ic: BX
serial_number: 001984
project_start_date: 2013-01-01
project_end_date: 2016-12-31
study_section: Cellular and Molecular Medicine[CAMM]
contact_pi_person_id: 1879477
contact_pi: MORRIS, ANDREW J
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2013-01-01
project_abstract:
DESCRIPTION (provided by applicant):   
   
Ischemic heart disease caused by atherosclerosis of the coronary arteries remains the most frequent cause of mortality among Veterans despite extensive investigation into its pathobiology the identification of many risk factors and development of new therapeutic strategies. The hallmark features of endothelial damage lipid deposition smooth muscle cell proliferation and vascular inflammation contribute to the development and complications of atherosclerosis. The bioactive lysophospholipids sphingosphine-1-phosphate (S1P) and lysophosphatidic acid (LPA) act on cell surface receptors expressed by many vascular cells. These lipids are positioned to serve as mediators of the cellular events contributing to atherosclerosis and their role is supported by emerging evidence from experimental models. The lipid phosphate phosphatase 3 (encoded by the PPAP2B gene) is a cell surface integral membrane protein that regulates the bioavailability of S1P and LPA by catalyzing their dephosphorylation to generate lipid products that are not receptor active. Analysis of data from a series of genome-wide association studies (GWAS) focuses attention on a striking association between a single nucleotide polymorphism (SNP) in the PPAP2B locus and coronary artery disease. This rs17114036 SNP lies in an intronic non-coding region of the gene that could affect gene expression. How genetic variation in PPAP2B confers risk of coronary artery disease is unknown in large part because of a lack of understanding of LPP3 function in vascular cells. In this application we present evidence that vascular cell LPP3 serves as an intrinsic negative regulator of vascular inflammation suppresses smooth muscle cell proliferation and promotes endothelial barrier function. These protective effects of LPP3 suggest that PPAP2B polymorphisms associated with reduced gene expression could aggravate cellular events underlying atherosclerosis and increase the likelihood of myocardial infarction.  The broad long-term goal of this research project is to explain how human genetic variation at the PPAP2B locus alters the risk of myocardial infarction and apply that knowledge to improve the diagnosis and therapy of ischemic heart disease. We have assembled a team of experts whose knowledge spans all aspects of the proposed research to position ourselves to validate the function of the PPAP2B locus in atherosclerosis. In this proposal we will test our central hypothesis which is that the minor allele of rs17114036 in PPAP2B reduces gene expression and that lower levels of LPP3 promote atherosclerosis. We will apply unique tools that we have developed to study LPP3 and our considerable expertise to test our central hypothesis. Firstly we will identify the mechanism by which in the rs17114036 SNP in PPAP2B affects LPP3 expression and activity. We predict that rs17114036 affects a U1 splisome binding site and thereby reduces gene expression.  We will test this working hypothesis by determining if the minor allele of rs17114036 associates with altered  LPP3 transcript and protein levels in white blood cells and arterial tissue and by examining the consequences of the polymorphism on RNA splicing and stability. Secondly we will establish a mechanistic role for LPP3 in experimental atherosclerosis. Based on our Preliminary Studies of vascular pathology in mice with tissue-specific defects in smooth muscle and endothelial LPP3 the working hypothesis of this section is that LPP3 protects against vascular inflammation and that reduced expression will therefore accelerate atherosclerosis.  We will test this working hypothesis by determining the consequences of vascular tissue specific deficiency of LPP3 on the development of atherosclerosis in mice.  The experiments proposed in this application are an important step in functionally validating a common genetic variant in PPAP2B as a cardiovascular risk predictor in humans. Establishing PPAP2B its product LPP3 and the lipid substrates of this enzyme as risk predictors and mediators of CAD promises to identify important and innovative targets for the development of new biomarkers and/or therapeutics.

project_terms:
Accounting;Address;Admission activity;Affect;Alleles;Atherosclerosis;Attention;Binding Sites;Biological Availability;Biological Markers;Blood Pressure;Blood Vessels;Cardiovascular Diseases;Cardiovascular system;Cell Proliferation;Cell Surface Receptors;Cell surface;Cells;Cholesterol;Clinical;Congestive Heart Failure;Coronary Arteriosclerosis;Coronary artery;Data Analyses;Defect;Deposition;Development;Diabetes Mellitus;Diagnosis;Disease;Disease susceptibility;Employee Strikes;Enzymes;Event;Experimental Models;Family;Frequencies (time pattern);Functional RNA;Gene Expression;Genes;Genetic Polymorphism;Genetic screening method;Genotype;Goals;Health;Healthcare;Hospitals;Human;Human Genetics;Hypertension;Individual;Integral Membrane Protein;Investigation;Knowledge;LDL Cholesterol Lipoproteins;Lead;Leukocytes;Lipids;Lysophospholipids;Mediator of activation protein;Membrane;Messenger RNA;Minor;Mortality Vital Statistics;Mus;Muscle Smooth Vascular;Mutation;Myocardial Infarction;Myocardial Ischemia;Nuclear;Pathology;Plasma;Positioning Attribute;Predisposition;Prevention;Process;Protein Dephosphorylation;Proteins;RNA Splicing;RNA Stability;Reagent;Recording of previous events;Research;Research Project Grants;Resolution;Ribonucleoproteins;Risk;Risk Factors;Role;Sampling;Series;Single Nucleotide Polymorphism;Smooth Muscle Myocytes;Smooth muscle (tissue);Specific qualifier value;Testing;Therapeutic;Tissues;Transcript;Translating;United States;Validation;Variant;Variation (Genetics);Vascular Endothelial Cell;Veterans;Work;base;cardiovascular disorder risk;cardiovascular risk factor;disorder risk;enzyme activity;enzyme substrate;genetic variant;genome wide association study;improved;innovation;inorganic phosphate;lipid metabolism;lipid phosphate phosphatase;lysophosphatidic acid;novel strategies;novel therapeutics;protective effect;public health relevance;receptor;research study;tool;vascular inflammation

public_health_relevance:
Cardiovascular diseases are one of the most prevalent health problems among Veterans resulting in over 25000 admissions to VA hospitals annually. Family history is well established to provide a strong clinical indicator of cardiovascular disease risk but until very recently in al but a few cases the genes involved were not known. A new approach called "genome wide association" has been used to identify common genetic variants that predict cardiovascular disease risk without obvious association with established cardiovascular disease risk factors such as elevated plasma cholesterol levels and increased blood pressure. The goal of this project is to determine why a common variant in a gene called PPAP2B is associated with significantly elevated cardiovascular disease risk. Completion of the work could lead to validation of this variant as a genetic test for cardiovascular disease susceptibility and the development of new therapies for treatment and prevention of cardiovascular to enable improved and more efficient healthcare for Veterans.


application_id: 8442018
project_number: 1I01BX001838-01A1
project_title: Mechanism of Neutrophil Activation
administering_ic: VA
award_notice_date: 2013-02-15
opportunity_number: BX-12-001
application_type: 1
activity_code: I01
ic: BX
serial_number: 001838
suffix: A1
project_start_date: 2012-10-01
project_end_date: 2016-09-30
study_section: Immunology and Dermatology A[IMMA]
contact_pi_person_id: 1886516
contact_pi: MCLEISH, KENNETH R
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
project_abstract:
DESCRIPTION (provided by applicant):    
   
Neutrophils are a critical component of both the innate and adaptive immune responses that control microbial invasion. Priming is an intermediate step by which resting neutrophils enhance their production of reactive oxygen intermediates (ROI) upon activation and priming is necessary for optimal microbial killing. Although priming greatly enhances neutrophil bactericidal capability primed neutrophils are also more dangerous to normal cells. Primed neutrophils participate in a number of diseases prominent in the aging Veteran population including ANCA-associated vasculitis ischemia- reperfusion injury acute respiratory distress syndrome rheumatoid arthritis peridontitis atherosclerosis and acute inflammatory liver injury. Thus manipulation of neutrophil priming is a potential therapeutic strategy for pharmacological intervention to restrain inflammation. Recent studies identified granule exocytosis and enhanced translocation of cytosolic components of the NADPH oxidase to the plasma membrane as possible mechanisms of neutrophil priming. However many gaps in knowledge remain. The relative contribution of each of those events to priming has not been examined and whether both are necessary or sufficient for priming is unknown. The molecular events by which granule exocytosis contributes to priming and the molecular events that control enhanced translocation of cytosolic NADPH oxidase components are incompletely understood. Finally the ability to manipulate neutrophil priming in vivo has not been established. The McLeish laboratory is in a unique position to address each of these gaps based on our extensive experience examining signal transduction pathways our application of biochemical genetic and proteomic approaches to understand neutrophil biology and our recent development of novel reagents that inhibit exocytosis in vitro and in vivo. The current proposal will address the central hypothesis that neutrophil priming results from the convergence of signal transduction pathways into two responses an exocytose-dependent increase in plasma membrane expression of membrane components of NADPH oxidase and prolyl isomerase-dependent conformational changes in cytosolic components of NADPH oxidase. The following four specific objectives will be accomplished.  Specific Objective 1 will test the working hypothesis that exocytosis of secretory vesicles and gelatinase granules results in increased plasma membrane expression of p91phox and p22phox which is necessary but not sufficient for priming. Specific Objective 2 will test the working hypothesis that activation of the prolyl isomerase Pin1 contributes to priming of human neutrophils by multiple mechanisms including enhanced p47phox translocation and increased granule exocytosis. Specific Objective 3 will test the working hypothesis that p38 MAPK phosphorylates multiple proteins that mediate events necessary for priming including exocytosis and Pin1 activation. Specific Objective 4 will test the
working hypothesis that neutrophil priming is a necessary component for neutrophil participation in inflammatory diseases and represents a viable therapeutic strategy using an animal model of acute lung injury (ALI). The proposed work is expected to establish the molecular basis for neutrophil priming. The impact of the project is that molecular targets identified will permit therapeutic manipulation of neutrophil priming in inflammatory diseases. Thus new approaches to treating Veteran patients with a number of acute and chronic inflammatory diseases are anticipated.

project_terms:
Acute;Address;Adult Respiratory Distress Syndrome;Affect;Aging;Animal Model;Antineutrophil Cytoplasmic Antibodies;Arteriosclerotic Cardiovascular Disease;Atherosclerosis;Bacteria;Biochemical Genetics;Biology;Cell membrane;Chemicals;Chronic;Cytoplasmic Granules;Data;Development;Disease;Event;Exocytosis;Exposure to;Gelatinases;Goals;Human;Immune response;In Vitro;Infection;Infection Control;Inflammation;Inflammation Mediators;Inflammatory;Injury;Intervention;Invaded;Killings;Knowledge;Laboratories;Lead;Leukocytes;Lung Injury Acute;MAP Kinase Gene;MAPK14 gene;Mediating;Membrane;Molecular;Molecular Target;NADPH Oxidase;Neutrophil Activation;Normal Cell;Oxidases;Patients;Peptidylprolyl Isomerase;Phosphorylation;Play;Poisons;Population;Positioning Attribute;Process;Production;Proteins;Proteomics;Reagent;Relative (related person);Reperfusion Injury;Reporting;Rest;Rheumatoid Arthritis;Role;Secretory Vesicles;Signal Transduction Pathway;Testing;Therapeutic;Tissues;Toxin;Vasculitis;Veterans;Work;bactericide;base;experience;extracellular;human CYBA protein;human disease;in vivo;liver injury;lung vascular injury;microbial;neutrophil;novel;novel strategies;public health relevance;reactive oxygen intermediate;response;sound;therapeutic target;tool

public_health_relevance:
White blood cells (WBCs) protect against infection by producing toxic chemicals that kill invading bacteria. The production of those toxin chemicals is enhanced by prior exposure to a variety of mediators of inflammation by a process termed priming. Although WBC priming plays a critical role in the control of infection priming is a two-edged sword that also reduces the ability of WBCs to co-exist with normal cells. Primed WBCs produce tissue injury in a number of diseases that affect our Veteran patients. The events that lead to priming are not known and inhibition of priming has not been used to treat those diseases. The current proposal will define the mechanisms by which WBCs become primed and will establish priming is a therapeutic target in inflammatory diseases. Accomplishing those goals will provide new treatments for Veteran patients with a number of acute and chronic inflammatory diseases.


application_id: 8398935
project_number: 5I01BX000962-03
project_title: Exosomal TRAF2-CSN5 complex mediated inflammation promotes tumor growth
administering_ic: VA
award_notice_date: 2013-07-04
opportunity_number: BX-10-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000962
project_start_date: 2011-04-01
project_end_date: 2015-03-31
study_section: Immunology and Dermatology A[IMMA]
contact_pi_person_id: 1889248
contact_pi: ZHANG, HUANG-GE
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2013-04-01
project_abstract:
DESCRIPTION (provided by applicant):     The overall goal of this proposal is to elucidate the mechanisms by which lung tumor exosomes promote an inflammatory response namely the activation of macrophages and promote the progression and growth of lung tumors. Our preliminary data indicate that pretreatment of A/J mice with exosomes produced by TC-1 lung tumor cells or exosomes isolated from the lung tissue of A/J mice that have been pretreated with the carcinogen urethane results in more rapid tumor growth and earlier progression of lung carcinomas. Urethane treatment results in the recruitment of the activated form of TRAF2 to the exosomes and the exosomes with activated TRAF2 are taken up by the bone marrow-derived precursors of macrophages leading to their maturation and subsequent migration to the lung. The recruitment of the activated TRAF2 to the exosomes requires degradation of the inflammation suppressor protein CYLD in the tumor cells; moreover TRAF2 can mediate ubiquination of CYLD thereby promoting its degradation. The results of siRNA TRAF2 knockout indicated however that TRAF2 is required but is not sufficient for ubiquitination of CYLD. We have now identified two exosomal proteins that interact with TRAF2 Itch and CSN5 which have the potential to enhance the ubiquitination of CYLD in lung tumor cells. Our recent data indicating that CYLD is not packed in exosomes isolated from the peripheral blood of lung cancer patients although it is present in exosomes from healthy volunteers indicate the potential clinical relevance of this model that directly links the effects of a carcinogen with an inflammatory response and promotion of tumor development. We propose to: (1) Confirm the role of lung tumor exosomal TRAF2 in the promotion of urethane-induced lung carcinomas by determining whether knockout of TRAF2 in TC-1 lung tumor cells is sufficient for (i) inhibition of macrophage differentiation and (ii) prevention of lung tumor exosome-mediated enhancement of urethane induced lung cancer. (2) Determine if other exosomal proteins that interact with CYLD participate in the ubiquitination of CYLD in TC-1 tumor cells and determine if the elimination of these proteins stabilizes exosomal CYLD and suppresses exosomal TRAF2 activation resulting in the inhibition of macrophage activation and prevention of tumor growth. (3) Determine if macrophages pulsed with lung tumor exosomes isolated from lung cancer patients promote lung tumor growth in a NOD-SCID mouse model. The data generated should permit the development of a highly innovative model of the cellular and molecular basis for exosome-mediated chronic inflammation and promotion of tumor growth and indicate novel preventive and therapeutic strategies.

project_terms:
A/J Mouse;Acceleration;Anti-Inflammatory Agents;Bone Marrow;Bone Marrow Cells;Cancer Patient;Carcinogens;Cell model;Cells;Chronic;Complex;Curcumin;Data;Development;Epithelial;Epithelial Cells;Goals;Growth;Human;IL8 gene;ITGAM gene;Inflammation;Inflammatory Response;Interleukin-6;Knock-out;Lead;Link;Lung;Lung Neoplasms;Macrophage Activation;Malignant neoplasm of lung;Mediating;Membrane;Modeling;Molecular;Mus;Myelogenous;Myeloid Cells;NOD/SCID mouse;Physiologic pulse;Play;Prevention;Preventive;Production;Property;Proteins;Research;Role;SCID Mice;Small Interfering RNA;Structure of parenchyma of lung;TRAF2 gene;Testing;Therapeutic;Tissues;Tumor Necrosis Factor-alpha;Tumor Promotion;Tumor Tissue;Tumor-Derived;Ubiquitination;Urethane;Vascular Endothelial Growth Factors;Vesicle;base;cancer cell;cell growth;clinically relevant;healthy volunteer;human TNF protein;in vivo;innovation;lung Carcinoma;macrophage;migration;mouse model;neoplastic cell;novel;peripheral blood;precursor cell;public health relevance;tumor;tumor growth;ubiquilin

public_health_relevance:
The proposed research is highly relevant to characterizing the association of lung tumor exosomes mediated inflammatory responses and lung tumor growth in that it is focused on characterizing the molecular mechanisms underlying the ability of tumor exosomes to both activate macrophages and promote tumor growth. The central hypothesis is that exosomes produced by lung tumor cells or lung tumor tissue are taken up by bone marrow myeloid precursor cells. These cells further differentiate into macrophages and migrate into the lung promote the progression of urethane- induced lung cancer.


application_id: 8394611
project_number: 5I01BX000387-04
project_title: Mood-Stabilizing Medications and the Inositol Signaling System
administering_ic: VA
award_notice_date: 2013-07-03
opportunity_number: BX-09-008
application_type: 5
activity_code: I01
ic: BX
serial_number: 000387
project_start_date: 2010-04-01
project_end_date: 2014-03-31
study_section: Neurobiology A[NURA]
contact_pi_person_id: 7118863
contact_pi: PARTHASARATHY, RANGA N
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2013-04-01
project_abstract:
DESCRIPTION (provided by applicant):     Project Summary/Abstract Bipolar disorder (BD) is a devastating condition affecting many Veterans. Lithium and valproate are the drugs of choice for the manic phase of BD. While therapeutically effective their mechanisms of action are not fully understood and undesirable side effects are common. Lithium inhibits brain inositol monophosphatase (IMPase) thereby depleting the available pool of free inositol and resulting in the correction of the manic phase of BD. Inositol synthase (IPS) is a key enzyme that regulates inositol levels by generating the precursor inositol phosphate (IP). Our preliminary data demonstrate that rat brain IPS is up-regulated by chronic lithium treatment which would have the same net effect as inositol depletion relative to inositol phosphate. We hypothesize that lithium and other mood stabilizers exert their therapeutic effects by up-regulating IPS expression increasing the available inositol phosphate pool. This implies that the pathophysiological mechanism underlying BD is a disturbance in maintaining a proper balance in the ratio of free inositol to inositol phosphate pools. Our long-term objective is to define how mood stabilizers work and to find common therapeutic targets in the pathophysiology of BD so new medications can be designed. Our specific aims are: 1) Determine the changes in abundance and activity of the newly-identified isoforms of IPS in brain regions after chronic lithium and valproate; 2) Determine the effects of lithium and valproate on IPS binding to clathrin and receptor-mediated endocytosis; 3) Establish the link between inositol metabolism and behavior in IPS-deficient mice; and 4) Identify drug-targetable convergent pathways for lithium valproate and lamotirgine responsive genes. To accomplish Aim 1 IPS and its isoforms will be quantified in drug-treated rat brain regions by enzyme activity assay Western blot and Q-PCR methods. To accomplish Aim 2 brain IPS and its complexes will be analyzed by Western and mass spectral analyses. Effects on receptor-mediated endocytosis will be monitored by following transferrin receptor trafficking and fluorescence resonance energy transfer (FRET) analysis. For Aim 3 lithium and valproate treated and untreated IPS knock-out mice will be subjected to standardized behavioral tests and their brains assayed for key enzymes in inositol metabolism. Aim 4 will be addressed by microarray and "pathway" analyses to map and compare the chronic lithium valproate and lamotrigine transcriptomes in human neuronal cell lines and in rat primary cortical neuronal cell cultures. Defining the mechanisms of lithium action on inositol metabolism in the brain from the perspective of IPS function may establish a direct connection between inositol regulation and behavior. Thus advancing our understanding of the pathophysiology of BD and disclosing new therapeutic targets for treating BD.

project_terms:
Address;Adverse effects;Affect;Behavior;Behavior Disorders;Behavioral;Binding (Molecular Function);Biochemical;Biological Assay;Biological Process;Bipolar Disorder;Brain;Brain region;Cell Culture Techniques;Cell Line;Cell physiology;Cells;Cerebellum;Chronic;Clathrin;Clinical;Complex;Corpus striatum structure;Data;Drug Targeting;Drug resistance;Endocytosis;Enzymatic Biochemistry;Enzymes;Equilibrium;Fluorescence Resonance Energy Transfer;Foundations;Functional disorder;Future;Gene Expression Profile;Generations;Genes;Global Change;Glucose;Goals;Health;Hippocampus (Brain);Homeostasis;Human;In Vitro;Individual;Inositol;Inositol Metabolism Pathway;Inositol Phosphates;Iraq;Knockout Mice;Link;Lithium;Manic;Manic Disorder;Maps;Measurement;Measures;Mediating;Mental Depression;Messenger RNA;Methods;Modeling;Monitor;Mood stabilizers;Moods;Mus;Mutation;Neurons;Pathway interactions;Patients;Pharmaceutical Preparations;Phospholipids;Phosphorylation;Predisposition;Prefrontal Cortex;Protein Isoforms;Proteins;Psychiatry;RNA;Rattus;Regulation;Relative (related person);Resistance;Route;Services;Signal Pathway;Signal Transduction;System;Testing;Therapeutic;Therapeutic Effect;Time;Transferrin Receptor;Treatment Efficacy;Veterans;War;Western Blotting;Work;abstracting;behavior test;brain pathway;chromatin remodeling;design;diagnostic accuracy;enzyme activity;feeding;gene interaction;improved;lamotrigine;membrane synthesis;mouse model;myo-Inositol-1-Phosphate Synthase;myo-inositol-1 (or 4)-monophosphatase;new therapeutic target;novel;protein protein interaction;public health relevance;receptor mediated endocytosis;research study;response;sugar;therapeutic target;trafficking;valproate

public_health_relevance:
PROJECT NARRATIVE (Potential Relevance to Veterans' Health) Bipolar Disorder (BD) is a major clinical problem for Veterans and there are many Iraq War Veterans treated by VA Psychiatry Service for lithium-resistant BD. To better manage these patients we must be able to identify common targets of mood-stabilizers so that future medications can be designed that have greater efficacy and fewer side effects. One way lithium can correct BD is by modulating brain inositol signal pathways. The brain maintains inositol (a sugar) homeostasis through three routes. Inositol phosphate synthase (IPS) is the only biosynthetic means of generating inositol from glucose. IPS is a new target linked to the genesis of BD. Stimulators of brain IPS enzyme which will then cause the generation of inositol inside the human brain can be considered as an important new class of mood stabilizing medications. The results from these studies will provide a highly positive impact on the health of Veterans with BD.


application_id: 8394590
project_number: 5I01BX000216-04
project_title: Nicotine-induced fibronectin expression in lung injury and repair
administering_ic: VA
award_notice_date: 2013-07-03
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000216
project_start_date: 2009-10-01
project_end_date: 2014-03-31
study_section: Respiration[PULM]
contact_pi_person_id: 1876531
contact_pi: ROMAN, JESSE
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2013-04-01
project_abstract:
DESCRIPTION (provided by applicant):  ABSTRACT: Nicotinic acetylcholine receptors (nAChRs) are pentameric channels expressed by neuronal and non-neuronal cells that are capable of signal transduction. Although much of what is known about the function of nAChRs relates to the nervous system information about their roles in non-neuronal systems is rapidly emerging. nAChRs are expressed in lung and interesting work performed in non-human primates suggests that nAChR activation by nicotine leads to pulmonary structural/functional abnormalities. We explored the mechanisms by which nAChR activation leads to matrix expression and found that this effect is mediated through a specific subgroup of nAChRs termed 17 nAChRs. Upon activation of these receptors in lung fibroblasts we observed induction of the MEK-1/Erk pathway and phosphorylation of the transcription factor CREB. In turn CREB stimulated the expression of fibronectin a matrix glycoprotein implicated in lung injury and repair. These events were blocked by 17 nAChR antagonists and were not present in 17 nAChR KO primary lung fibroblasts. In addition we found that nAChR activation stimulates lung fibroblast proliferation through MEK-1/Erk-dependent pathways. Similar observations were made in mice chronically stimulated with nicotine but not in developing and adult 17 nAChR KO animals. More recently we documented an increase in the proliferation and expression of fibronectin in fibroblasts exposed to cigarette smoke extract and observed an increase in fibronectin protein deposition in the lungs of mice exposed to tobacco smoke confirming the relevance of this process to tobacco-related disease. We also found that lung repair mechanisms appeared to be impaired after bleomycin-induced lung injury in the setting of chronic exposure to nicotine. These data unveil a novel role for non-neuronal nAChRs in control of lung tissue remodeling. Based on the above we hypothesize that 17 nAChR activation in lung promotes tissue remodeling by stimulating the proliferation of fibroblasts and their overexpression of matrix components like fibronectin through MEK-1/Erk-dependent signals. At baseline this leads to subtle alterations in matrix composition without causing overt consequences such as fibrosis. In contrast in the setting of lung injury this leads to exaggerated tissue remodeling that promotes inefficient repair and fibrosis. This will be tested in specific aims designed to: 1) Examine the role of 17 nAChRs in the proliferation myofibroblast transdifferentiation and matrix expression of lung fibroblasts and the role of MEK-1/Erk in mediating these events in wildtype vs. 17 nAChR KO lung fibroblasts 2) Examine the role of 17 nAChRs in lung injury and repair using wildtype and 17 nAChR KO mice in two models of lung injury (bleomycin model and the model of heterologous tracheal transplantation) and 3) Examine the effect of persistent 17 nAChR activation in lung repair after injury.

project_terms:
Adult;Affect;Animals;Bleomycin;Bronchoalveolar Lavage Fluid;Bronchopulmonary Dysplasia;CREB1 gene;Cells;Childhood;Chronic;Chronic Obstructive Airway Disease;Clinical Research;Collagen;Connective Tissue;Data;Deposition;Disease;Event;Exposure to;Extracellular Matrix;Fibroblasts;Fibronectins;Fibrosis;Genes;Genetic Transcription;Glycoproteins;Hamman-Rich syndrome;Harvest;Health;In Vitro;Inbred C57BL Mice;Injury;Ion Channel;Knock-out;Knockout Mice;Link;Lung;Lung diseases;Mediating;Messenger RNA;Modeling;Morphogenesis;Mus;Mutation;Myofibroblast;Neuroglia;Neurons;Nicotine;Nicotinic Receptors;Pathway interactions;Phosphorylation;Play;Population;Process;Production;Promotor (Genetics);Protein Isoforms;Proteins;Receptor Activation;Relative (related person);Reporting;Role;Signal Transduction;Site;Structure of parenchyma of lung;Subgroup;System;Testing;Time;Tissues;Tobacco;Tobacco smoke;Tobacco use;Transplantation;Veterans;Work;alpha-bungarotoxin receptor;base;cigarette smoke;design;drinking water;feeding;human MAP2K1 protein;in vivo;injured;injury and repair;interest;lung development;lung injury;lung repair;mTOR Signaling Pathway;nonhuman primate;novel;outcome forecast;overexpression;prenatal;protein expression;public health relevance;receptor;relating to nervous system;repaired;response;transcription factor;transdifferentiation


application_id: 8391631
project_number: 5I01BX001014-03
project_title: Regulation of adipose cells by autotaxin / lysophosphatidic acid signaling
administering_ic: VA
award_notice_date: 2012-12-12
opportunity_number: BX-10-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 001014
project_start_date: 2010-10-01
project_end_date: 2014-09-30
study_section: Endocriniology A[ENDA]
contact_pi_person_id: 1932626
contact_pi: SMYTH, SUSAN S.
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2012-10-01
project_abstract:
DESCRIPTION (provided by applicant):     Project Summary/Abstract Obesity and obesity-related cardiovascular complications are an increasing health burden in the United States. An analysis of nearly 2 million veterans receiving care at VA Medical Centers in 2000 indicated that ~68% were overweight and ~37% were obese. Despite extensive investigation viable therapeutic strategies to prevent weight gain or promote weight loss remain largely elusive. Attention has recently focused on the exciting possibility that a therapeutic strategy aimed at promoting energy expenditure by increasing brown adipose tissue (BAT) content or activity in adults could reduce fat deposition and obesity. However such a therapeutic strategy requires a comprehensive understanding of factors that regulate BAT. Lysophosphatidic acid (LPA) is a bioactive lipid present in physiologically relevant levels in plasma and other biologic fluids and is positioned to play a role in human health and disease. The secreted lysophospholipase D autotaxin (autotaxin/lysoLPD) is responsible for production of extracellular biologically active LPA which is in turn inactivated by enxymatic dephosphorylation by lipid phosphate phosphatases (LPP). We provide evidence that treatment of primary cultures of preadipocytes with LPA or autotaxin reduces expression of markers of brown adipocyte differentiation and LPP3 an endogenous regulator of Wnt; whereas a novel autotaxin/lysoPLD inhibitor dramatically promotes brown adipocyte differentiation. In mice overexpression of autotaxin reduces UCP1 expression and promotes diet-induced obesity. We therefore propose the central hypothesis that signaling pathways regulated by autotaxin and LPP3 contribute to diet-induced thermogenesis and regulate the development of diet-induced obesity. We will test our central hypothesis using state-of-the art genetic and pharmacologic approaches in three specific aims. In Aim One we will identify the role of autotaxin/lysoPLD in regulation of brown adipose tissue and diet-induced obesity in animal models. In Aim Two we will establish the molecular mechanism(s) by which LPA and autotaxin exert their effects. In Aim Three we identify the role of LPP3 in development and function of adipose tissue and diet-induced obesity. The aims of this grant provide a vehicle to address a major unresolved issue in the field of lysolipid signaling namely role of autotaxin/lysoPLD LPA and LPP3 in regulation of adiposity. These results will be significant because they are expected to provide innovative targets and provide proof-of-concept for novel inhibitors that may be used for prevention and treatment of obesity in humans.

project_terms:
Address;Adipocytes;Adipose tissue;Adult;Affect;American;Animal Model;Attention;Attenuated;Behavior Therapy;Body Weight decreased;Body mass index;Brown Fat;Cardiovascular system;Caring;Cell surface;Consumption;Data;Deposition;Desire for food;Development;Diet;Disease;Down-Regulation;Energy Metabolism;Exhibits;Family;Fatty acid glycerol esters;G-Protein-Coupled Receptors;Gap Junctions;Generations;Genetic;Goals;Grant;Health;Healthcare;Heating;Human;In Vitro;Integral Membrane Protein;Investigation;Knowledge;Lipids;Liquid substance;Lysophospholipids;Medical center;Molecular;Mus;Names;Obesity;Organism;Overweight;Plasma;Play;Positioning Attribute;Prevention;Production;Protein Dephosphorylation;Reagent;Recombinants;Regulation;Reporting;Research;Role;Signal Pathway;Signal Transduction;System;Techniques;Testing;Therapeutic;Thermogenesis;Tissue Differentiation;United States;Veterans;Weight Gain;Work;abstracting;adipocyte biology;adipocyte differentiation;alkylglycerophosphoethanolamine phosphodiesterase;extracellular;functional status;improved;in vivo;inhibitor/antagonist;innovation;lipid phosphate phosphatase;lysophosphatidic acid;member;novel;obesity prevention;obesity treatment;overexpression;precursor cell;prevent;public health relevance;tool;transdifferentiation


application_id: 8391581
project_number: 5I01BX000693-03
project_title: Airway sensors with myelinated afferents
administering_ic: VA
award_notice_date: 2012-12-12
opportunity_number: BX-10-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000693
project_start_date: 2010-10-01
project_end_date: 2014-09-30
study_section: Respiration[PULM]
contact_pi_person_id: 2084836
contact_pi: YU, JERRY (JUN)
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2012-10-01
project_abstract:
DESCRIPTION (provided by applicant):     The long-term objective of this proposal is to identify the unknown morphology and pathophysiological functions of pulmonary sensory receptors potentially benefiting more than 30 million patients with chronic obstructive pulmonary disease (COPD) and other pulmonary diseases in the US including veterans. Dyspnea bronchial inflammation hyperreactivity coughing and mucus secretion occur when airway sensory receptors are activated. These are common clinical manifestations of pulmonary disorders; however the underlying neural mechanisms remain unclear in part because our knowledge of airway receptor morphology and sensory behavior is limited. Increasing evidence points to important roles of myelinated airway receptors in pulmonary diseases. In this application we will test the hypothesis that vagal airway sensory units are not only transducers but also processors. Sensory units' behaviors depend on their structures. In addition a single unit may contain homogeneous or heterogeneous types of receptors providing varied mixed behavior. In this application we will address three specific aims to characterize the morphology and sensory behavior of myelinated airway receptors. 1) To identify different morphological structures of myelinated airway receptors by examing receptor morphology by immunohistochemical or neural tracing techniques combined with confocal microscopy. 2) To assign a specific receptor structure to physiologically defined classes of receptors by first electrophysiologically characterizing each known receptor type then tissue in the receptive field will be isolated dissected and immunohistochemically stained for receptor morphology. 3) To explore mechanosensor structure-function relationships and sensory integration mechansims by examining sensory behavior before and after blocking or stimulating one of the receptors in the unit. Thus integration mechanisms of sensory units can be explored. Successful completion of these studies will provide important structure-function relationships and establish a comprehensive objective classification system for the airway receptors. The presence of heterogenous receptors challenges the current view of the airway sensory unit. Transmission of heterogeneous information in a single unit verifies that the unit is not merely a transducer but also a processor. Our data will lay the foundation for future research and facilitate our understanding of the receptor function and their roles in pulmonary diseases and may help improve managements for cardiopulmonary patients suffering from cardiac failure COPD asthma acute lung injury and other pulmonary diseases.

project_terms:
Address;Adult Respiratory Distress Syndrome;Asthma;Behavior;Bronchoconstriction;C Fiber;Cardiopulmonary;Chemicals;Chronic Obstructive Airway Disease;Classification;Clinical;Confocal Microscopy;Cor pulmonale;Coughing;Data;Development;Disease;Dyspnea;Exhibits;Fiber;Foundations;Heart Diseases;Heart failure;Inflammation;Knowledge;Laboratories;Lung;Lung Injury Acute;Lung diseases;Mechanics;Mechanoreceptors;Microscopy;Morphology;Mucous body substance;Neurons;Patients;Physiological;Play;Process;Research;Role;Sensory;Sensory Receptors;Staining method;Stains;Structure;Structure of parenchyma of lung;Structure-Activity Relationship;Surveys;System;Techniques;Testing;Tissues;Trachea;Transducers;Vagus nerve structure;Veterans;improved;neuromechanism;operation;public health relevance;pulmonary function;receptive field;receptor;receptor function;relating to nervous system;response;sensor;sensory integration;sensory mechanism;transmission process

public_health_relevance:
Airway sensory receptors play important roles in cardiopulmonary diseases. Understanding their mechanisms of operation may help patient management thus benefitting a large number of veterans who suffer from cardiac and lung diseases including cardiac failure acute respiratory distress syndrome (ARDS) chronic obstructive pulmonary disease (COPD) and asthma.


application_id: 8391579
project_number: 5I01BX000672-04
project_title: Class B Scavenger Receptors and Atherosclerosis
administering_ic: VA
award_notice_date: 2012-12-12
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000672
project_start_date: 2009-10-01
project_end_date: 2013-09-30
study_section: Cardiovascular Studies A[CARA]
contact_pi_person_id: 7875298
contact_pi: VAN DER WESTHUYZEN, DENEYS REM
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2012-10-01
project_abstract:
DESCRIPTION (provided by applicant):   
  
The roles of the Class B scavenger receptors SR-BI and CD36 in the development of atherosclerotic disease a major cause of death remain poorly understood. This is due in part to the varied and still little understood functions of these two pattern recognition receptors in host defense and tissue homeostasis. The role of CD36 in the initiation of macrophage foam cell formation is controversial although CD36 has the ability to take up modified lipoproteins such as oxidized LDL. Other potentially important functions of CD36 include its role in the innate immune system and its role in inflammatory signaling. Atherosclerosis is a chronic inflammatory disease and we propose that CD36 plays a more complex role in atherosclerotic lesion formation. The structurally closely related receptor SR-BI plays a key role in reverse cholesterol transport to the liver and has been shown in several mouse models to be anti-atherogenic. Recent findings have also indicated additional extrahepatic roles of SR-BI in atherogenesis including effects in macrophages. The functional basis for these effects is not known although we have shown that SR-BI-null mice have a markedly enhanced inflammatory cytokine response and higher lethality in response to lipopolysaccharide (LPS). Furthermore mice deficient in both SR-BI and CD36 unexpectedly exhibit much greater atherosclerotic lesion formation than mice lacking either SR-BI or CD36 only. These findings indicate that SR-BI and CD36 each have athero-protective functions yet to be defined. The protective functions of the two receptors may be overlapping allowing at least partial compensation in mice lacking only one of the two receptors. The central hypothesis of this proposal is that SR-BI and CD36 act in a complementary or synergistic manner to protect against atherosclerosis. We propose that SR-BI and CD36 are key to the innate defense against atherosclerosis a chronic inflammatory disease. The following 4 objectives are proposed to examine these hypotheses: Objective 1 will examine the complementary roles SR-BI and CD36 in atherosclerotic lesion formation in response to high fat diets. This will be accomplished by analyzing the extent and nature of aortic lesions in wild type SR-BI-null CD36-null and SR-BI-nullXCD36-null mice. Objective 2 will investigate the roles of macrophage-expressed Class B scavenger receptors in atherosclerosis. This will be accomplished through the use of bone marrow transplantation. Objective 3 will identify the complementary or synergistic roles of SR-B1 and CD36 in macrophage foam cell formation. This will be accomplished by measuring lipid accumulation in wild type and SR-BI-XCD36- DKO macrophages treated with modified LDLs. Cells will be examined for lipid related protein expression and by gene microarray analysis. Lipid uptake and efflux pathways will be compared. Objective 4 will investigate enhanced inflammatory signaling in CD36/SR-BI DKO macrophages and its effects on macrophage lipid accumulation. The impact of inflammatory signaling on modified LDL uptake in primary macrophages will be evaluated together with the hypothesis that increased plasma membrane cholesterol content in SR-BIxCD36 DKO macrophages stimulates inflammatory signaling pathways. These studies are expected to provide new understanding of the metabolic functions of CD36 and SR-BI as well as their role in atherosclerosis.

project_terms:
Affect;Antiatherogenic;Arterial Fatty Streak;Arteries;Atherosclerosis;Bone Marrow Transplantation;CD36 gene;Cardiovascular Diseases;Cause of Death;Cell membrane;Cells;Cholates;Cholesterol;Chronic;Complex;Development;Diet;Disease;Exhibits;Extrahepatic;Fatty acid glycerol esters;Financial compensation;Foam Cells;Genes;Hepatic;Homeostasis;Host Defense;Immune system;Inflammation;Inflammatory;Inflammatory Response Pathway;Knock-out;Knockout Mice;LDL Cholesterol Lipoproteins;Lesion;Lipids;Lipopolysaccharides;Lipoprotein Receptor;Lipoproteins;Liver;Low-Density Lipoproteins;Measures;Metabolic;Metabolism;Microarray Analysis;Mus;Nature;Pathway interactions;Pattern recognition receptor;Plasma;Play;Regulation;Risk Factors;Role;SR-B proteins;SR-BI receptor;Signal Pathway;Signal Transduction;TLR4 gene;Testing;Tissues;Transplantation;United States;Veterans;atherogenesis;base;drug development;feeding;macrophage;mouse model;oxidized low density lipoprotein;protein expression;public health relevance;receptor;research study;response;reverse cholesterol transport;scavenger receptor;treatment strategy;uptake

public_health_relevance:
Cardiovascular disease remains the number one killer in the United States and among veterans. Although widely studied the mechanisms underlying atherosclerosis are not yet clearly understood. Plasma cholesterol and LDL levels are a major risk factor for atherosclerosis but the underlying mechanisms for this are still poorly understood. Further understanding of how LDL cholesterol accumulates in the artery wall and the roles of the various receptors for lipoproteins will potentially allow for the identification of new treatment strategies and new drug development.


application_id: 8391566
project_number: 5I01BX000622-04
project_title: Mechanisms of Renal Lipid Accumulation in Diabetic Nephropathy
administering_ic: VA
award_notice_date: 2012-12-12
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000622
project_start_date: 2009-10-01
project_end_date: 2013-09-30
study_section: Nephrology[NEPH]
contact_pi_person_id: 3123430
contact_pi: TANNOCK, LISA R
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2012-10-01
project_abstract:
DESCRIPTION (provided by applicant):     Nephropathy is a major complication of both type 1 and type 2 diabetes which causes considerable morbidity and mortality. Despite best current treatments significant numbers of individuals with albuminuria progress to end stage renal disease. Although the mechanisms underlying this progression are not fully understood renal lipid and lipoprotein accumulation has been shown to accelerate the development of nephropathy. Renal lipid accumulation triggers an influx of inflammatory cells with subsequent development of glomerulosclerosis the characteristic lesion of diabetic nephropathy. Glomerulosclerosis is characterized by increased deposition of mesangial matrix including proteoglycans. Of particular interest the renal content of the small leucine-rich proteoglycan (SLRP) biglycan is increased in diabetes. TGF-2 which is elevated in diabetes and is known to be a key mediator of diabetic nephropathy development and progression also increases mesangial matrix deposition including increased expression of biglycan. Furthermore we have shown that TGF-2 increases the size and LDL binding affinity of renal proteoglycans. Thus increased renal proteoglycan (biglycan) synthesis induced by elevated TGF-2 in diabetes may be responsible for mediating the renal accumulation of lipoproteins. The overall goal of this grant is to test the hypothesis that renal lipid accumulation is mediated through interactions of lipoproteins with renal proteoglycans especially biglycan. This will be tested by comparing diabetic nephropathy in mice expressing proteoglycan-binding defective LDL with littermates expressing wildtype LDL. To determine if biglycan is the key proteoglycan responsible diabetic nephropathy will be compared between biglycan deficient and wildtype mice. The experiments outlined in this grant will provide direct in vivo experimental data identifying if proteoglycan mediated renal lipid accumulation contributes significantly to the development and progression of diabetic nephropathy and if the key proteoglycan is biglycan. This grant will also identify if biglycan serves as a natural inhibitor of TGF-2 in vivo. Thus the significance of this proposal is that it not only identifies a mechanism linking hyperlipidemia and diabetic nephropathy but also will identify novel targets to prevent or intervene in the development of diabetic nephropathy.

project_terms:
Affinity;Albuminuria;Animal Model;Antibodies;Binding (Molecular Function);Biological Models;Cells;Characteristics;Clinical Research;Complication;Complications of Diabetes Mellitus;Data;Deposition;Development;Diabetes Mellitus;Diabetic Nephropathy;Enalapril;End stage renal failure;Goals;Grant;Healthcare;High Prevalence;Hyperlipidemia;In Vitro;Individual;Inflammatory;Kidney;Kidney Diseases;Lesion;Leucine;Link;Lipids;Lipoproteins;Low Density Lipoprotein Receptor;Low-Density Lipoproteins;Mediating;Mediator of activation protein;Modeling;Morbidity - disease rate;Mortality Vital Statistics;Mus;Proteoglycan;Reporting;Research;Role;Testing;Therapeutic Intervention;Tissues;Transforming Growth Factor beta;Translating;Up-Regulation (Physiology);Veterans;biglycan;diabetic;glomerulosclerosis;in vivo;inhibitor/antagonist;interest;mouse model;nephrogenesis;neutralizing antibody;novel;prevent;public health relevance;receptor binding;research study;type I and type II diabetes

public_health_relevance:
PUBLIC HEALTH RELEVANCE:     Potential Impact on Veterans Health Care: Veterans have very high prevalence of diabetes hyperlipidemia and diabetes complications including nephropathy. This research will either identify or exclude proteoglycans as a cause of renal lipid accumulation contributing to diabetic nephropathy. The grant will prove the concept in an animal model system and the results can be easily translated into clinical research. The ultimate goal is to identify novel targets for therapeutic intervention to decrease the morbidity and mortality of diabetic nephropathy.


application_id: 8391562
project_number: 5I01BX000610-04
project_title: Analysis of PTH and Dopamine Receptor Signaling in Proximal Tubules
administering_ic: VA
award_notice_date: 2012-12-12
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000610
project_start_date: 2009-10-01
project_end_date: 2013-09-30
study_section: Nephrology[NEPH]
contact_pi_person_id: 2262379
contact_pi: LEDERER, ELEANOR D
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2012-10-01
project_abstract:
DESCRIPTION (provided by applicant):  Project Summary/Abstract The goal of this project is to determine how the structural protein NHERF-1 sodium-hydrogen exchanger regulatory factor isoform 1 regulates the trafficking of Npt2a the type IIa sodium phosphate cotransporter to the apical membrane of the renal proximal tubule. The expression of Npt2a at the apical membrane is a critical regulatory step because the level of expression and function of Npt2a is the primary regulator of total body phosphate homeostasis. VA-funded research from this laboratory had revealed that the absence of NHERF-1 in a model of proximal renal tubule OK (opossum kidney) cells resulted in a marked decrease in Npt2a expression. The decrease in expression was due to two factors 1) decreased transcription of Npt2a and 2) decreased trafficking of Npt2a to the apical membrane. The mechanisms for the faulty trafficking of Npt2a have not been determined and are the subject of this proposal. Npt2a translated in the NHERF-deficient cells (OK-H) lacked a critical post-translational modification glycosylation. These proteins instead of trafficking to apical membrane accumulated in a perinuclear location. Inefficient apical membrane localization had previously been described in a mouse lacking expression of NHERF-1. Expression of a NHERF-1 construct that lacked a normal PDZ-2 domain did not allow trafficking of Npt2a to the apical membrane. NHERF-1 is a multiple PDZ domain protein which interacts with multiple signaling molecules and transporter proteins. A role for the PDZ-2 domain in Npt2a regulation had not been observed previously. Finally inhibition of SNARE (SNAP Receptor) protein interaction using a competing peptide introduced into OK cells blocked insertion of Npt2a into the apical membrane demonstrating that Npt2a is transported to the apical membrane via a vesicular transport mechanism. The preliminary data suggested the hypothesis that NHERF-1 is essential for the forward trafficking of Npt2a from site of synthesis to the apical membrane. First the role of the post-translational modifications glycosylation and phosphorylation on Npt2a apical membrane trafficking will be examined by determining the cellular localization of transfected Npt2a constructs lacking the motifs required for these post translational modifications in a cell culture model. The glycosylation phosphorylation and ubiquitination states of Npt2a will be compared in NHERF-replete and NHERF-deficient OK cells and in wild type and NHERF-1 knock out mouse proximal tubule cells. The effect of inhibitors of glycosylation on the intracellular localization of Npt2a will be determined. Second the intracellular site or sites of interaction between NHERF-1 and Npt2a will be analyzed. Forward trafficking of Npt2a labeled by a GFP tag and by S35 methionine in OK cells and OK-H cells will be slowed by culture at 20 C. Analysis of Npt2a localization by confocal imaging density centrifugation and immunoelectronmicroscopy will be performed at sequential time points until the proteins are detected as biotinylated forms indicating appropriate insertion into the apical membrane. Trafficking of Npt2a in the two cell culture models will be compared. Proteins associated with Npt2a at each time point will be determined by immunoprecipitation and proteomic analysis. To define which steps in the forward trafficking are NHERF- dependent Npt2a trafficking will be compared in NHERF-replete and NHERF-deficient cells under conditions of traffic arrest: ezrin deficiency inhibition of SNARE interaction and CLC-5 (intracellular chloride channel CLC family isoform 5) deficiency. Third the sites on Npt2a and NHERF-1 critical for the NHERF-1 effect on Npt2a trafficking will be determined by mutational analysis of both proteins followed by both in vitro analysis of protein interaction and assessment of intracellular interaction by FRET methodology. These studies will define where NHERF-1 acts in Npt2a forward trafficking define the sites on both proteins responsible for their interaction and yield mechanistic insights for this unique functional process.

project_terms:
Address;Affect;Age;Apical;CLC Gene;Cardiovascular Diseases;Carrier Proteins;Cell Culture Techniques;Cell Line;Cells;Centrifugation;Chloride Channels;Country;Data;Didelphidae;Disease;Dopamine Receptor;Event;Exhibits;Family;Fluorescence Resonance Energy Transfer;Funding;Genetic Transcription;Goals;Health;Hip Fractures;Homeostasis;Image;Immunoprecipitation;In Vitro;Individual;Kidney;Kidney Calculi;Knockout Mice;Label;Laboratories;Laboratory Research;Location;Medical;Membrane Microdomains;Membrane Protein Traffic;Membrane Transport Proteins;Metabolic Bone Diseases;Methionine;Methodology;Modeling;Molecular;Morbidity - disease rate;Mortality Vital Statistics;Mus;Mutation;Pathway interactions;Peptides;Phosphorylation;Physiologic pulse;Play;Population;Post-Translational Protein Processing;Process;Protein Analysis;Protein Isoforms;Proteins;Proteomics;Proximal Kidney Tubules;Receptor Signaling;Regulation;Renal function;Research;Research Personnel;Role;SNAP receptor;Signaling Molecule;Site;Staging;Structural Protein;Tertiary Protein Structure;Time;TimeLine;Transfection;Translating;Transport Vesicles;Tubular formation;Ubiquitination;Vesicle;Veterans;Work;abstracting;apical membrane;base;cardiovascular risk factor;caveolin 1;cold temperature;density;design;ezrin;glycosylation;in vitro Assay;inhibitor/antagonist;inorganic phosphate;insight;kidney cell;mutant;protein expression;public health relevance;receptor;research study;sodium-hydrogen exchanger regulatory factor;sodium-phosphate cotransporter proteins;solute;syntaxin binding protein 1;trafficking;wasting

public_health_relevance:
PUBLIC HEALTH RELEVANCE:  Project Narrative The goal of this project is to determine how the protein NHERF-1 sodium-hydrogen exchanger regulatory factor isoform 1 regulates the trafficking of Npt2a the type IIa sodium phosphate cotransporter to the apical membrane of the renal proximal tubule. The kidney regulates the total body content of phosphate through altering the expression of Npt2a at the apical membrane. Phosphate wasting can cause kidney stones and metabolic bone disease while phosphate retention is a risk factor for cardiovascular events common diseases in Veterans of all ages. Cardiovascular disease is the major cause of morbidity and mortality in the Veteran population. Kidney stones tend to affect individuals during their most productive years resulting in over $2 billion dollars of medical expenses in this country. Metabolic bone disease contributes substantially to the occurrence of hip fracture a disabling event with up to a 50% one year mortality. Thus understanding how the kidney regulates phosphate homeostasis has significant relevance to the health of Veterans of all ages.


application_id: 8391138
project_number: 5I01BX000350-04
project_title: Alcohol Proteasome Dysfunction in Liver Injury
administering_ic: VA
award_notice_date: 2012-12-11
opportunity_number: BX-08-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000350
project_start_date: 2009-04-01
project_end_date: 2013-09-30
study_section: Gastroenterology[GAST]
contact_pi_person_id: 1902075
contact_pi: MCCLAIN, CRAIG J.
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2012-10-01
project_abstract:
DESCRIPTION (provided by applicant):     Studies from our VA-supported research and that of others demonstrate a critical role for the proinflammatory cytokine tumor necrosis factor (TNF) in the development and progression of alcoholic liver disease (ALD). Alcoholic liver disease is associated with increased oxidative stress and impaired hepatocyte 26S proteasome function. Our preliminary data demonstrate that inhibition of 26S proteasome function can sensitize hepatocytes to TNF cytotoxicity and higher doses of proteasome inhibitors directly induce hepatocyte death. Additionally this apoptotic death involves the induction of lysosomal membrane permeability (LMP) and is mediated by lysosomal proteases (cathepsins B and D). Based on the increasingly evident role of cathepsins as key downstream mediators of apoptosis and our preliminary results we postulate that chronic alcohol abuse with subsequent proteasome dysfunction leads to LMP and release of lysosomal cathepsins which induce hepatocyte cell death and further sensitize hepatocytes to TNF-mediated hepatotoxicity. Neutrophil infiltration is a hallmark of alcoholic hepatitis and infiltrating neutrophils are thought to play an etiologic role in the liver injury in ALD. We recently demonstrated that inhibition of proteasome function in hepatocytes induces and markedly enhances the TNF-inducible expression of the pro-inflammatory and neutrophil chemotactic cytokine Interleukin-8 (IL-8). It is our working hypothesis that chronic alcohol abuse mediated proteasome dysfunction is associated with increased hepatocyte IL-8 production which significantly contributes to neutrophil infiltration and hepatic injury in ALD. The overall objective of this proposal is to elucidate the cellular and molecular mechanism(s) involved in proteasome dysfunction-mediated hepatic injury associated with ALD. The specific objectives of this proposal are to: 1) Determine the effects of proteasome inhibition and ethanol on sensitization to TNF hepatotoxicity 2) Determine the role of lysosomal membrane permeability and cathepsins B and D in the proteasome dysfunction-mediated hepatocyte apoptosis and sensitization to TNF hepatotoxicity and 3) Determine the effects of proteasome inhibition and ethanol on the hepatocyte expression of the critical neutrophil chemotactic CXC cytokine IL-8. Potential Impact on Veterans Health: Alcoholic liver disease is a leading cause of death from liver disease in the United States and in the Veterans Administration. In studies from the Veterans Administration patients with cirrhosis and superimposed alcoholic hepatitis had > 60% mortality over a 4 year period of time with most of those deaths occurring in the first month. Thus the prognosis for this disease is more ominous than for many common types of cancer such as breast prostate and colon. The longterm goals of this laboratory are to define mechanism(s) for liver cell death in ALD that will provide new insights into this process and provide new areas for therapeutic intervention> This is especially important in ALD where there is no FDA-approved therapy. The potential clinical impact of this work is high.

project_terms:
26S proteasome;Affect;Alcohol abuse;Alcoholic Hepatitis;Alcoholic Liver Cirrhosis;Alcoholic Liver Diseases;Alcoholic beverage heavy drinker;Alcohols;Animals;Antioxidants;Apoptosis;Apoptotic;Area;Breast;Calpain;Cathepsins;Cathepsins B;Cause of Death;Cell Death;Cell Membrane Permeability;Cells;Cessation of life;Chronic;Cirrhosis;Clinical;Colon;Data;Development;Disease;Dose;Ethanol;Ethanol Metabolism;FDA approved;Functional disorder;Future;Genetic;Goals;Grant;Health;Hepatic;Hepatocyte;Hepatotoxicity;In Vitro;Inflammatory;Interleukin-8;Label;Laboratories;Liver diseases;Lysosomes;Mediating;Mediator of activation protein;Mitochondria;Modeling;Molecular;Morbidity - disease rate;Mortality Vital Statistics;Mus;Neutrophil Infiltration;Oxidative Stress;Pathogenesis;Pathway interactions;Patients;Peptide Hydrolases;Permeability;Plasma;Play;Polyubiquitin;Process;Production;Prostate;Proteasome Inhibition;Proteasome Inhibitor;Proteins;Proteolysis;Reactive Oxygen Species;Reduced Glutathione;Research;Research Support;Resistance;Role;Route;Signal Transduction;Stimulus;Therapeutic Intervention;Time;Tumor Necrosis Factor-alpha;Ubiquitin;United States;United States Department of Veterans Affairs;Veterans;Vitamin E;Work;alcohol effect;base;cancer type;chemokine;chronic alcohol ingestion;clinically relevant;cytokine;cytotoxicity;insight;liver injury;monocyte;multicatalytic endopeptidase complex;neutrophil;outcome forecast;protein degradation;public health relevance;response

public_health_relevance:
PUBLIC HEALTH RELEVANCE:      The overall objective of this proposal is to elucidate the cellular and molecular mechanism(s) involved in alcohol and proteasome dysfunction-mediated hepatic injury associated with alcoholic liver disease (ALD). We expect that the results of our study will provide critical molecular insights into the pathogenesis of ALD and provide a basis for clinically relevant paradigms for future development of potential therapeutic intervention(s) in patients with ALD.


application_id: 8390433
project_number: 5I01CX000240-03
project_title: Mechanisms of hepcidin effects in the anemia of chronic disease
administering_ic: VA
award_notice_date: 2012-11-27
opportunity_number: CX-10-001
application_type: 5
activity_code: I01
ic: CX
serial_number: 000240
project_start_date: 2010-10-01
project_end_date: 2014-09-30
study_section: Hematology[HEMA]
contact_pi_person_id: 1876507
contact_pi: MEANS, ROBERT T
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2012-10-01
project_abstract:
DESCRIPTION (provided by applicant):     The anemia of chronic disease (ACD) is one of the most common clinical syndromes encountered in the practice of medicine. This disorder typically manifests itself as a hypoproliferative anemia accompanied by a low serum iron concentration despite adequate reticuloendothelial iron stores. ACD has generally been considered to result from a combination of pathologic processes which can be linked to the cytokine mediators of inflammation. However the diagnostic importance of altered iron metabolism in ACD has led many investigators to consider that this feature denotes the dominant pathophysiologic mechanism of this syndrome. Hepcidin is an antibacterial protein which is produced in the liver circulates in the blood and is excreted in the urine. It is a type II acute phase protein the expression of which is induced by interleukin (IL)-6 and down-regulated by tumor necrosis factor (TNF). The specific iron regulatory functions of hepcidin as well as the anemia syndromes observed in patients and transgenic animals with abnormalities of hepcidin gene expression (discussed below) have led many to consider that it is the key to unlocking "the mysteries of ACD ".  Three major processes are involved in the pathogenesis of ACD. A slight shortening of red cell survival creates a demand for a small increase in red cell production by the bone marrow. The marrow cannot respond adequately to this demand due to impaired erythropoiesis and impaired mobilization of reticuloendothelial system iron stores. A potential role for hepcidin in the iron anomalies of ACD is strongly supported by available data. However iron changes are not the sole abnormality characteristic of ACD. If hepcidin is the major factor responsible for ACD then it should also contribute to the impaired erythropoiesis observed in this syndrome.  The hypothesis that hepcidin contributes to impaired erythropoiesis in ACD will be addressed through the following specific aims: 1. Determination of hepcidin effects on erythroid colony formation and investigation of the mechanisms involved. This will include A. whether the effects of hepcidin on erythropoiesis are direct-acting or require an accessory cell; B. the role of Epo concentration in hepcidin effects on erythropoiesis; C. the role of apoptosis/anti-proliferation in inhibitory effects of hepcidin on erythropoiesis; D. the effects of cytokines on the hepcidin/erythropoiesis relationship; E. the contributions of iron to hepcidin effects on CFU-E colony formation; F. the role of ferroportin (FPN) and variant FPN in hepcidin effects on CFU-E; and G. effects of Epo and inflammatory cytokines on FPN variant expression in erythroid cells. 2. Determination of the interactions of hepcidin and cytokines in bone marrow macrophage iron transport. Hepcidin inhibits egress of iron from bone marrow macrophages by downregulation of the iron export protein FPN. The ability of inflammatory cytokines to enhance or diminish this effect will be determined as well as any effects on the expression of variant FPN.3. Determination of hepcidin effects on induced Epo production in vitro and delineation of involved mechanisms. This will include A. determination and quantification of the effects of hepcidin on the induction of Epo protein in hepatic and renal models and of the time-course of these effects; and B. determination of the roles of inflammatory cytokines in modulating these effects; C. the role of iron availability in these effects; and D. determination of the effects of hepcidin on Epo transcription and the mechanism involved in these effects. 4. Effects of rhEpo on hepcidin production in vitro. In this Aim we will determine A. the effects of Epo on hepcidin production by HepG2 as well as B. the extent to which these effects are modulated by TNF IL-1 and IL-6. Relevance to VA mission: The proposed studies can potentially lead to an enhanced understanding of the mechanisms of ACD and lead to more effective therapy.

project_terms:
Acute-Phase Proteins;Address;Anemia;Anemia due to Chronic Disorder;Anti-Bacterial Agents;Antigen-Presenting Cells;Apoptosis;Blood;Bone Marrow;CFU-E;Cell Survival;Characteristics;Chronically Ill;Clinical;Data;Diagnostic;Disease;Down-Regulation;Erythrocytes;Erythroid;Erythroid Cells;Erythropoiesis;Etiology;Gene Expression;Genetic Transcription;Hepatic;In Vitro;Inflammation Mediators;Inflammatory;Interleukin-1;Interleukin-6;Investigation;Iron;Kidney;Lead;Link;Liver;Marrow;Medicine;Mission;Modeling;Morbidity - disease rate;Mortality Vital Statistics;Pathogenesis;Pathologic Processes;Patients;Population;Process;Production;Protein Export Pathway;Proteins;Research Personnel;Reticuloendothelial System;Role;Serum;Syndrome;Time;Transgenic Animals;Tumor Necrosis Factor-alpha;Urine;Variant;cytokine;effective therapy;hepcidin;iron metabolism;macrophage;metal transporting protein 1;protein expression;public health relevance

public_health_relevance:
Anemia is a major contributor to morbidity and mortality in chronically ill patients and the anemia of chronic disease (ACD) is the major etiology of this anemia. This disorder typically manifests itself as a hypoproliferative anemia accompanied by a low serum iron concentration despite adequate reticuloendothelial iron stores. ACD is highly prevalent in the VA population. In ACD a slight shortening of red cell survival creates a demand for a small increase in red cell production by the bone marrow. The marrow cannot respond adequately to this demand due to impaired erythropoiesis and impaired mobilization of reticuloendothelial system iron stores. A potential role for hepcidin in the iron anomalies of ACD is strongly supported by available data. If hepcidin is the major factor responsible for ACD then it should also contribute to the impaired erythropoiesis observed in this syndrome. The proposed studies can potentially lead to an enhanced understanding of the mechanisms of ACD and therefore to more effective therapy.


application_id: 8259053
project_number: 5I01BX000387-03
project_title: Mood-Stabilizing Medications and the Inositol Signaling System
administering_ic: VA
award_notice_date: 2011-12-07
opportunity_number: BX-09-008
application_type: 5
activity_code: I01
ic: BX
serial_number: 000387
project_start_date: 2010-04-01
project_end_date: 2014-03-31
study_section: Neurobiology A[NURA]
contact_pi_person_id: 7118863
contact_pi: PARTHASARATHY, RANGA N
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2012-04-01
project_abstract:
DESCRIPTION (provided by applicant):     Project Summary/Abstract Bipolar disorder (BD) is a devastating condition affecting many Veterans. Lithium and valproate are the drugs of choice for the manic phase of BD. While therapeutically effective their mechanisms of action are not fully understood and undesirable side effects are common. Lithium inhibits brain inositol monophosphatase (IMPase) thereby depleting the available pool of free inositol and resulting in the correction of the manic phase of BD. Inositol synthase (IPS) is a key enzyme that regulates inositol levels by generating the precursor inositol phosphate (IP). Our preliminary data demonstrate that rat brain IPS is up-regulated by chronic lithium treatment which would have the same net effect as inositol depletion relative to inositol phosphate. We hypothesize that lithium and other mood stabilizers exert their therapeutic effects by up-regulating IPS expression increasing the available inositol phosphate pool. This implies that the pathophysiological mechanism underlying BD is a disturbance in maintaining a proper balance in the ratio of free inositol to inositol phosphate pools. Our long-term objective is to define how mood stabilizers work and to find common therapeutic targets in the pathophysiology of BD so new medications can be designed. Our specific aims are: 1) Determine the changes in abundance and activity of the newly-identified isoforms of IPS in brain regions after chronic lithium and valproate; 2) Determine the effects of lithium and valproate on IPS binding to clathrin and receptor-mediated endocytosis; 3) Establish the link between inositol metabolism and behavior in IPS-deficient mice; and 4) Identify drug-targetable convergent pathways for lithium valproate and lamotirgine responsive genes. To accomplish Aim 1 IPS and its isoforms will be quantified in drug-treated rat brain regions by enzyme activity assay Western blot and Q-PCR methods. To accomplish Aim 2 brain IPS and its complexes will be analyzed by Western and mass spectral analyses. Effects on receptor-mediated endocytosis will be monitored by following transferrin receptor trafficking and fluorescence resonance energy transfer (FRET) analysis. For Aim 3 lithium and valproate treated and untreated IPS knock-out mice will be subjected to standardized behavioral tests and their brains assayed for key enzymes in inositol metabolism. Aim 4 will be addressed by microarray and "pathway" analyses to map and compare the chronic lithium valproate and lamotrigine transcriptomes in human neuronal cell lines and in rat primary cortical neuronal cell cultures. Defining the mechanisms of lithium action on inositol metabolism in the brain from the perspective of IPS function may establish a direct connection between inositol regulation and behavior. Thus advancing our understanding of the pathophysiology of BD and disclosing new therapeutic targets for treating BD.    PUBLIC HEALTH RELEVANCE:     PROJECT NARRATIVE (Potential Relevance to Veterans' Health) Bipolar Disorder (BD) is a major clinical problem for Veterans and there are many Iraq War Veterans treated by VA Psychiatry Service for lithium-resistant BD. To better manage these patients we must be able to identify common targets of mood-stabilizers so that future medications can be designed that have greater efficacy and fewer side effects. One way lithium can correct BD is by modulating brain inositol signal pathways. The brain maintains inositol (a sugar) homeostasis through three routes. Inositol phosphate synthase (IPS) is the only biosynthetic means of generating inositol from glucose. IPS is a new target linked to the genesis of BD. Stimulators of brain IPS enzyme which will then cause the generation of inositol inside the human brain can be considered as an important new class of mood stabilizing medications. The results from these studies will provide a highly positive impact on the health of Veterans with BD.

project_terms:
Address;Adverse effects;Affect;Behavior;Behavior Disorders;Behavioral;Binding (Molecular Function);Biochemical;Biological Assay;Biological Process;Bipolar Disorder;Brain;Brain region;Cell Culture Techniques;Cell Line;Cell physiology;Cells;Cerebellum;Chronic;Clathrin;Clinical;Complex;Corpus striatum structure;Data;Drug Delivery Systems;Drug resistance;Endocytosis;Enzymatic Biochemistry;Enzymes;Equilibrium;Fluorescence Resonance Energy Transfer;Foundations;Functional disorder;Future;Gene Expression Profile;Generations;Genes;Global Change;Glucose;Goals;Health;Hippocampus (Brain);Homeostasis;Human;In Vitro;Individual;Inositol;Inositol Metabolism Pathway;Inositol Phosphates;Iraq;Knockout Mice;Link;Lithium;Manic;Manic Disorder;Maps;Measurement;Measures;Mediating;Mental Depression;Messenger RNA;Methods;Modeling;Monitor;Mood stabilizers;Moods;Mus;Mutation;Neurons;Pathway interactions;Patients;Pharmaceutical Preparations;Phospholipids;Phosphorylation;Predisposition;Prefrontal Cortex;Protein Isoforms;Proteins;Psychiatry;RNA;Rattus;Regulation;Relative (related person);Resistance;Route;Services;Signal Pathway;Signal Transduction;System;Testing;Therapeutic;Therapeutic Effect;Time;Transferrin Receptor;Treatment Efficacy;Veterans;War;Western Blotting;Work;abstracting;behavior test;brain pathway;chromatin remodeling;design;diagnostic accuracy;enzyme activity;feeding;gene interaction;improved;lamotrigine;membrane synthesis;mouse model;myo-Inositol-1-Phosphate Synthase;myo-inositol-1 (or 4)-monophosphatase;new therapeutic target;novel;protein protein interaction;receptor mediated endocytosis;research study;response;sugar;therapeutic target;trafficking;valproate

public_health_relevance:
PROJECT NARRATIVE (Potential Relevance to Veterans' Health) Bipolar Disorder (BD) is a major clinical problem for Veterans and there are many Iraq War Veterans treated by VA Psychiatry Service for lithium-resistant BD. To better manage these patients we must be able to identify common targets of mood-stabilizers so that future medications can be designed that have greater efficacy and fewer side effects. One way lithium can correct BD is by modulating brain inositol signal pathways. The brain maintains inositol (a sugar) homeostasis through three routes. Inositol phosphate synthase (IPS) is the only biosynthetic means of generating inositol from glucose. IPS is a new target linked to the genesis of BD. Stimulators of brain IPS enzyme which will then cause the generation of inositol inside the human brain can be considered as an important new class of mood stabilizing medications. The results from these studies will provide a highly positive impact on the health of Veterans with BD.


application_id: 8212755
project_number: 5I01BX000962-02
project_title: Exosomal TRAF2-CSN5 complex mediated inflammation promotes tumor growth
administering_ic: VA
award_notice_date: 2011-12-09
opportunity_number: BX-10-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000962
project_start_date: 2011-04-01
project_end_date: 2015-03-31
study_section: Immunology and Dermatology A[IMMA]
contact_pi_person_id: 1889248
contact_pi: ZHANG, HUANG-GE
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2012-04-01
project_abstract:
DESCRIPTION (provided by applicant):     The overall goal of this proposal is to elucidate the mechanisms by which lung tumor exosomes promote an inflammatory response namely the activation of macrophages and promote the progression and growth of lung tumors. Our preliminary data indicate that pretreatment of A/J mice with exosomes produced by TC-1 lung tumor cells or exosomes isolated from the lung tissue of A/J mice that have been pretreated with the carcinogen urethane results in more rapid tumor growth and earlier progression of lung carcinomas. Urethane treatment results in the recruitment of the activated form of TRAF2 to the exosomes and the exosomes with activated TRAF2 are taken up by the bone marrow-derived precursors of macrophages leading to their maturation and subsequent migration to the lung. The recruitment of the activated TRAF2 to the exosomes requires degradation of the inflammation suppressor protein CYLD in the tumor cells; moreover TRAF2 can mediate ubiquination of CYLD thereby promoting its degradation. The results of siRNA TRAF2 knockout indicated however that TRAF2 is required but is not sufficient for ubiquitination of CYLD. We have now identified two exosomal proteins that interact with TRAF2 Itch and CSN5 which have the potential to enhance the ubiquitination of CYLD in lung tumor cells. Our recent data indicating that CYLD is not packed in exosomes isolated from the peripheral blood of lung cancer patients although it is present in exosomes from healthy volunteers indicate the potential clinical relevance of this model that directly links the effects of a carcinogen with an inflammatory response and promotion of tumor development. We propose to: (1) Confirm the role of lung tumor exosomal TRAF2 in the promotion of urethane-induced lung carcinomas by determining whether knockout of TRAF2 in TC-1 lung tumor cells is sufficient for (i) inhibition of macrophage differentiation and (ii) prevention of lung tumor exosome-mediated enhancement of urethane induced lung cancer. (2) Determine if other exosomal proteins that interact with CYLD participate in the ubiquitination of CYLD in TC-1 tumor cells and determine if the elimination of these proteins stabilizes exosomal CYLD and suppresses exosomal TRAF2 activation resulting in the inhibition of macrophage activation and prevention of tumor growth. (3) Determine if macrophages pulsed with lung tumor exosomes isolated from lung cancer patients promote lung tumor growth in a NOD-SCID mouse model. The data generated should permit the development of a highly innovative model of the cellular and molecular basis for exosome-mediated chronic inflammation and promotion of tumor growth and indicate novel preventive and therapeutic strategies.    PUBLIC HEALTH RELEVANCE:     The proposed research is highly relevant to characterizing the association of lung tumor exosomes mediated inflammatory responses and lung tumor growth in that it is focused on characterizing the molecular mechanisms underlying the ability of tumor exosomes to both activate macrophages and promote tumor growth. The central hypothesis is that exosomes produced by lung tumor cells or lung tumor tissue are taken up by bone marrow myeloid precursor cells. These cells further differentiate into macrophages and migrate into the lung promote the progression of urethane- induced lung cancer.

project_terms:
A/J Mouse;Acceleration;Anti-Inflammatory Agents;Bone Marrow;Bone Marrow Cells;Cancer Patient;Carcinogens;Cell model;Cells;Chronic;Complex;Curcumin;Data;Development;Epithelial;Epithelial Cells;Goals;Growth;Human;IL8 gene;ITGAM gene;Inflammation;Inflammatory Response;Interleukin-6;Knock-out;Lead;Link;Lung;Lung Neoplasms;Macrophage Activation;Malignant neoplasm of lung;Mediating;Membrane;Modeling;Molecular;Mus;Myelogenous;Myeloid Cells;NOD/SCID mouse;Physiologic pulse;Play;Prevention;Preventive;Production;Property;Proteins;Research;Role;SCID Mice;Small Interfering RNA;Structure of parenchyma of lung;TRAF2 gene;Testing;Therapeutic;Tissues;Tumor Necrosis Factor-alpha;Tumor Promotion;Tumor Tissue;Tumor-Derived;Ubiquitination;Urethane;Vascular Endothelial Growth Factors;Vesicle;base;cancer cell;cell growth;clinically relevant;healthy volunteer;human TNF protein;in vivo;innovation;lung Carcinoma;macrophage;migration;mouse model;neoplastic cell;novel;peripheral blood;precursor cell;public health relevance;tumor;tumor growth;ubiquilin

public_health_relevance:
The proposed research is highly relevant to characterizing the association of lung tumor exosomes mediated inflammatory responses and lung tumor growth in that it is focused on characterizing the molecular mechanisms underlying the ability of tumor exosomes to both activate macrophages and promote tumor growth. The central hypothesis is that exosomes produced by lung tumor cells or lung tumor tissue are taken up by bone marrow myeloid precursor cells. These cells further differentiate into macrophages and migrate into the lung promote the progression of urethane- induced lung cancer.


application_id: 8206289
project_number: 5I01BX000216-03
project_title: Nicotine-induced fibronectin expression in lung injury and repair
administering_ic: VA
award_notice_date: 2011-12-06
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000216
project_start_date: 2009-10-01
project_end_date: 2014-03-31
study_section: Respiration[PULM]
contact_pi_person_id: 1876531
contact_pi: ROMAN, JESSE
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2012-04-01
project_abstract:
DESCRIPTION (provided by applicant):     ABSTRACT: Nicotinic acetylcholine receptors (nAChRs) are pentameric channels expressed by neuronal and non-neuronal cells that are capable of signal transduction. Although much of what is known about the function of nAChRs relates to the nervous system information about their roles in non-neuronal systems is rapidly emerging. nAChRs are expressed in lung and interesting work performed in non-human primates suggests that nAChR activation by nicotine leads to pulmonary structural/functional abnormalities. We explored the mechanisms by which nAChR activation leads to matrix expression and found that this effect is mediated through a specific subgroup of nAChRs termed 17 nAChRs. Upon activation of these receptors in lung fibroblasts we observed induction of the MEK-1/Erk pathway and phosphorylation of the transcription factor CREB. In turn CREB stimulated the expression of fibronectin a matrix glycoprotein implicated in lung injury and repair. These events were blocked by 17 nAChR antagonists and were not present in 17 nAChR KO primary lung fibroblasts. In addition we found that nAChR activation stimulates lung fibroblast proliferation through MEK-1/Erk-dependent pathways. Similar observations were made in mice chronically stimulated with nicotine but not in developing and adult 17 nAChR KO animals. More recently we documented an increase in the proliferation and expression of fibronectin in fibroblasts exposed to cigarette smoke extract and observed an increase in fibronectin protein deposition in the lungs of mice exposed to tobacco smoke confirming the relevance of this process to tobacco-related disease. We also found that lung repair mechanisms appeared to be impaired after bleomycin-induced lung injury in the setting of chronic exposure to nicotine. These data unveil a novel role for non-neuronal nAChRs in control of lung tissue remodeling. Based on the above we hypothesize that 17 nAChR activation in lung promotes tissue remodeling by stimulating the proliferation of fibroblasts and their overexpression of matrix components like fibronectin through MEK-1/Erk-dependent signals. At baseline this leads to subtle alterations in matrix composition without causing overt consequences such as fibrosis. In contrast in the setting of lung injury this leads to exaggerated tissue remodeling that promotes inefficient repair and fibrosis. This will be tested in specific aims designed to: 1) Examine the role of 17 nAChRs in the proliferation myofibroblast transdifferentiation and matrix expression of lung fibroblasts and the role of MEK-1/Erk in mediating these events in wildtype vs. 17 nAChR KO lung fibroblasts 2) Examine the role of 17 nAChRs in lung injury and repair using wildtype and 17 nAChR KO mice in two models of lung injury (bleomycin model and the model of heterologous tracheal transplantation) and 3) Examine the effect of persistent 17 nAChR activation in lung repair after injury.    PUBLIC HEALTH RELEVANCE:     Project Narrative This project will explore the cellular mechanisms involved in tissue remodeling of the lung after injury. Tissue remodeling is a process characterized by alterations in the expression deposition and degradation of connective tissue components (also called extracellular matrices). This is important to active combatants but also to veterans in view that many pulmonary disorders typically seen in this population (e.g. COPD lung fibrosis) that are associated with alterations in tissue remodeling. Interestingly tobacco and its components have been implicated in many of these disorders and therefore it is thought that they promote alterations in tissue remodeling. But the mechanisms responsible for these events are unknown. This project will explore the mechanisms involved in these processes by focusing on nicotinic receptors and how activation of these receptors promotes alterations in the relative composition of the lung extracellular matrix.

project_terms:
Adult;Affect;Animals;Bleomycin;Bronchoalveolar Lavage Fluid;Bronchopulmonary Dysplasia;CREB1 gene;Cells;Childhood;Chronic;Chronic Obstructive Airway Disease;Clinical Research;Collagen;Connective Tissue;Data;Deposition;Disease;Event;Exposure to;Extracellular Matrix;Fibroblasts;Fibronectins;Fibrosis;Genes;Genetic Transcription;Glycoproteins;Hamman-Rich syndrome;Harvest;Health;In Vitro;Inbred C57BL Mice;Injury;Ion Channel;Knock-out;Knockout Mice;Link;Lung;Lung diseases;Mediating;Messenger RNA;Modeling;Morphogenesis;Mus;Mutation;Myofibroblast;Neuroglia;Neurons;Nicotine;Nicotinic Receptors;Pathway interactions;Phosphorylation;Play;Population;Process;Production;Promotor (Genetics);Protein Isoforms;Proteins;Receptor Activation;Relative (related person);Reporting;Role;Signal Transduction;Site;Structure of parenchyma of lung;Subgroup;System;Testing;Time;Tissues;Tobacco;Tobacco smoke;Tobacco use;Transplantation;Veterans;Work;alpha-bungarotoxin receptor;base;cigarette smoke;design;drinking water;feeding;human MAP2K1 protein;in vivo;injured;injury and repair;interest;lung development;lung injury;lung repair;mTOR Signaling Pathway;nonhuman primate;novel;outcome forecast;overexpression;prenatal;protein expression;public health relevance;receptor;relating to nervous system;repaired;response;transcription factor;transdifferentiation

public_health_relevance:
Project Narrative This project will explore the cellular mechanisms involved in tissue remodeling of the lung after injury. Tissue remodeling is a process characterized by alterations in the expression deposition and degradation of connective tissue components (also called extracellular matrices). This is important to active combatants but also to veterans in view that many pulmonary disorders typically seen in this population (e.g. COPD lung fibrosis) that are associated with alterations in tissue remodeling. Interestingly tobacco and its components have been implicated in many of these disorders and therefore it is thought that they promote alterations in tissue remodeling. But the mechanisms responsible for these events are unknown. This project will explore the mechanisms involved in these processes by focusing on nicotinic receptors and how activation of these receptors promotes alterations in the relative composition of the lung extracellular matrix.


application_id: 8195631
project_number: 5I01BX000387-02
project_title: Mood-Stabilizing Medications and the Inositol Signaling System
administering_ic: VA
award_notice_date: 2011-04-19
opportunity_number: BX-09-008
application_type: 5
activity_code: I01
ic: BX
serial_number: 000387
project_start_date: 2010-04-01
project_end_date: 2014-03-31
study_section: Neurobiology A[NURA]
contact_pi_person_id: 7118863
contact_pi: PARTHASARATHY, RANGA N
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2011-04-01
project_abstract:
DESCRIPTION (provided by applicant):   
  
Project Summary/Abstract Bipolar disorder (BD) is a devastating condition affecting many Veterans. Lithium and valproate are the drugs of choice for the manic phase of BD. While therapeutically effective their mechanisms of action are not fully understood and undesirable side effects are common. Lithium inhibits brain inositol monophosphatase (IMPase) thereby depleting the available pool of free inositol and resulting in the correction of the manic phase of BD. Inositol synthase (IPS) is a key enzyme that regulates inositol levels by generating the precursor inositol phosphate (IP). Our preliminary data demonstrate that rat brain IPS is up-regulated by chronic lithium treatment which would have the same net effect as inositol depletion relative to inositol phosphate. We hypothesize that lithium and other mood stabilizers exert their therapeutic effects by up-regulating IPS expression increasing the available inositol phosphate pool. This implies that the pathophysiological mechanism underlying BD is a disturbance in maintaining a proper balance in the ratio of free inositol to inositol phosphate pools. Our long-term objective is to define how mood stabilizers work and to find common therapeutic targets in the pathophysiology of BD so new medications can be designed. Our specific aims are: 1) Determine the changes in abundance and activity of the newly-identified isoforms of IPS in brain regions after chronic lithium and valproate; 2) Determine the effects of lithium and valproate on IPS binding to clathrin and receptor-mediated endocytosis; 3) Establish the link between inositol metabolism and behavior in IPS-deficient mice; and 4) Identify drug-targetable convergent pathways for lithium valproate and lamotirgine responsive genes. To accomplish Aim 1 IPS and its isoforms will be quantified in drug-treated rat brain regions by enzyme activity assay Western blot and Q-PCR methods. To accomplish Aim 2 brain IPS and its complexes will be analyzed by Western and mass spectral analyses. Effects on receptor-mediated endocytosis will be monitored by following transferrin receptor trafficking and fluorescence resonance energy transfer (FRET) analysis. For Aim 3 lithium and valproate treated and untreated IPS knock-out mice will be subjected to standardized behavioral tests and their brains assayed for key enzymes in inositol metabolism. Aim 4 will be addressed by microarray and "pathway" analyses to map and compare the chronic lithium valproate and lamotrigine transcriptomes in human neuronal cell lines and in rat primary cortical neuronal cell cultures. Defining the mechanisms of lithium action on inositol metabolism in the brain from the perspective of IPS function may establish a direct connection between inositol regulation and behavior. Thus advancing our understanding of the pathophysiology of BD and disclosing new therapeutic targets for treating BD.

project_terms:
Address;Adverse effects;Affect;Behavior;Behavior Disorders;Behavioral;Binding (Molecular Function);Biochemical;Biological Assay;Biological Process;Bipolar Disorder;Brain;Brain region;Cell Culture Techniques;Cell Line;Cell physiology;Cells;Cerebellum;Chronic;Clathrin;Clinical;Complex;Corpus striatum structure;Data;Drug Delivery Systems;Drug resistance;Endocytosis;Enzymatic Biochemistry;Enzymes;Equilibrium;Fluorescence Resonance Energy Transfer;Foundations;Functional disorder;Future;Gene Expression Profile;Generations;Genes;Global Change;Glucose;Goals;Health;Hippocampus (Brain);Homeostasis;Human;In Vitro;Individual;Inositol;Inositol Metabolism Pathway;Inositol Phosphates;Iraq;Knockout Mice;Link;Lithium;Manic;Manic Disorder;Maps;Measurement;Measures;Mediating;Mental Depression;Messenger RNA;Methods;Modeling;Monitor;Mood stabilizers;Moods;Mus;Mutation;Neurons;Pathway interactions;Patients;Pharmaceutical Preparations;Phospholipids;Phosphorylation;Predisposition;Prefrontal Cortex;Protein Isoforms;Proteins;Psychiatry;RNA;Rattus;Regulation;Relative (related person);Resistance;Route;Services;Signal Pathway;Signal Transduction;System;Testing;Therapeutic;Therapeutic Effect;Time;Transferrin Receptor;Treatment Efficacy;Veterans;War;Western Blotting;Work;abstracting;behavior test;brain pathway;chromatin remodeling;design;diagnostic accuracy;enzyme activity;feeding;gene interaction;improved;lamotrigine;membrane synthesis;mouse model;myo-Inositol-1-Phosphate Synthase;myo-inositol-1 (or 4)-monophosphatase;new therapeutic target;novel;protein protein interaction;receptor mediated endocytosis;research study;response;sugar;therapeutic target;trafficking;valproate

public_health_relevance:
PROJECT NARRATIVE (Potential Relevance to Veterans' Health) Bipolar Disorder (BD) is a major clinical problem for Veterans and there are many Iraq War Veterans treated by VA Psychiatry Service for lithium-resistant BD. To better manage these patients we must be able to identify common targets of mood-stabilizers so that future medications can be designed that have greater efficacy and fewer side effects. One way lithium can correct BD is by modulating brain inositol signal pathways. The brain maintains inositol (a sugar) homeostasis through three routes. Inositol phosphate synthase (IPS) is the only biosynthetic means of generating inositol from glucose. IPS is a new target linked to the genesis of BD. Stimulators of brain IPS enzyme which will then cause the generation of inositol inside the human brain can be considered as an important new class of mood stabilizing medications. The results from these studies will provide a highly positive impact on the health of Veterans with BD.


application_id: 8195630
project_number: 5I01BX000350-03
project_title: Alcohol Proteasome Dysfunction in Liver Injury
administering_ic: VA
award_notice_date: 2011-12-01
opportunity_number: BX-08-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000350
project_start_date: 2009-04-01
project_end_date: 2013-09-30
study_section: Gastroenterology[GAST]
contact_pi_person_id: 1902075
contact_pi: MCCLAIN, CRAIG J.
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2011-10-01
project_abstract:
DESCRIPTION (provided by applicant):     Studies from our VA-supported research and that of others demonstrate a critical role for the proinflammatory cytokine tumor necrosis factor (TNF) in the development and progression of alcoholic liver disease (ALD). Alcoholic liver disease is associated with increased oxidative stress and impaired hepatocyte 26S proteasome function. Our preliminary data demonstrate that inhibition of 26S proteasome function can sensitize hepatocytes to TNF cytotoxicity and higher doses of proteasome inhibitors directly induce hepatocyte death. Additionally this apoptotic death involves the induction of lysosomal membrane permeability (LMP) and is mediated by lysosomal proteases (cathepsins B and D). Based on the increasingly evident role of cathepsins as key downstream mediators of apoptosis and our preliminary results we postulate that chronic alcohol abuse with subsequent proteasome dysfunction leads to LMP and release of lysosomal cathepsins which induce hepatocyte cell death and further sensitize hepatocytes to TNF-mediated hepatotoxicity. Neutrophil infiltration is a hallmark of alcoholic hepatitis and infiltrating neutrophils are thought to play an etiologic role in the liver injury in ALD. We recently demonstrated that inhibition of proteasome function in hepatocytes induces and markedly enhances the TNF-inducible expression of the pro-inflammatory and neutrophil chemotactic cytokine Interleukin-8 (IL-8). It is our working hypothesis that chronic alcohol abuse mediated proteasome dysfunction is associated with increased hepatocyte IL-8 production which significantly contributes to neutrophil infiltration and hepatic injury in ALD. The overall objective of this proposal is to elucidate the cellular and molecular mechanism(s) involved in proteasome dysfunction-mediated hepatic injury associated with ALD. The specific objectives of this proposal are to: 1) Determine the effects of proteasome inhibition and ethanol on sensitization to TNF hepatotoxicity 2) Determine the role of lysosomal membrane permeability and cathepsins B and D in the proteasome dysfunction-mediated hepatocyte apoptosis and sensitization to TNF hepatotoxicity and 3) Determine the effects of proteasome inhibition and ethanol on the hepatocyte expression of the critical neutrophil chemotactic CXC cytokine IL-8. Potential Impact on Veterans Health: Alcoholic liver disease is a leading cause of death from liver disease in the United States and in the Veterans Administration. In studies from the Veterans Administration patients with cirrhosis and superimposed alcoholic hepatitis had > 60% mortality over a 4 year period of time with most of those deaths occurring in the first month. Thus the prognosis for this disease is more ominous than for many common types of cancer such as breast prostate and colon. The longterm goals of this laboratory are to define mechanism(s) for liver cell death in ALD that will provide new insights into this process and provide new areas for therapeutic intervention> This is especially important in ALD where there is no FDA-approved therapy. The potential clinical impact of this work is high.    PUBLIC HEALTH RELEVANCE:      The overall objective of this proposal is to elucidate the cellular and molecular mechanism(s) involved in alcohol and proteasome dysfunction-mediated hepatic injury associated with alcoholic liver disease (ALD). We expect that the results of our study will provide critical molecular insights into the pathogenesis of ALD and provide a basis for clinically relevant paradigms for future development of potential therapeutic intervention(s) in patients with ALD.

project_terms:
26S proteasome;Affect;Alcohol abuse;Alcoholic Hepatitis;Alcoholic Liver Cirrhosis;Alcoholic Liver Diseases;Alcoholic beverage heavy drinker;Alcohols;Animals;Antioxidants;Apoptosis;Apoptotic;Area;Breast;Calpain;Cathepsins;Cathepsins B;Cause of Death;Cell Death;Cell Membrane Permeability;Cells;Cessation of life;Chronic;Cirrhosis;Clinical;Colon;Data;Development;Disease;Dose;Ethanol;Ethanol Metabolism;FDA approved;Functional disorder;Future;Genetic;Goals;Grant;Health;Hepatic;Hepatocyte;Hepatotoxicity;In Vitro;Inflammatory;Injury;Interleukin-8;Label;Laboratories;Liver;Liver diseases;Lysosomes;Mediating;Mediator of activation protein;Mitochondria;Modeling;Molecular;Morbidity - disease rate;Mortality Vital Statistics;Mus;Neutrophil Infiltration;Oxidative Stress;Pathogenesis;Pathway interactions;Patients;Peptide Hydrolases;Permeability;Plasma;Play;Polyubiquitin;Process;Production;Prostate;Proteasome Inhibition;Proteasome Inhibitor;Proteins;Proteolysis;Reactive Oxygen Species;Reduced Glutathione;Research;Research Support;Resistance;Role;Route;Signal Transduction;Stimulus;Therapeutic Intervention;Time;Tumor Necrosis Factor-alpha;Ubiquitin;United States;United States Department of Veterans Affairs;Veterans;Vitamin E;Work;alcohol effect;base;cancer type;chemokine;chronic alcohol ingestion;clinically relevant;cytokine;cytotoxicity;insight;monocyte;multicatalytic endopeptidase complex;neutrophil;outcome forecast;protein degradation;public health relevance;response

public_health_relevance:
The overall objective of this proposal is to elucidate the cellular and molecular mechanism(s) involved in alcohol and proteasome dysfunction-mediated hepatic injury associated with alcoholic liver disease (ALD). We expect that the results of our study will provide critical molecular insights into the pathogenesis of ALD and provide a basis for clinically relevant paradigms for future development of potential therapeutic intervention(s) in patients with ALD.


application_id: 8195629
project_number: 5I01BX000610-03
project_title: Analysis of PTH and Dopamine Receptor Signaling in Proximal Tubules
administering_ic: VA
award_notice_date: 2011-12-01
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000610
project_start_date: 2009-10-01
project_end_date: 2013-09-30
study_section: Nephrology[NEPH]
contact_pi_person_id: 2262379
contact_pi: LEDERER, ELEANOR D
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2011-10-01
project_abstract:
Project Summary/Abstract The goal of this project is to determine how the structural protein NHERF-1 sodium-hydrogenexchanger regulatory factor isoform 1 regulates the trafficking of Npt2a the type IIa sodium phosphatecotransporter to the apical membrane of the renal proximal tubule. The expression of Npt2a at the apicalmembrane is a critical regulatory step because the level of expression and function of Npt2a is the primaryregulator of total body phosphate homeostasis. VA-funded research from this laboratory had revealed that theabsence of NHERF-1 in a model of proximal renal tubule OK (opossum kidney) cells resulted in a markeddecrease in Npt2a expression. The decrease in expression was due to two factors 1) decreased transcriptionof Npt2a and 2) decreased trafficking of Npt2a to the apical membrane. The mechanisms for the faultytrafficking of Npt2a have not been determined and are the subject of this proposal. Npt2a translated in theNHERF-deficient cells (OK-H) lacked a critical post-translational modification glycosylation. These proteinsinstead of trafficking to apical membrane accumulated in a perinuclear location. Inefficient apical membranelocalization had previously been described in a mouse lacking expression of NHERF-1. Expression of aNHERF-1 construct that lacked a normal PDZ-2 domain did not allow trafficking of Npt2a to the apicalmembrane. NHERF-1 is a multiple PDZ domain protein which interacts with multiple signaling molecules andtransporter proteins. A role for the PDZ-2 domain in Npt2a regulation had not been observed previously.Finally inhibition of SNARE (SNAP Receptor) protein interaction using a competing peptide introduced into OKcells blocked insertion of Npt2a into the apical membrane demonstrating that Npt2a is transported to theapical membrane via a vesicular transport mechanism. The preliminary data suggested the hypothesis that NHERF-1 is essential for the forward trafficking ofNpt2a from site of synthesis to the apical membrane. First the role of the post-translational modificationsglycosylation and phosphorylation on Npt2a apical membrane trafficking will be examined by determining thecellular localization of transfected Npt2a constructs lacking the motifs required for these post translationalmodifications in a cell culture model. The glycosylation phosphorylation and ubiquitination states of Npt2a willbe compared in NHERF-replete and NHERF-deficient OK cells and in wild type and NHERF-1 knock outmouse proximal tubule cells. The effect of inhibitors of glycosylation on the intracellular localization of Npt2awill be determined. Second the intracellular site or sites of interaction between NHERF-1 and Npt2a will beanalyzed. Forward trafficking of Npt2a labeled by a GFP tag and by S35 methionine in OK cells and OK-H cellswill be slowed by culture at 20 C. Analysis of Npt2a localization by confocal imaging density centrifugationand immunoelectronmicroscopy will be performed at sequential time points until the proteins are detected asbiotinylated forms indicating appropriate insertion into the apical membrane. Trafficking of Npt2a in the twocell culture models will be compared. Proteins associated with Npt2a at each time point will be determined byimmunoprecipitation and proteomic analysis. To define which steps in the forward trafficking are NHERF-dependent Npt2a trafficking will be compared in NHERF-replete and NHERF-deficient cells under conditionsof traffic arrest: ezrin deficiency inhibition of SNARE interaction and CLC-5 (intracellular chloride channelCLC family isoform 5) deficiency. Third the sites on Npt2a and NHERF-1 critical for the NHERF-1 effect onNpt2a trafficking will be determined by mutational analysis of both proteins followed by both in vitro analysis ofprotein interaction and assessment of intracellular interaction by FRET methodology. These studies will define where NHERF-1 acts in Npt2a forward trafficking define the sites on bothproteins responsible for their interaction and yield mechanistic insights for this unique functional process.

project_terms:
Address;Affect;Age;Apical;CLC Gene;Cardiovascular Diseases;Carrier Proteins;Cell Culture Techniques;Cell Line;Cells;Centrifugation;Chloride Channels;Country;Data;Didelphidae;Disease;Dopamine Receptor;Event;Exhibits;Family;Fluorescence Resonance Energy Transfer;Funding;Genetic Transcription;Goals;Health;Hip Fractures;Homeostasis;Image;Immunoprecipitation;In Vitro;Individual;Kidney;Kidney Calculi;Knockout Mice;Label;Laboratories;Laboratory Research;Location;Medical;Membrane Microdomains;Membrane Protein Traffic;Membrane Transport Proteins;Metabolic Bone Diseases;Methionine;Methodology;Modeling;Molecular;Morbidity - disease rate;Mortality Vital Statistics;Mus;Mutation;Pathway interactions;Peptides;Phosphorylation;Physiologic pulse;Play;Population;Post-Translational Protein Processing;Process;Protein Analysis;Protein Isoforms;Proteins;Proteomics;Proximal Kidney Tubules;Receptor Signaling;Regulation;Renal function;Research;Research Personnel;Role;SNAP receptor;Signaling Molecule;Site;Staging;Structural Protein;Tertiary Protein Structure;Time;TimeLine;Transfection;Translating;Transport Vesicles;Tubular formation;Ubiquitination;Vesicle;Veterans;Work;abstracting;apical membrane;base;cardiovascular risk factor;caveolin 1;cold temperature;density;design;ezrin;glycosylation;in vitro Assay;inhibitor/antagonist;inorganic phosphate;insight;kidney cell;mutant;protein expression;receptor;research study;sodium-hydrogen exchanger regulatory factor;sodium-phosphate cotransporter proteins;solute;syntaxin binding protein 1;trafficking;wasting

public_health_relevance:
Project Narrative  The goal of this project is to determine how the protein NHERF-1 sodium-hydrogen exchanger regulatory factor isoform 1 regulates the trafficking of Npt2a the type IIa sodium phosphate cotransporter to the apical membrane of the renal proximal tubule. The kidney regulates the total body content of phosphate through altering the expression of Npt2a at the apical membrane. Phosphate wasting can cause kidney stones and metabolic bone disease while phosphate retention is a risk factor for cardiovascular events common diseases in Veterans of all ages. Cardiovascular disease is the major cause of morbidity and mortality in the Veteran population. Kidney stones tend to affect individuals during their most productive years resulting in over $2 billion dollars of medical expenses in this country. Metabolic bone disease contributes substantially to the occurrence of hip fracture a disabling event with up to a 50% one year mortality. Thus understanding how the kidney regulates phosphate homeostasis has significant relevance to the health of Veterans of all ages.


application_id: 8195617
project_number: 5I01BX000672-03
project_title: Class B Scavenger Receptors and Atherosclerosis
administering_ic: VA
award_notice_date: 2011-12-01
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000672
project_start_date: 2009-10-01
project_end_date: 2013-09-30
study_section: Cardiovascular Studies A[CARA]
contact_pi_person_id: 7875298
contact_pi: VAN DER WESTHUYZEN, DENEYS REM
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2011-10-01
project_abstract:
The roles of the Class B scavenger receptors SR-BI and CD36 in the development ofatherosclerotic disease a major cause of death remain poorly understood. This is due in part to thevaried and still little understood functions of these two pattern recognition receptors in host defenseand tissue homeostasis. The role of CD36 in the initiation of macrophage foam cell formation iscontroversial although CD36 has the ability to take up modified lipoproteins such as oxidized LDL.Other potentially important functions of CD36 include its role in the innate immune system and itsrole in inflammatory signaling. Atherosclerosis is a chronic inflammatory disease and we proposethat CD36 plays a more complex role in atherosclerotic lesion formation. The structurally closelyrelated receptor SR-BI plays a key role in reverse cholesterol transport to the liver and has beenshown in several mouse models to be anti-atherogenic. Recent findings have also indicatedadditional extrahepatic roles of SR-BI in atherogenesis including effects in macrophages. Thefunctional basis for these effects is not known although we have shown that SR-BI-null mice have amarkedly enhanced inflammatory cytokine response and higher lethality in response tolipopolysaccharide (LPS). Furthermore mice deficient in both SR-BI and CD36 unexpectedly exhibitmuch greater atherosclerotic lesion formation than mice lacking either SR-BI or CD36 only. Thesefindings indicate that SR-BI and CD36 each have athero-protective functions yet to be defined. Theprotective functions of the two receptors may be overlapping allowing at least partial compensationin mice lacking only one of the two receptors. The central hypothesis of this proposal is that SR-BIand CD36 act in a complementary or synergistic manner to protect against atherosclerosis. Wepropose that SR-BI and CD36 are key to the innate defense against atherosclerosis a chronicinflammatory disease. The following 4 objectives are proposed to examine these hypotheses:Objective 1 will examine the complementary roles SR-BI and CD36 in atherosclerotic lesionformation in response to high fat diets. This will be accomplished by analyzing the extent and natureof aortic lesions in wild type SR-BI-null CD36-null and SR-BI-nullXCD36-null mice. Objective 2 willinvestigate the roles of macrophage-expressed Class B scavenger receptors in atherosclerosis. Thiswill be accomplished through the use of bone marrow transplantation. Objective 3 will identify thecomplementary or synergistic roles of SR-B1 and CD36 in macrophage foam cell formation. This willbe accomplished by measuring lipid accumulation in wild type and SR-BI-XCD36- DKOmacrophages treated with modified LDLs. Cells will be examined for lipid related protein expressionand by gene microarray analysis. Lipid uptake and efflux pathways will be compared. Objective 4 willinvestigate enhanced inflammatory signaling in CD36/SR-BI DKO macrophages and its effects onmacrophage lipid accumulation. The impact of inflammatory signaling on modified LDL uptake inprimary macrophages will be evaluated together with the hypothesis that increased plasmamembrane cholesterol content in SR-BIxCD36 DKO macrophages stimulates inflammatory signalingpathways. These studies are expected to provide new understanding of the metabolic functions ofCD36 and SR-BI as well as their role in atherosclerosis.

project_terms:
Affect;Antiatherogenic;Arterial Fatty Streak;Arteries;Atherosclerosis;Bone Marrow Transplantation;CD36 gene;Cardiovascular Diseases;Cause of Death;Cell membrane;Cells;Cholates;Cholesterol;Chronic;Complex;Development;Diet;Disease;Exhibits;Extrahepatic;Fatty acid glycerol esters;Financial compensation;Foam Cells;Genes;Hepatic;Homeostasis;Host Defense;Immune system;Inflammation;Inflammatory;Inflammatory Response Pathway;Knock-out;Knockout Mice;LDL Cholesterol Lipoproteins;Lesion;Lipids;Lipopolysaccharides;Lipoprotein Receptor;Lipoproteins;Liver;Low-Density Lipoproteins;Measures;Metabolic;Metabolism;Microarray Analysis;Mus;Nature;Pathway interactions;Pattern recognition receptor;Plasma;Play;Regulation;Risk Factors;Role;SR-B proteins;SR-BI receptor;Signal Pathway;Signal Transduction;TLR4 gene;Testing;Tissues;Transplantation;United States;Veterans;atherogenesis;base;drug development;feeding;macrophage;mouse model;oxidized low density lipoprotein;protein expression;public health relevance;receptor;research study;response;reverse cholesterol transport;scavenger receptor;treatment strategy;uptake

public_health_relevance:
Cardiovascular disease remains the number one killer in the United States and among veterans. Although widely studied the mechanisms underlying atherosclerosis are not yet clearly understood. Plasma cholesterol and LDL levels are a major risk factor for atherosclerosis but the underlying mechanisms for this are still poorly understood. Further understanding of how LDL cholesterol accumulates in the artery wall and the roles of the various receptors for lipoproteins will potentially allow for the identification of new treatment strategies and new drug development.


application_id: 8195616
project_number: 5I01BX000325-03
project_title: Studies on the role of IL-23p19/IL-17 cascade in the pathogenesis of acute GVHD
administering_ic: VA
award_notice_date: 2011-12-06
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000325
project_start_date: 2009-10-01
project_end_date: 2012-12-31
study_section: Immunology and Dermatology A[IMMA]
contact_pi_person_id: 8781506
contact_pi: THOMPSON, JOHN SECORD
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2012-01-01
project_abstract:
DESCRIPTION (provided by applicant):     Abstract It is firmly established that aGVHD is initiated by T-cell recognition of antigenic foreignness and ex vivo depletion of T-cells has been the most effective method to reduce its incidence and severity. The proposed studies are to test 2 hyptheses: 1) The IL-23p19/IL-17 cytokine axis plays a major role in the inflammatory reactions characteristic of aGVHD by regulating the expression of key cytokine genes that may promote or inhibit aGVHD. 2) The IL -23p19/1L-17 cytokine axis interacts closely but not completely with the TNF1 IL-12 and IL-6 cascade to amplify the pathology of aGVHD. Three objectives are designed to address these They are derived from our observation that BALB/c mice transplanted with bone marrow (BM) plus T- containing spleens (SCs) from mutant mice deficient in the p19 dimer of the IL-23 p19/p40 heterodimer develop aGVHD less frequently and less severely than BALB/c transplanted with normal wild type C57BL/6 BM + SCs. IL-17 mRNA and serum cytokine produced by CD8 Th17 cells are elevated in BALB/c mice transplanted with both WT and p19 deficient cells but to significantly higher levels in the WT animals. Induction of Th17 producing IL-17 in the p19-/- mice was shown to be derived from TGF2 IL-6 and IL23p19 induced by total body irradiation of the host BALB/c animals. The first objective is to determine the effect on the induction of aGVHD by isolated T and other cells in the donor inoculum. The second objective is to determine the role of the products of Th17 cells in the pathogenesis of aGVHD. The third objective is to determine the relative roles of the IL-23/IL17-21-22 cascade versus the TNF1-IL-12-IL-6 cascade in the pathology of aGVHD. Use of target mutant animals deficient in IL-23p19 RORgamma t (critical for Th17 cell development) and TNF1 provide unique models in which to dissect the components of aGVHD pathology.    PUBLIC HEALTH RELEVANCE:     Relevance to VA Mission: Every day large numbers of unfortunate veterans present with new cancers. In addition to the leukemias myelodysplasia and multiple myeloma other cancers could be amenable to stem cell transplantation. Admittedly there are some cancers that probably would not benefit from this therapy but the ability to achieve a revitalized immune system could be an important addition to their care. However as stated above the current risk of developing transplant related toxicities GVHD and overwhelming infection are too great to offer stem cell transplantation to patents with a wider variety of cancers. The proposed studies are to investigate a new approach to minimize GVHD

project_terms:
Acute;Acute Graft Versus Host Disease;Address;Allogenic;Animals;Bone Marrow;CD8B1 gene;Caring;Cells;Characteristics;Chronic Graft Versus Host Disease;Cytokine Gene;Development;Disease;Dysmyelopoietic Syndromes;Graft-vs-Host Disease;Health;IL-23 p19;IL17 gene;Immune system;Inbred BALB C Mice;Incidence;Infection;Inflammatory;Interleukin-12;Interleukin-17;Interleukin-6;Legal patent;Malignant Neoplasms;Messenger RNA;Methods;Mission;Modeling;Multiple Myeloma;Mus;Mutant Strains Mice;Pathogenesis;Pathology;Play;Reaction;Relative (related person);Risk;Role;Serum;Severities;Spleen;Stem cell transplant;Survivors;T-Cell Depletion;T-Lymphocyte;Testing;Toxic effect;Transplantation;Veterans;Whole-Body Irradiation;abstracting;cytokine;design;dimer;experience;interleukin-23;leukemia;mutant;novel strategies

public_health_relevance:
Relevance to VA Mission: Every day large numbers of unfortunate veterans present with new cancers. In addition to the leukemias myelodysplasia and multiple myeloma other cancers could be amenable to stem cell transplantation. Admittedly there are some cancers that probably would not benefit from this therapy but the ability to achieve a revitalized immune system could be an important addition to their care. However as stated above the current risk of developing transplant related toxicities GVHD and overwhelming infection are too great to offer stem cell transplantation to patents with a wider variety of cancers. The proposed studies are to investigate a new approach to minimize GVHD


application_id: 8195615
project_number: 5I01BX000622-03
project_title: Mechanisms of Renal Lipid Accumulation in Diabetic Nephropathy
administering_ic: VA
award_notice_date: 2011-12-01
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000622
project_start_date: 2009-10-01
project_end_date: 2013-09-30
study_section: Nephrology[NEPH]
contact_pi_person_id: 3123430
contact_pi: TANNOCK, LISA R
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2011-10-01
project_abstract:
DESCRIPTION (provided by applicant):     Nephropathy is a major complication of both type 1 and type 2 diabetes which causes considerable morbidity and mortality. Despite best current treatments significant numbers of individuals with albuminuria progress to end stage renal disease. Although the mechanisms underlying this progression are not fully understood renal lipid and lipoprotein accumulation has been shown to accelerate the development of nephropathy. Renal lipid accumulation triggers an influx of inflammatory cells with subsequent development of glomerulosclerosis the characteristic lesion of diabetic nephropathy. Glomerulosclerosis is characterized by increased deposition of mesangial matrix including proteoglycans. Of particular interest the renal content of the small leucine-rich proteoglycan (SLRP) biglycan is increased in diabetes. TGF-2 which is elevated in diabetes and is known to be a key mediator of diabetic nephropathy development and progression also increases mesangial matrix deposition including increased expression of biglycan. Furthermore we have shown that TGF-2 increases the size and LDL binding affinity of renal proteoglycans. Thus increased renal proteoglycan (biglycan) synthesis induced by elevated TGF-2 in diabetes may be responsible for mediating the renal accumulation of lipoproteins. The overall goal of this grant is to test the hypothesis that renal lipid accumulation is mediated through interactions of lipoproteins with renal proteoglycans especially biglycan. This will be tested by comparing diabetic nephropathy in mice expressing proteoglycan-binding defective LDL with littermates expressing wildtype LDL. To determine if biglycan is the key proteoglycan responsible diabetic nephropathy will be compared between biglycan deficient and wildtype mice. The experiments outlined in this grant will provide direct in vivo experimental data identifying if proteoglycan mediated renal lipid accumulation contributes significantly to the development and progression of diabetic nephropathy and if the key proteoglycan is biglycan. This grant will also identify if biglycan serves as a natural inhibitor of TGF-2 in vivo. Thus the significance of this proposal is that it not only identifies a mechanism linking hyperlipidemia and diabetic nephropathy but also will identify novel targets to prevent or intervene in the development of diabetic nephropathy.    PUBLIC HEALTH RELEVANCE:     Potential Impact on Veterans Health Care: Veterans have very high prevalence of diabetes hyperlipidemia and diabetes complications including nephropathy. This research will either identify or exclude proteoglycans as a cause of renal lipid accumulation contributing to diabetic nephropathy. The grant will prove the concept in an animal model system and the results can be easily translated into clinical research. The ultimate goal is to identify novel targets for therapeutic intervention to decrease the morbidity and mortality of diabetic nephropathy.

project_terms:
Affinity;Albuminuria;Animal Model;Antibodies;Binding (Molecular Function);Biological Models;Cells;Characteristics;Clinical Research;Complication;Complications of Diabetes Mellitus;Data;Deposition;Development;Diabetes Mellitus;Diabetic Nephropathy;Enalapril;End stage renal failure;Goals;Grant;Healthcare;High Prevalence;Hyperlipidemia;In Vitro;Individual;Inflammatory;Kidney;Kidney Diseases;Lesion;Leucine;Link;Lipids;Lipoproteins;Low Density Lipoprotein Receptor;Low-Density Lipoproteins;Mediating;Mediator of activation protein;Modeling;Morbidity - disease rate;Mortality Vital Statistics;Mus;Proteoglycan;Reporting;Research;Role;Testing;Therapeutic Intervention;Tissues;Transforming Growth Factor beta;Translating;Up-Regulation (Physiology);Veterans;biglycan;diabetic;glomerulosclerosis;in vivo;inhibitor/antagonist;interest;mouse model;nephrogenesis;neutralizing antibody;novel;prevent;public health relevance;receptor binding;research study;type I and type II diabetes

public_health_relevance:
Potential Impact on Veterans Health Care: Veterans have very high prevalence of diabetes hyperlipidemia and diabetes complications including nephropathy. This research will either identify or exclude proteoglycans as a cause of renal lipid accumulation contributing to diabetic nephropathy. The grant will prove the concept in an animal model system and the results can be easily translated into clinical research. The ultimate goal is to identify novel targets for therapeutic intervention to decrease the morbidity and mortality of diabetic nephropathy.


application_id: 8181319
project_number: 5I01RX000338-02
project_title: TBI epileptogenesis: pathologic hippocampal L-glut synaptic plasticity
administering_ic: VA
award_notice_date: 2014-04-14
opportunity_number: RX-10-005
application_type: 5
activity_code: I01
ic: RX
serial_number: 000338
project_start_date: 2010-10-01
project_end_date: 2014-09-30
study_section: Brain Health and Injury[RRD1]
contact_pi_person_id: 9405862
contact_pi: SLEVIN, JOHN T.
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2011-10-01
project_abstract:
DESCRIPTION (provided by applicant):   
  
Project Summary The molecular mechanism that underlies evolution to post-traumatic epilepsy (PTE) in individuals with traumatic brain injury (TBI) is unknown. It has been suggested that the development of PTE following TBI is the combined result of kindling epileptogenesis during the early stages of recovery and triggering by residual irritation from damaged neural tissue. Glutamate the major excitatory neurotransmitter (NT) in the central nervous system must be tightly regulated to allow for proper neuronal signaling without creating excessive excitation. TBI produces an increase in glutamate release into the extracellular space and a concurrent decrease in the ability of excitatory amino acid transporters to reuptake glutamate that may lead to its accumulation propagating secondary damage cascades and producing aberrant neuronal signaling. We suggest that the enhanced hippocampal glutamate levels seen post-TBI are in part due to changes of the neurosecretory machinery and SNARE regulators and/or re-sequestration process. Left unchecked these changes become drivers of post-TBI epileptogenesis leading to PTE. The hypothesis that pathologic adaptation of inherent hippocampal glutamatergic synaptic plasticity promotes post-TBI epileptogenesis will be tested through three specific aims in animals (and appropriate controls) that have been administered a lateral fluid percussion injury (LFPI) and then monitored by telemetry until electroencephalographic (EEG) evidence of epileptiform activity: (1) Measure of tonic glutamate levels and evoked glutamateamate release in the hippocampal formation of epileptic and non-epileptic animals after moderate LFPI through use of a novel microarray electrode system. In tissue from hippocampal formation of these same animals we will use Western blotting techniques to quantify (2) components of the neurotransmitter release process and (3) certain glutamate transporters. Investigation of this putative relationship between aberrant glutamate neuronal exocytosis/resequestration and PTE will provide insight into potential mechanisms by which glutamate neurotransmission is regulated and into the molecular events of epileptogenesis as well as provide potential foci for therapeutic intervention. The long-term goal of this project is to restore to normal aberrant mechanisms associated with TBI-associated epileptogenesis. There are indications from animal studies that several newer FDA-approved and actively used antiepileptic drugs e.g. levetiracetam and lacosamide may also act as antiepileptogenic agents through effect on glutamate neurotransmission. Such agents would be appropriate compounds for further mechanistic evaluation in animals and clinical studies in PTE patients if the studies proposed here suggest that chronic dysregulation of glutamatergic release is associated with epileptogenesis. Pretransfection with a recently developed adeno-associated virus for the GLT-1 transporter to enhance glutamate reuptake currently under investigation by one of us may provide yet another novel approach to target a specific mechanism involved in glutamate regulation should these studies confirm a glutamate transporter abnormality is associated with development of PTE.

project_terms:
Acute;Affect;Afghanistan;Amino Acid Transporter;Amygdaloid structure;Animal Model;Animals;Antiepileptic Agents;Antiepileptogenic;Behavior;Chronic;Clinical Research;Dependovirus;Development;Electrodes;Enzymes;Epilepsy;Epileptogenesis;Evaluation;Event;Evolution;Excitatory Amino Acids;Exocytosis;Extracellular Space;FDA approved;Glutamate Transporter;Glutamates;Goals;Healthcare Systems;Hippocampal Formation;Hippocampus (Brain);Human;Individual;Injury;Investigation;Iraq;Kindling (Neurology);Label;Lateral;Lead;Left;Levetiracetam;Life;Measures;Microelectrodes;Military Personnel;Modeling;Molecular;Molecular Target;Monitor;Morbidity - disease rate;Mortality Vital Statistics;Neuraxis;Neurons;Neurotransmitters;Pathologic;Patients;Population;Post-Traumatic Epilepsy;Probability;Process;Recovery;Regulation;Research;Residual state;Risk Factors;SNAP receptor;Signal Transduction;Site;Soldier;Staging;Survivors;Synapses;Synaptic plasticity;System;Techniques;Telemetry;Testing;Therapeutic;Therapeutic Intervention;Tissues;Traumatic Brain Injury;United States;United States Department of Veterans Affairs;Veterans;Western Blotting;base;combat;dentate gyrus;economic cost;fluid percussion injury;insight;irritation;neurotransmission;neurotransmitter release;neurotransmitter reuptake;novel;novel strategies;presynaptic;relating to nervous system;reuptake;therapeutic target;uptake

public_health_relevance:
In addition to affecting civilian populations traumatic brain injury (TBI) has been labeled the signature injury of the current combat in Iraq and Afghanistan. TBI is a leading cause of morbidity and mortality in the United States and is a risk factor for post-traumatic epilepsy (PTE). Examining TBI-induced dysregulation of glutamate neurotransmitter release and reuptake the specific aims of this proposal may reveal important mechanisms identify potential therapeutic targets and pinpoint the therapeutic window to eliminate or reduce the probability of developing PTE. Recognition of molecular targets for new therapies that would extinguish TBI- induced epileptogenesis would alleviate the human suffering of civilians soldiers and veterans as they return to civilian life as well as diminish the concomitant long-term economic costs of epilepsy to the US health care system the military and the Veterans Administration.


application_id: 8141677
project_number: 1I01BX001204-01
project_title: Renoprotective effect of increased PKG activity in diabetic nephropathy
administering_ic: VA
award_notice_date: 2011-06-21
opportunity_number: BX-10-006
application_type: 1
activity_code: I01
ic: BX
serial_number: 001204
project_start_date: 2011-07-01
project_end_date: 2015-06-30
study_section: Nephrology[NEPH]
contact_pi_person_id: 7837422
contact_pi: WANG, SHUXIA
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2011-07-01
project_abstract:
DESCRIPTION (provided by applicant):   
  
Diabetic nephropathy is the single most common cause of end-stage renal failure and is highly prevalent in both type 1 and type 2 diabetics. Hyperglycemia has been suggested to play a critical role in the pathogenesis of diabetic nephropathy. Transforming growth factor-2 (TGF-2) is a major effector leading to renal fibrosis in response to hyperglycemia. TGF-2 is expressed as a biologically inactive form and must be converted to its active form to elicit fibrogenic effects. Thrombospondin1 (TSP1) has been identified as the molecular regulator of TGF-2 activation under diabetic conditions. TSP1-mediated TGF-2 activation is involved in the development of experimental diabetic nephropathy and also plays a role in human diabetic nephropathy. Preliminary data demonstrate a novel inhibitory effect of cGMP-dependent protein kinase (PKG) on glucose-induced TSP1 expression and TSP1-mediated TGF-2 activation in mesangial cells suggesting a therapeutic potential of PKG in diabetic nephropathy. The proposed studies will test whether genetically or pharmacologically increased PKG activity prevents the development of diabetic nephropathy through inhibition of TSP1-mediated TGF-2 activation in different diabetic mouse models. Specific Aim 1 will determine the mechanisms by which high glucose down-regulates PKG activity in kidney cells in vitro and its role in glucose-induced TSP1 expression TGF-2 levels/activity and extracellular matrix (ECM) production. Specific Aim 2 will determine whether increased PKG activity prevents the development of diabetic nephropathy in vivo. These studies will establish the significance of PKG in the development of diabetic nephropathy. Importantly utilization of sildenafil (an inhibitor of cGMP specific phosphodiesterase 5) to increase PKG activity in the proposed studies is translational and relevant to human therapy. Sildenafil is a drug used to treat erectile dysfunction (ED another diabetic complication in male patients) and pulmonary arterial hypertension (PAH) in clinic. The current proposed studies will test a novel potential application of sildenafil in the treatment of diabetic nephropathy in different diabetic mouse models. Therefore these studies have significant clinical relevance and will lead to the development of novel therapies to attenuate/treat diabetic nephropathy a major complication of diabetes.  
  
PUBLIC HEALTH RELEVANCE:   
  
More than 20% of veterans or one million veterans who use VA health care system are affected by diabetes. Veterans also have very high prevalence of diabetic complications including nephropathy. Nephropathy is a major complication of diabetes which leads to considerable morbidity and mortality. Despite current improved treatments significant numbers of patients with albuminuria progress to end stage renal disease which requires dialysis or kidney transplantation. This proposal aims at defining the novel roles for PKG a cGMP- dependent protein kinase in the development of diabetic nephropathy and testing the novel therapeutic potential of genetically or pharmacologically increased PKG activity in attenuating diabetic nephropathy in different mouse models of diabetes. The success of this proposal will benefit the veterans' health care: improving the quality of life of veterans and reducing the health care expenditures in the VA health care system.


application_id: 8044373
project_number: 1I01BX000962-01
project_title: Exosomal TRAF2-CSN5 complex mediated inflammation promotes tumor growth
administering_ic: VA
award_notice_date: 2011-01-18
opportunity_number: BX-10-001
application_type: 1
activity_code: I01
ic: BX
serial_number: 000962
project_start_date: 2011-04-01
project_end_date: 2015-03-31
study_section: Immunology and Dermatology A[IMMA]
contact_pi_person_id: 1889248
contact_pi: ZHANG, HUANG-GE
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2011-04-01
project_abstract:
DESCRIPTION (provided by applicant):   
  
The overall goal of this proposal is to elucidate the mechanisms by which lung tumor exosomes promote an inflammatory response namely the activation of macrophages and promote the progression and growth of lung tumors. Our preliminary data indicate that pretreatment of A/J mice with exosomes produced by TC-1 lung tumor cells or exosomes isolated from the lung tissue of A/J mice that have been pretreated with the carcinogen urethane results in more rapid tumor growth and earlier progression of lung carcinomas. Urethane treatment results in the recruitment of the activated form of TRAF2 to the exosomes and the exosomes with activated TRAF2 are taken up by the bone marrow-derived precursors of macrophages leading to their maturation and subsequent migration to the lung. The recruitment of the activated TRAF2 to the exosomes requires degradation of the inflammation suppressor protein CYLD in the tumor cells; moreover TRAF2 can mediate ubiquination of CYLD thereby promoting its degradation. The results of siRNA TRAF2 knockout indicated however that TRAF2 is required but is not sufficient for ubiquitination of CYLD. We have now identified two exosomal proteins that interact with TRAF2 Itch and CSN5 which have the potential to enhance the ubiquitination of CYLD in lung tumor cells. Our recent data indicating that CYLD is not packed in exosomes isolated from the peripheral blood of lung cancer patients although it is present in exosomes from healthy volunteers indicate the potential clinical relevance of this model that directly links the effects of a carcinogen with an inflammatory response and promotion of tumor development. We propose to: (1) Confirm the role of lung tumor exosomal TRAF2 in the promotion of urethane-induced lung carcinomas by determining whether knockout of TRAF2 in TC-1 lung tumor cells is sufficient for (i) inhibition of macrophage differentiation and (ii) prevention of lung tumor exosome-mediated enhancement of urethane induced lung cancer. (2) Determine if other exosomal proteins that interact with CYLD participate in the ubiquitination of CYLD in TC-1 tumor cells and determine if the elimination of these proteins stabilizes exosomal CYLD and suppresses exosomal TRAF2 activation resulting in the inhibition of macrophage activation and prevention of tumor growth. (3) Determine if macrophages pulsed with lung tumor exosomes isolated from lung cancer patients promote lung tumor growth in a NOD-SCID mouse model. The data generated should permit the development of a highly innovative model of the cellular and molecular basis for exosome-mediated chronic inflammation and promotion of tumor growth and indicate novel preventive and therapeutic strategies.  
  
PUBLIC HEALTH RELEVANCE:   
  
The proposed research is highly relevant to characterizing the association of lung tumor exosomes mediated inflammatory responses and lung tumor growth in that it is focused on characterizing the molecular mechanisms underlying the ability of tumor exosomes to both activate macrophages and promote tumor growth. The central hypothesis is that exosomes produced by lung tumor cells or lung tumor tissue are taken up by bone marrow myeloid precursor cells. These cells further differentiate into macrophages and migrate into the lung promote the progression of urethane- induced lung cancer.

project_terms:
A/J Mouse;Acceleration;Anti-Inflammatory Agents;Bone Marrow;Bone Marrow Cells;Cancer Patient;Carcinogens;Cell model;Cells;Chronic;Complex;Curcumin;Data;Development;Epithelial;Epithelial Cells;Goals;Growth;Human;IL8 gene;ITGAM gene;Inflammation;Inflammatory Response;Interleukin-6;Knock-out;Lead;Link;Lung;Lung Neoplasms;Macrophage Activation;Malignant neoplasm of lung;Mediating;Membrane;Modeling;Molecular;Mus;Myelogenous;Myeloid Cells;NOD/SCID mouse;Physiologic pulse;Play;Prevention;Preventive;Production;Property;Proteins;Research;Role;SCID Mice;Small Interfering RNA;Structure of parenchyma of lung;TRAF2 gene;Testing;Therapeutic;Tissues;Tumor Necrosis Factor-alpha;Tumor Promotion;Tumor Tissue;Tumor-Derived;Ubiquitination;Urethane;Vascular Endothelial Growth Factors;Vesicle;base;cancer cell;cell growth;clinically relevant;healthy volunteer;human TNF protein;in vivo;innovation;lung Carcinoma;macrophage;migration;mouse model;neoplastic cell;novel;peripheral blood;precursor cell;tumor;tumor growth;ubiquilin


application_id: 8044269
project_number: 1I01BX000693-01A2
project_title: Airway sensors with myelinated afferents
administering_ic: VA
award_notice_date: 2010-12-02
opportunity_number: BX-10-001
application_type: 1
activity_code: I01
ic: BX
serial_number: 000693
suffix: A2
project_start_date: 2010-10-01
project_end_date: 2014-09-30
study_section: Respiration[PULM]
contact_pi_person_id: 2084836
contact_pi: YU, JERRY (JUN)
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
project_abstract:
DESCRIPTION (provided by applicant):   
  
The long-term objective of this proposal is to identify the unknown morphology and pathophysiological functions of pulmonary sensory receptors potentially benefiting more than 30 million patients with chronic obstructive pulmonary disease (COPD) and other pulmonary diseases in the US including veterans. Dyspnea bronchial inflammation hyperreactivity coughing and mucus secretion occur when airway sensory receptors are activated. These are common clinical manifestations of pulmonary disorders; however the underlying neural mechanisms remain unclear in part because our knowledge of airway receptor morphology and sensory behavior is limited. Increasing evidence points to important roles of myelinated airway receptors in pulmonary diseases. In this application we will test the hypothesis that vagal airway sensory units are not only transducers but also processors. Sensory units' behaviors depend on their structures. In addition a single unit may contain homogeneous or heterogeneous types of receptors providing varied mixed behavior. In this application we will address three specific aims to characterize the morphology and sensory behavior of myelinated airway receptors. 1) To identify different morphological structures of myelinated airway receptors by examing receptor morphology by immunohistochemical or neural tracing techniques combined with confocal microscopy. 2) To assign a specific receptor structure to physiologically defined classes of receptors by first electrophysiologically characterizing each known receptor type then tissue in the receptive field will be isolated dissected and immunohistochemically stained for receptor morphology. 3) To explore mechanosensor structure-function relationships and sensory integration mechansims by examining sensory behavior before and after blocking or stimulating one of the receptors in the unit. Thus integration mechanisms of sensory units can be explored. Successful completion of these studies will provide important structure-function relationships and establish a comprehensive objective classification system for the airway receptors. The presence of heterogenous receptors challenges the current view of the airway sensory unit. Transmission of heterogeneous information in a single unit verifies that the unit is not merely a transducer but also a processor. Our data will lay the foundation for future research and facilitate our understanding of the receptor function and their roles in pulmonary diseases and may help improve managements for cardiopulmonary patients suffering from cardiac failure COPD asthma acute lung injury and other pulmonary diseases.  
  
PUBLIC HEALTH RELEVANCE:   
  
Airway sensory receptors play important roles in cardiopulmonary diseases. Understanding their mechanisms of operation may help patient management thus benefitting a large number of veterans who suffer from cardiac and lung diseases including cardiac failure acute respiratory distress syndrome (ARDS) chronic obstructive pulmonary disease (COPD) and asthma.

project_terms:
Address;Adult Respiratory Distress Syndrome;Asthma;Behavior;Bronchoconstriction;C Fiber;Cardiopulmonary;Chemicals;Chronic Obstructive Airway Disease;Classification;Clinical;Confocal Microscopy;Cor pulmonale;Coughing;Data;Development;Disease;Dyspnea;Exhibits;Fiber;Foundations;Heart Diseases;Heart failure;Inflammation;Knowledge;Laboratories;Lung;Lung Injury Acute;Lung diseases;Mechanics;Mechanoreceptors;Microscopy;Morphology;Mucous body substance;Neurons;Patients;Physiological;Play;Process;Research;Role;Sensory;Sensory Receptors;Staining method;Stains;Structure;Structure of parenchyma of lung;Structure-Activity Relationship;Surveys;System;Techniques;Testing;Tissues;Trachea;Transducers;Vagus nerve structure;Veterans;improved;neuromechanism;operation;pulmonary function;receptive field;receptor;receptor function;relating to nervous system;response;sensor;sensory integration;sensory mechanism;transmission process


application_id: 8043979
project_number: 1I01BX001014-01
project_title: Regulation of adipose cells by autotaxin / lysophosphatidic acid signaling
administering_ic: VA
award_notice_date: 2010-12-02
opportunity_number: BX-10-001
application_type: 1
activity_code: I01
ic: BX
serial_number: 001014
project_start_date: 2010-10-01
project_end_date: 2014-09-30
study_section: Endocriniology A[ENDA]
contact_pi_person_id: 1932626
contact_pi: SMYTH, SUSAN S.
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2010-10-01
project_abstract:
DESCRIPTION (provided by applicant):   
  
Project Summary/Abstract Obesity and obesity-related cardiovascular complications are an increasing health burden in the United States. An analysis of nearly 2 million veterans receiving care at VA Medical Centers in 2000 indicated that ~68% were overweight and ~37% were obese. Despite extensive investigation viable therapeutic strategies to prevent weight gain or promote weight loss remain largely elusive. Attention has recently focused on the exciting possibility that a therapeutic strategy aimed at promoting energy expenditure by increasing brown adipose tissue (BAT) content or activity in adults could reduce fat deposition and obesity. However such a therapeutic strategy requires a comprehensive understanding of factors that regulate BAT. Lysophosphatidic acid (LPA) is a bioactive lipid present in physiologically relevant levels in plasma and other biologic fluids and is positioned to play a role in human health and disease. The secreted lysophospholipase D autotaxin (autotaxin/lysoLPD) is responsible for production of extracellular biologically active LPA which is in turn inactivated by enxymatic dephosphorylation by lipid phosphate phosphatases (LPP). We provide evidence that treatment of primary cultures of preadipocytes with LPA or autotaxin reduces expression of markers of brown adipocyte differentiation and LPP3 an endogenous regulator of Wnt; whereas a novel autotaxin/lysoPLD inhibitor dramatically promotes brown adipocyte differentiation. In mice overexpression of autotaxin reduces UCP1 expression and promotes diet-induced obesity. We therefore propose the central hypothesis that signaling pathways regulated by autotaxin and LPP3 contribute to diet-induced thermogenesis and regulate the development of diet-induced obesity. We will test our central hypothesis using state-of-the art genetic and pharmacologic approaches in three specific aims. In Aim One we will identify the role of autotaxin/lysoPLD in regulation of brown adipose tissue and diet-induced obesity in animal models. In Aim Two we will establish the molecular mechanism(s) by which LPA and autotaxin exert their effects. In Aim Three we identify the role of LPP3 in development and function of adipose tissue and diet-induced obesity. The aims of this grant provide a vehicle to address a major unresolved issue in the field of lysolipid signaling namely role of autotaxin/lysoPLD LPA and LPP3 in regulation of adiposity. These results will be significant because they are expected to provide innovative targets and provide proof-of-concept for novel inhibitors that may be used for prevention and treatment of obesity in humans.  
  
PUBLIC HEALTH RELEVANCE:   
  
Project Narrative More than 44 million Americans are obese as defined by body mass index (BMI) of >30 kg/m2. Obesity is a significant health care problem among veterans receiving care at VA Medical Centers. A comprehensive analysis of nearly 2 million veterans in 2000 indicated that ~68% were overweight and ~37% were obese. Current strategies to treat and prevent obesity focus primarily on reducing energy consumption through appetite suppression and behavioral modification. Recent attention has focused on an attractive alternative approach namely that of increasing energy expenditure by enhancing the heat-dissipating function of brown adipose tissue in adults. We provide evidence that the secreted lysophospholipase D autotaxin responsible for production of biologically active LPA may regulate the functional status of brown adipose tissue. The goal of the current study is to identify the role of the autotaxin/LPA signaling nexus in the regulation of adipose tissue and diet-induced obesity. Our results may suggest innovative targets to prevent and treat obesity in humans.

project_terms:
Address;Adipocytes;Adipose tissue;Adult;Affect;Animal Model;Attention;Attenuated;Body Weight decreased;Brown Fat;Cardiovascular system;Caring;Cell surface;Data;Deposition;Development;Diet;Disease;Down-Regulation;Energy Metabolism;Exhibits;Family;Fatty acid glycerol esters;G-Protein-Coupled Receptors;Gap Junctions;Generations;Genetic;Goals;Grant;Health;Human;In Vitro;Integral Membrane Protein;Investigation;Knowledge;Lipids;Liquid substance;Lysophospholipids;Medical center;Molecular;Mus;Names;Obesity;Organism;Overweight;Plasma;Play;Positioning Attribute;Prevention;Production;Protein Dephosphorylation;Reagent;Recombinants;Regulation;Reporting;Research;Role;Signal Pathway;Signal Transduction;System;Techniques;Testing;Therapeutic;Thermogenesis;Tissue Differentiation;United States;Veterans;Weight Gain;Work;abstracting;adipocyte biology;adipocyte differentiation;alkylglycerophosphoethanolamine phosphodiesterase;extracellular;improved;in vivo;inhibitor/antagonist;innovation;lipid phosphate phosphatase;lysophosphatidic acid;member;novel;obesity prevention;obesity treatment;overexpression;precursor cell;prevent;tool;transdifferentiation


application_id: 8042858
project_number: 1I01CX000240-01A1
project_title: Mechanisms of hepcidin effects in the anemia of chronic disease
administering_ic: VA
award_notice_date: 2010-11-30
opportunity_number: CX-10-001
application_type: 1
activity_code: I01
ic: CX
serial_number: 000240
suffix: A1
project_start_date: 2010-10-01
project_end_date: 2014-09-30
study_section: Hematology[HEMA]
contact_pi_person_id: 1876507
contact_pi: MEANS, ROBERT T
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
project_abstract:
DESCRIPTION (provided by applicant):   
  
The anemia of chronic disease (ACD) is one of the most common clinical syndromes encountered in the practice of medicine. This disorder typically manifests itself as a hypoproliferative anemia accompanied by a low serum iron concentration despite adequate reticuloendothelial iron stores. ACD has generally been considered to result from a combination of pathologic processes which can be linked to the cytokine mediators of inflammation. However the diagnostic importance of altered iron metabolism in ACD has led many investigators to consider that this feature denotes the dominant pathophysiologic mechanism of this syndrome. Hepcidin is an antibacterial protein which is produced in the liver circulates in the blood and is excreted in the urine. It is a type II acute phase protein the expression of which is induced by interleukin (IL)-6 and down-regulated by tumor necrosis factor (TNF). The specific iron regulatory functions of hepcidin as well as the anemia syndromes observed in patients and transgenic animals with abnormalities of hepcidin gene expression (discussed below) have led many to consider that it is the key to unlocking "the mysteries of ACD ".  Three major processes are involved in the pathogenesis of ACD. A slight shortening of red cell survival creates a demand for a small increase in red cell production by the bone marrow. The marrow cannot respond adequately to this demand due to impaired erythropoiesis and impaired mobilization of reticuloendothelial system iron stores. A potential role for hepcidin in the iron anomalies of ACD is strongly supported by available data. However iron changes are not the sole abnormality characteristic of ACD. If hepcidin is the major factor responsible for ACD then it should also contribute to the impaired erythropoiesis observed in this syndrome.  The hypothesis that hepcidin contributes to impaired erythropoiesis in ACD will be addressed through the following specific aims: 1. Determination of hepcidin effects on erythroid colony formation and investigation of the mechanisms involved. This will include A. whether the effects of hepcidin on erythropoiesis are direct-acting or require an accessory cell; B. the role of Epo concentration in hepcidin effects on erythropoiesis; C. the role of apoptosis/anti-proliferation in inhibitory effects of hepcidin on erythropoiesis; D. the effects of cytokines on the hepcidin/erythropoiesis relationship; E. the contributions of iron to hepcidin effects on CFU-E colony formation; F. the role of ferroportin (FPN) and variant FPN in hepcidin effects on CFU-E; and G. effects of Epo and inflammatory cytokines on FPN variant expression in erythroid cells. 2. Determination of the interactions of hepcidin and cytokines in bone marrow macrophage iron transport. Hepcidin inhibits egress of iron from bone marrow macrophages by downregulation of the iron export protein FPN. The ability of inflammatory cytokines to enhance or diminish this effect will be determined as well as any effects on the expression of variant FPN.3. Determination of hepcidin effects on induced Epo production in vitro and delineation of involved mechanisms. This will include A. determination and quantification of the effects of hepcidin on the induction of Epo protein in hepatic and renal models and of the time-course of these effects; and B. determination of the roles of inflammatory cytokines in modulating these effects; C. the role of iron availability in these effects; and D. determination of the effects of hepcidin on Epo transcription and the mechanism involved in these effects. 4. Effects of rhEpo on hepcidin production in vitro. In this Aim we will determine A. the effects of Epo on hepcidin production by HepG2 as well as B. the extent to which these effects are modulated by TNF IL-1 and IL-6. Relevance to VA mission: The proposed studies can potentially lead to an enhanced understanding of the mechanisms of ACD and lead to more effective therapy.  
  
PUBLIC HEALTH RELEVANCE:   
  
Anemia is a major contributor to morbidity and mortality in chronically ill patients and the anemia of chronic disease (ACD) is the major etiology of this anemia. This disorder typically manifests itself as a hypoproliferative anemia accompanied by a low serum iron concentration despite adequate reticuloendothelial iron stores. ACD is highly prevalent in the VA population. In ACD a slight shortening of red cell survival creates a demand for a small increase in red cell production by the bone marrow. The marrow cannot respond adequately to this demand due to impaired erythropoiesis and impaired mobilization of reticuloendothelial system iron stores. A potential role for hepcidin in the iron anomalies of ACD is strongly supported by available data. If hepcidin is the major factor responsible for ACD then it should also contribute to the impaired erythropoiesis observed in this syndrome. The proposed studies can potentially lead to an enhanced understanding of the mechanisms of ACD and therefore to more effective therapy.

project_terms:
Acute-Phase Proteins;Address;Anemia;Anemia due to Chronic Disorder;Anti-Bacterial Agents;Antigen-Presenting Cells;Apoptosis;Blood;Bone Marrow;CFU-E;Cell Survival;Characteristics;Chronically Ill;Clinical;Data;Diagnostic;Disease;Down-Regulation;Erythrocytes;Erythroid;Erythroid Cells;Erythropoiesis;Etiology;Gene Expression;Genetic Transcription;Hepatic;In Vitro;Inflammation Mediators;Inflammatory;Interleukin-1;Interleukin-6;Investigation;Iron;Kidney;Lead;Link;Liver;Marrow;Medicine;Mission;Modeling;Morbidity - disease rate;Mortality Vital Statistics;Pathogenesis;Pathologic Processes;Patients;Population;Process;Production;Protein Export Pathway;Proteins;Research Personnel;Reticuloendothelial System;Role;Serum;Syndrome;Time;Transgenic Animals;Tumor Necrosis Factor-alpha;Urine;Variant;cytokine;effective therapy;hepcidin;iron metabolism;macrophage;metal transporting protein 1;protein expression


application_id: 8003690
project_number: 1I01RX000338-01A1
project_title: TBI epileptogenesis: pathologic hippocampal L-glut synaptic plasticity
administering_ic: VA
award_notice_date: 2010-10-26
opportunity_number: RX-10-005
application_type: 1
activity_code: I01
ic: RX
serial_number: 000338
suffix: A1
project_start_date: 2010-10-01
project_end_date: 2014-09-30
study_section: Brain Health and Injury[RRD1]
contact_pi_person_id: 9405862
contact_pi: SLEVIN, JOHN T.
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
project_abstract:
DESCRIPTION (provided by applicant):   
  
Project Summary The molecular mechanism that underlies evolution to post-traumatic epilepsy (PTE) in individuals with traumatic brain injury (TBI) is unknown. It has been suggested that the development of PTE following TBI is the combined result of kindling epileptogenesis during the early stages of recovery and triggering by residual irritation from damaged neural tissue. Glutamate the major excitatory neurotransmitter (NT) in the central nervous system must be tightly regulated to allow for proper neuronal signaling without creating excessive excitation. TBI produces an increase in glutamate release into the extracellular space and a concurrent decrease in the ability of excitatory amino acid transporters to reuptake glutamate that may lead to its accumulation propagating secondary damage cascades and producing aberrant neuronal signaling. We suggest that the enhanced hippocampal glutamate levels seen post-TBI are in part due to changes of the neurosecretory machinery and SNARE regulators and/or re-sequestration process. Left unchecked these changes become drivers of post-TBI epileptogenesis leading to PTE. The hypothesis that pathologic adaptation of inherent hippocampal glutamatergic synaptic plasticity promotes post-TBI epileptogenesis will be tested through three specific aims in animals (and appropriate controls) that have been administered a lateral fluid percussion injury (LFPI) and then monitored by telemetry until electroencephalographic (EEG) evidence of epileptiform activity: (1) Measure of tonic glutamate levels and evoked glutamateamate release in the hippocampal formation of epileptic and non-epileptic animals after moderate LFPI through use of a novel microarray electrode system. In tissue from hippocampal formation of these same animals we will use Western blotting techniques to quantify (2) components of the neurotransmitter release process and (3) certain glutamate transporters. Investigation of this putative relationship between aberrant glutamate neuronal exocytosis/resequestration and PTE will provide insight into potential mechanisms by which glutamate neurotransmission is regulated and into the molecular events of epileptogenesis as well as provide potential foci for therapeutic intervention. The long-term goal of this project is to restore to normal aberrant mechanisms associated with TBI-associated epileptogenesis. There are indications from animal studies that several newer FDA-approved and actively used antiepileptic drugs e.g. levetiracetam and lacosamide may also act as antiepileptogenic agents through effect on glutamate neurotransmission. Such agents would be appropriate compounds for further mechanistic evaluation in animals and clinical studies in PTE patients if the studies proposed here suggest that chronic dysregulation of glutamatergic release is associated with epileptogenesis. Pretransfection with a recently developed adeno-associated virus for the GLT-1 transporter to enhance glutamate reuptake currently under investigation by one of us may provide yet another novel approach to target a specific mechanism involved in glutamate regulation should these studies confirm a glutamate transporter abnormality is associated with development of PTE.  
  
PUBLIC HEALTH RELEVANCE:   
  
In addition to affecting civilian populations traumatic brain injury (TBI) has been labeled the signature injury of the current combat in Iraq and Afghanistan. TBI is a leading cause of morbidity and mortality in the United States and is a risk factor for post-traumatic epilepsy (PTE). Examining TBI-induced dysregulation of glutamate neurotransmitter release and reuptake the specific aims of this proposal may reveal important mechanisms identify potential therapeutic targets and pinpoint the therapeutic window to eliminate or reduce the probability of developing PTE. Recognition of molecular targets for new therapies that would extinguish TBI- induced epileptogenesis would alleviate the human suffering of civilians soldiers and veterans as they return to civilian life as well as diminish the concomitant long-term economic costs of epilepsy to the US health care system the military and the Veterans Administration.


application_id: 7931267
project_number: 1I01BX000387-01A2
project_title: Mood-Stabilizing Medications and the Inositol Signaling System
administering_ic: VA
award_notice_date: 2010-11-26
opportunity_number: BX-09-008
application_type: 1
activity_code: I01
ic: BX
serial_number: 000387
suffix: A2
project_start_date: 2010-04-01
project_end_date: 2014-03-31
study_section: Neurobiology A[NURA]
contact_pi_person_id: 7118863
contact_pi: PARTHASARATHY, RANGA N
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
project_abstract:
DESCRIPTION (provided by applicant):   
  
Project Summary/Abstract Bipolar disorder (BD) is a devastating condition affecting many Veterans. Lithium and valproate are the drugs of choice for the manic phase of BD. While therapeutically effective their mechanisms of action are not fully understood and undesirable side effects are common. Lithium inhibits brain inositol monophosphatase (IMPase) thereby depleting the available pool of free inositol and resulting in the correction of the manic phase of BD. Inositol synthase (IPS) is a key enzyme that regulates inositol levels by generating the precursor inositol phosphate (IP). Our preliminary data demonstrate that rat brain IPS is up-regulated by chronic lithium treatment which would have the same net effect as inositol depletion relative to inositol phosphate. We hypothesize that lithium and other mood stabilizers exert their therapeutic effects by up-regulating IPS expression increasing the available inositol phosphate pool. This implies that the pathophysiological mechanism underlying BD is a disturbance in maintaining a proper balance in the ratio of free inositol to inositol phosphate pools. Our long-term objective is to define how mood stabilizers work and to find common therapeutic targets in the pathophysiology of BD so new medications can be designed. Our specific aims are: 1) Determine the changes in abundance and activity of the newly-identified isoforms of IPS in brain regions after chronic lithium and valproate; 2) Determine the effects of lithium and valproate on IPS binding to clathrin and receptor-mediated endocytosis; 3) Establish the link between inositol metabolism and behavior in IPS-deficient mice; and 4) Identify drug-targetable convergent pathways for lithium valproate and lamotirgine responsive genes. To accomplish Aim 1 IPS and its isoforms will be quantified in drug-treated rat brain regions by enzyme activity assay Western blot and Q-PCR methods. To accomplish Aim 2 brain IPS and its complexes will be analyzed by Western and mass spectral analyses. Effects on receptor-mediated endocytosis will be monitored by following transferrin receptor trafficking and fluorescence resonance energy transfer (FRET) analysis. For Aim 3 lithium and valproate treated and untreated IPS knock-out mice will be subjected to standardized behavioral tests and their brains assayed for key enzymes in inositol metabolism. Aim 4 will be addressed by microarray and "pathway" analyses to map and compare the chronic lithium valproate and lamotrigine transcriptomes in human neuronal cell lines and in rat primary cortical neuronal cell cultures. Defining the mechanisms of lithium action on inositol metabolism in the brain from the perspective of IPS function may establish a direct connection between inositol regulation and behavior. Thus advancing our understanding of the pathophysiology of BD and disclosing new therapeutic targets for treating BD.  
  
PUBLIC HEALTH RELEVANCE:   
  
PROJECT NARRATIVE (Potential Relevance to Veterans' Health) Bipolar Disorder (BD) is a major clinical problem for Veterans and there are many Iraq War Veterans treated by VA Psychiatry Service for lithium-resistant BD. To better manage these patients we must be able to identify common targets of mood-stabilizers so that future medications can be designed that have greater efficacy and fewer side effects. One way lithium can correct BD is by modulating brain inositol signal pathways. The brain maintains inositol (a sugar) homeostasis through three routes. Inositol phosphate synthase (IPS) is the only biosynthetic means of generating inositol from glucose. IPS is a new target linked to the genesis of BD. Stimulators of brain IPS enzyme which will then cause the generation of inositol inside the human brain can be considered as an important new class of mood stabilizing medications. The results from these studies will provide a highly positive impact on the health of Veterans with BD.

project_terms:
Address;Adverse effects;Affect;Behavior;Behavior Disorders;Behavioral;Binding (Molecular Function);Biochemical;Biological Assay;Biological Process;Bipolar Disorder;Brain;Brain region;Cell Culture Techniques;Cell Line;Cell physiology;Cells;Cerebellum;Chronic;Clathrin;Clinical;Complex;Corpus striatum structure;Data;Drug Delivery Systems;Drug resistance;Endocytosis;Enzymatic Biochemistry;Enzymes;Equilibrium;Fluorescence Resonance Energy Transfer;Foundations;Functional disorder;Future;Gene Expression Profile;Generations;Genes;Global Change;Glucose;Goals;Health;Hippocampus (Brain);Homeostasis;Human;In Vitro;Individual;Inositol;Inositol Metabolism Pathway;Inositol Phosphates;Iraq;Knockout Mice;Link;Lithium;Manic;Manic Disorder;Maps;Measurement;Measures;Mediating;Mental Depression;Messenger RNA;Methods;Modeling;Monitor;Mood stabilizers;Moods;Mus;Mutation;Neurons;Pathway interactions;Patients;Pharmaceutical Preparations;Phospholipids;Phosphorylation;Predisposition;Prefrontal Cortex;Protein Isoforms;Proteins;Psychiatry;RNA;Rattus;Regulation;Relative (related person);Resistance;Route;Services;Signal Pathway;Signal Transduction;System;Testing;Therapeutic;Therapeutic Effect;Time;Transferrin Receptor;Treatment Efficacy;Veterans;War;Western Blotting;Work;abstracting;behavior test;brain pathway;chromatin remodeling;design;diagnostic accuracy;enzyme activity;feeding;gene interaction;improved;lamotrigine;membrane synthesis;mouse model;myo-Inositol-1-Phosphate Synthase;myo-inositol-1 (or 4)-monophosphatase;new therapeutic target;novel;protein protein interaction;receptor mediated endocytosis;research study;response;sugar;therapeutic target;trafficking;valproate


application_id: 7906849
project_number: 7I01BX000216-02
project_title: Nicotine-induced fibronectin expression in lung injury and repair
administering_ic: VA
award_notice_date: 2010-12-16
opportunity_number: BX-09-001
application_type: 7
activity_code: I01
ic: BX
serial_number: 000216
project_start_date: 2009-10-01
project_end_date: 2014-03-31
study_section: Respiration[PULM]
contact_pi_person_id: 1876531
contact_pi: ROMAN, JESSE
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2010-10-01
project_abstract:
DESCRIPTION (provided by applicant):  ABSTRACT: Nicotinic acetylcholine receptors (nAChRs) are pentameric channels expressed by neuronal and non-neuronal cells that are capable of signal transduction. Although much of what is known about the function of nAChRs relates to the nervous system information about their roles in non-neuronal systems is rapidly emerging. nAChRs are expressed in lung and interesting work performed in non-human primates suggests that nAChR activation by nicotine leads to pulmonary structural/functional abnormalities. We explored the mechanisms by which nAChR activation leads to matrix expression and found that this effect is mediated through a specific subgroup of nAChRs termed 17 nAChRs. Upon activation of these receptors in lung fibroblasts we observed induction of the MEK-1/Erk pathway and phosphorylation of the transcription factor CREB. In turn CREB stimulated the expression of fibronectin a matrix glycoprotein implicated in lung injury and repair. These events were blocked by 17 nAChR antagonists and were not present in 17 nAChR KO primary lung fibroblasts. In addition we found that nAChR activation stimulates lung fibroblast proliferation through MEK-1/Erk-dependent pathways. Similar observations were made in mice chronically stimulated with nicotine but not in developing and adult 17 nAChR KO animals. More recently we documented an increase in the proliferation and expression of fibronectin in fibroblasts exposed to cigarette smoke extract and observed an increase in fibronectin protein deposition in the lungs of mice exposed to tobacco smoke confirming the relevance of this process to tobacco-related disease. We also found that lung repair mechanisms appeared to be impaired after bleomycin-induced lung injury in the setting of chronic exposure to nicotine. These data unveil a novel role for non-neuronal nAChRs in control of lung tissue remodeling. Based on the above we hypothesize that 17 nAChR activation in lung promotes tissue remodeling by stimulating the proliferation of fibroblasts and their overexpression of matrix components like fibronectin through MEK-1/Erk-dependent signals. At baseline this leads to subtle alterations in matrix composition without causing overt consequences such as fibrosis. In contrast in the setting of lung injury this leads to exaggerated tissue remodeling that promotes inefficient repair and fibrosis. This will be tested in specific aims designed to: 1) Examine the role of 17 nAChRs in the proliferation myofibroblast transdifferentiation and matrix expression of lung fibroblasts and the role of MEK-1/Erk in mediating these events in wildtype vs. 17 nAChR KO lung fibroblasts 2) Examine the role of 17 nAChRs in lung injury and repair using wildtype and 17 nAChR KO mice in two models of lung injury (bleomycin model and the model of heterologous tracheal transplantation) and 3) Examine the effect of persistent 17 nAChR activation in lung repair after injury.  PUBLIC HEALTH RELEVANCE:  Project Narrative This project will explore the cellular mechanisms involved in tissue remodeling of the lung after injury. Tissue remodeling is a process characterized by alterations in the expression deposition and degradation of connective tissue components (also called extracellular matrices). This is important to active combatants but also to veterans in view that many pulmonary disorders typically seen in this population (e.g. COPD lung fibrosis) that are associated with alterations in tissue remodeling. Interestingly tobacco and its components have been implicated in many of these disorders and therefore it is thought that they promote alterations in tissue remodeling. But the mechanisms responsible for these events are unknown. This project will explore the mechanisms involved in these processes by focusing on nicotinic receptors and how activation of these receptors promotes alterations in the relative composition of the lung extracellular matrix.

project_terms:
Adult;Affect;Animals;Bleomycin;Bronchoalveolar Lavage Fluid;Bronchopulmonary Dysplasia;CREB1 gene;Cells;Childhood;Chronic;Chronic Obstructive Airway Disease;Clinical Research;Collagen;Connective Tissue;Data;Deposition;Disease;Event;Exposure to;Extracellular Matrix;Fibroblasts;Fibronectins;Fibrosis;Genes;Genetic Transcription;Glycoproteins;Hamman-Rich syndrome;Harvest;Health;In Vitro;Inbred C57BL Mice;Injury;Ion Channel;Knock-out;Knockout Mice;Link;Lung;Lung diseases;Mediating;Messenger RNA;Modeling;Morphogenesis;Mus;Mutation;Myofibroblast;Neuroglia;Neurons;Nicotine;Nicotinic Receptors;Pathway interactions;Phosphorylation;Play;Population;Process;Production;Promotor (Genetics);Protein Isoforms;Proteins;Receptor Activation;Relative (related person);Reporting;Role;Signal Transduction;Site;Structure of parenchyma of lung;Subgroup;System;Testing;Time;Tissues;Tobacco;Tobacco smoke;Tobacco use;Transplantation;Veterans;Work;alpha-bungarotoxin receptor;base;cigarette smoke;design;drinking water;feeding;human MAP2K1 protein;in vivo;injured;injury and repair;interest;lung development;lung injury;mTOR Signaling Pathway;nonhuman primate;novel;outcome forecast;overexpression;prenatal;protein expression;receptor;relating to nervous system;repaired;response;transcription factor;transdifferentiation

public_health_relevance:
Project Narrative This project will explore the cellular mechanisms involved in tissue remodeling of the lung after injury. Tissue remodeling is a process characterized by alterations in the expression deposition and degradation of connective tissue components (also called extracellular matrices). This is important to active combatants but also to veterans in view that many pulmonary disorders typically seen in this population (e.g. COPD lung fibrosis) that are associated with alterations in tissue remodeling. Interestingly tobacco and its components have been implicated in many of these disorders and therefore it is thought that they promote alterations in tissue remodeling. But the mechanisms responsible for these events are unknown. This project will explore the mechanisms involved in these processes by focusing on nicotinic receptors and how activation of these receptors promotes alterations in the relative composition of the lung extracellular matrix.


application_id: 7905753
project_number: 5I01BX000325-02
project_title: Studies on the role of IL-23p19/IL-17 cascade in the pathogenesis of acute GVHD
administering_ic: VA
award_notice_date: 2010-11-29
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000325
project_start_date: 2009-10-01
project_end_date: 2012-12-31
study_section: Immunology and Dermatology A[IMMA]
contact_pi_person_id: 8781506
contact_pi: THOMPSON, JOHN SECORD
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2010-10-01
project_abstract:
DESCRIPTION (provided by applicant):  Abstract It is firmly established that aGVHD is initiated by T-cell recognition of antigenic foreignness and ex vivo depletion of T-cells has been the most effective method to reduce its incidence and severity. The proposed studies are to test 2 hyptheses: 1) The IL-23p19/IL-17 cytokine axis plays a major role in the inflammatory reactions characteristic of aGVHD by regulating the expression of key cytokine genes that may promote or inhibit aGVHD. 2) The IL -23p19/1L-17 cytokine axis interacts closely but not completely with the TNF1 IL-12 and IL-6 cascade to amplify the pathology of aGVHD. Three objectives are designed to address these They are derived from our observation that BALB/c mice transplanted with bone marrow (BM) plus T- containing spleens (SCs) from mutant mice deficient in the p19 dimer of the IL-23 p19/p40 heterodimer develop aGVHD less frequently and less severely than BALB/c transplanted with normal wild type C57BL/6 BM + SCs. IL-17 mRNA and serum cytokine produced by CD8 Th17 cells are elevated in BALB/c mice transplanted with both WT and p19 deficient cells but to significantly higher levels in the WT animals. Induction of Th17 producing IL-17 in the p19-/- mice was shown to be derived from TGF2 IL-6 and IL23p19 induced by total body irradiation of the host BALB/c animals. The first objective is to determine the effect on the induction of aGVHD by isolated T and other cells in the donor inoculum. The second objective is to determine the role of the products of Th17 cells in the pathogenesis of aGVHD. The third objective is to determine the relative roles of the IL-23/IL17-21-22 cascade versus the TNF1-IL-12-IL-6 cascade in the pathology of aGVHD. Use of target mutant animals deficient in IL-23p19 RORgamma t (critical for Th17 cell development) and TNF1 provide unique models in which to dissect the components of aGVHD pathology.  PUBLIC HEALTH RELEVANCE:  Relevance to VA Mission: Every day large numbers of unfortunate veterans present with new cancers. In addition to the leukemias myelodysplasia and multiple myeloma other cancers could be amenable to stem cell transplantation. Admittedly there are some cancers that probably would not benefit from this therapy but the ability to achieve a revitalized immune system could be an important addition to their care. However as stated above the current risk of developing transplant related toxicities GVHD and overwhelming infection are too great to offer stem cell transplantation to patents with a wider variety of cancers. The proposed studies are to investigate a new approach to minimize GVHD

project_terms:
Acute;Acute Graft Versus Host Disease;Address;Allogenic;Animals;Bone Marrow;CD8B1 gene;Caring;Cells;Characteristics;Chronic Graft Versus Host Disease;Cytokine Gene;Development;Disease;Dysmyelopoietic Syndromes;Graft-vs-Host Disease;Health;IL-23 p19;IL17 gene;Immune system;Inbred BALB C Mice;Incidence;Infection;Inflammatory;Interleukin-12;Interleukin-17;Interleukin-6;Legal patent;Malignant Neoplasms;Messenger RNA;Methods;Mission;Modeling;Multiple Myeloma;Mus;Mutant Strains Mice;Pathogenesis;Pathology;Play;Reaction;Relative (related person);Risk;Role;Serum;Severities;Spleen;Stem cell transplant;Survivors;T-Cell Depletion;T-Lymphocyte;Testing;Toxic effect;Transplantation;Veterans;Whole-Body Irradiation;abstracting;cytokine;design;dimer;experience;interleukin-23;leukemia;mutant;novel strategies

public_health_relevance:
Relevance to VA Mission: Every day large numbers of unfortunate veterans present with new cancers. In addition to the leukemias myelodysplasia and multiple myeloma other cancers could be amenable to stem cell transplantation. Admittedly there are some cancers that probably would not benefit from this therapy but the ability to achieve a revitalized immune system could be an important addition to their care. However as stated above the current risk of developing transplant related toxicities GVHD and overwhelming infection are too great to offer stem cell transplantation to patents with a wider variety of cancers. The proposed studies are to investigate a new approach to minimize GVHD


application_id: 7904139
project_number: 5I01BX000672-02
project_title: Class B Scavenger Receptors and Atherosclerosis
administering_ic: VA
award_notice_date: 2010-11-29
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000672
project_start_date: 2009-10-01
project_end_date: 2013-09-30
study_section: Cardiovascular Studies A[CARA]
contact_pi_person_id: 7875298
contact_pi: VAN DER WESTHUYZEN, DENEYS REM
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2010-10-01
project_abstract:
DESCRIPTION (provided by applicant):   
  
The roles of the Class B scavenger receptors SR-BI and CD36 in the development of atherosclerotic disease a major cause of death remain poorly understood. This is due in part to the varied and still little understood functions of these two pattern recognition receptors in host defense and tissue homeostasis. The role of CD36 in the initiation of macrophage foam cell formation is controversial although CD36 has the ability to take up modified lipoproteins such as oxidized LDL. Other potentially important functions of CD36 include its role in the innate immune system and its role in inflammatory signaling. Atherosclerosis is a chronic inflammatory disease and we propose that CD36 plays a more complex role in atherosclerotic lesion formation. The structurally closely related receptor SR-BI plays a key role in reverse cholesterol transport to the liver and has been shown in several mouse models to be anti-atherogenic. Recent findings have also indicated additional extrahepatic roles of SR-BI in atherogenesis including effects in macrophages. The functional basis for these effects is not known although we have shown that SR-BI-null mice have a markedly enhanced inflammatory cytokine response and higher lethality in response to lipopolysaccharide (LPS). Furthermore mice deficient in both SR-BI and CD36 unexpectedly exhibit much greater atherosclerotic lesion formation than mice lacking either SR-BI or CD36 only. These findings indicate that SR-BI and CD36 each have athero-protective functions yet to be defined. The protective functions of the two receptors may be overlapping allowing at least partial compensation in mice lacking only one of the two receptors. The central hypothesis of this proposal is that SR-BI and CD36 act in a complementary or synergistic manner to protect against atherosclerosis. We propose that SR-BI and CD36 are key to the innate defense against atherosclerosis a chronic inflammatory disease. The following 4 objectives are proposed to examine these hypotheses: Objective 1 will examine the complementary roles SR-BI and CD36 in atherosclerotic lesion formation in response to high fat diets. This will be accomplished by analyzing the extent and nature of aortic lesions in wild type SR-BI-null CD36-null and SR-BI-nullXCD36-null mice. Objective 2 will investigate the roles of macrophage-expressed Class B scavenger receptors in atherosclerosis. This will be accomplished through the use of bone marrow transplantation. Objective 3 will identify the complementary or synergistic roles of SR-B1 and CD36 in macrophage foam cell formation. This will be accomplished by measuring lipid accumulation in wild type and SR-BI-XCD36- DKO macrophages treated with modified LDLs. Cells will be examined for lipid related protein expression and by gene microarray analysis. Lipid uptake and efflux pathways will be compared. Objective 4 will investigate enhanced inflammatory signaling in CD36/SR-BI DKO macrophages and its effects on macrophage lipid accumulation. The impact of inflammatory signaling on modified LDL uptake in primary macrophages will be evaluated together with the hypothesis that increased plasma membrane cholesterol content in SR-BIxCD36 DKO macrophages stimulates inflammatory signaling pathways. These studies are expected to provide new understanding of the metabolic functions of CD36 and SR-BI as well as their role in atherosclerosis.  
  
PUBLIC HEALTH RELEVANCE:   
  
Cardiovascular disease remains the number one killer in the United States and among veterans. Although widely studied the mechanisms underlying atherosclerosis are not yet clearly understood. Plasma cholesterol and LDL levels are a major risk factor for atherosclerosis but the underlying mechanisms for this are still poorly understood. Further understanding of how LDL cholesterol accumulates in the artery wall and the roles of the various receptors for lipoproteins will potentially allow for the identification of new treatment strategies and new drug development.

project_terms:
Affect;Antiatherogenic;Arterial Fatty Streak;Arteries;Atherosclerosis;Bone Marrow Transplantation;CD36 gene;Cardiovascular Diseases;Cause of Death;Cell membrane;Cells;Cholates;Cholesterol;Chronic;Complex;Development;Diet;Disease;Exhibits;Extrahepatic;Fatty acid glycerol esters;Financial compensation;Foam Cells;Genes;Hepatic;Homeostasis;Host Defense;Immune system;Inflammation;Inflammatory;Inflammatory Response Pathway;Knock-out;Knockout Mice;LDL Cholesterol Lipoproteins;Lesion;Lipids;Lipopolysaccharides;Lipoprotein Receptor;Lipoproteins;Liver;Low-Density Lipoproteins;Measures;Metabolic;Metabolism;Microarray Analysis;Mus;Nature;Pathway interactions;Pattern recognition receptor;Plasma;Play;Regulation;Risk Factors;Role;SR-B proteins;SR-BI receptor;Signal Pathway;Signal Transduction;TLR4 gene;Testing;Tissues;Transplantation;United States;Veterans;atherogenesis;base;drug development;feeding;macrophage;mouse model;oxidized low density lipoprotein;protein expression;public health relevance;receptor;research study;response;reverse cholesterol transport;scavenger receptor;treatment strategy;uptake

public_health_relevance:
Cardiovascular disease remains the number one killer in the United States and among veterans. Although widely studied the mechanisms underlying atherosclerosis are not yet clearly understood. Plasma cholesterol and LDL levels are a major risk factor for atherosclerosis but the underlying mechanisms for this are still poorly understood. Further understanding of how LDL cholesterol accumulates in the artery wall and the roles of the various receptors for lipoproteins will potentially allow for the identification of new treatment strategies and new drug development.


application_id: 7903353
project_number: 5I01BX000610-02
project_title: Analysis of PTH and Dopamine Receptor Signaling in Proximal Tubules
administering_ic: VA
award_notice_date: 2010-11-29
opportunity_number: BX-09-001
application_type: 5
activity_code: I01
ic: BX
serial_number: 000610
project_start_date: 2009-10-01
project_end_date: 2013-09-30
study_section: Nephrology[NEPH]
contact_pi_person_id: 2262379
contact_pi: LEDERER, ELEANOR D
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2010-10-01
project_abstract:
DESCRIPTION (provided by applicant):  Project Summary/Abstract The goal of this project is to determine how the structural protein NHERF-1 sodium-hydrogen exchanger regulatory factor isoform 1 regulates the trafficking of Npt2a the type IIa sodium phosphate cotransporter to the apical membrane of the renal proximal tubule. The expression of Npt2a at the apical membrane is a critical regulatory step because the level of expression and function of Npt2a is the primary regulator of total body phosphate homeostasis. VA-funded research from this laboratory had revealed that the absence of NHERF-1 in a model of proximal renal tubule OK (opossum kidney) cells resulted in a marked decrease in Npt2a expression. The decrease in expression was due to two factors 1) decreased transcription of Npt2a and 2) decreased trafficking of Npt2a to the apical membrane. The mechanisms for the faulty trafficking of Npt2a have not been determined and are the subject of this proposal. Npt2a translated in the NHERF-deficient cells (OK-H) lacked a critical post-translational modification glycosylation. These proteins instead of trafficking to apical membrane accumulated in a perinuclear location. Inefficient apical membrane localization had previously been described in a mouse lacking expression of NHERF-1. Expression of a NHERF-1 construct that lacked a normal PDZ-2 domain did not allow trafficking of Npt2a to the apical membrane. NHERF-1 is a multiple PDZ domain protein which interacts with multiple signaling molecules and transporter proteins. A role for the PDZ-2 domain in Npt2a regulation had not been observed previously. Finally inhibition of SNARE (SNAP Receptor) protein interaction using a competing peptide introduced into OK cells blocked insertion of Npt2a into the apical membrane demonstrating that Npt2a is transported to the apical membrane via a vesicular transport mechanism. The preliminary data suggested the hypothesis that NHERF-1 is essential for the forward trafficking of Npt2a from site of synthesis to the apical membrane. First the role of the post-translational modifications glycosylation and phosphorylation on Npt2a apical membrane trafficking will be examined by determining the cellular localization of transfected Npt2a constructs lacking the motifs required for these post translational modifications in a cell culture model. The glycosylation phosphorylation and ubiquitination states of Npt2a will be compared in NHERF-replete and NHERF-deficient OK cells and in wild type and NHERF-1 knock out mouse proximal tubule cells. The effect of inhibitors of glycosylation on the intracellular localization of Npt2a will be determined. Second the intracellular site or sites of interaction between NHERF-1 and Npt2a will be analyzed. Forward trafficking of Npt2a labeled by a GFP tag and by S35 methionine in OK cells and OK-H cells will be slowed by culture at 20 C. Analysis of Npt2a localization by confocal imaging density centrifugation and immunoelectronmicroscopy will be performed at sequential time points until the proteins are detected as biotinylated forms indicating appropriate insertion into the apical membrane. Trafficking of Npt2a in the two cell culture models will be compared. Proteins associated with Npt2a at each time point will be determined by immunoprecipitation and proteomic analysis. To define which steps in the forward trafficking are NHERF- dependent Npt2a trafficking will be compared in NHERF-replete and NHERF-deficient cells under conditions of traffic arrest: ezrin deficiency inhibition of SNARE interaction and CLC-5 (intracellular chloride channel CLC family isoform 5) deficiency. Third the sites on Npt2a and NHERF-1 critical for the NHERF-1 effect on Npt2a trafficking will be determined by mutational analysis of both proteins followed by both in vitro analysis of protein interaction and assessment of intracellular interaction by FRET methodology. These studies will define where NHERF-1 acts in Npt2a forward trafficking define the sites on both proteins responsible for their interaction and yield mechanistic insights for this unique functional process.  PUBLIC HEALTH RELEVANCE:  Project Narrative The goal of this project is to determine how the protein NHERF-1 sodium-hydrogen exchanger regulatory factor isoform 1 regulates the trafficking of Npt2a the type IIa sodium phosphate cotransporter to the apical membrane of the renal proximal tubule. The kidney regulates the total body content of phosphate through altering the expression of Npt2a at the apical membrane. Phosphate wasting can cause kidney stones and metabolic bone disease while phosphate retention is a risk factor for cardiovascular events common diseases in Veterans of all ages. Cardiovascular disease is the major cause of morbidity and mortality in the Veteran population. Kidney stones tend to affect individuals during their most productive years resulting in over $2 billion dollars of medical expenses in this country. Metabolic bone disease contributes substantially to the occurrence of hip fracture a disabling event with up to a 50% one year mortality. Thus understanding how the kidney regulates phosphate homeostasis has significant relevance to the health of Veterans of all ages.

project_terms:
Address;Affect;Age;Apical;CLC Gene;Cardiovascular Diseases;Carrier Proteins;Cell Culture Techniques;Cell Line;Cells;Centrifugation;Chloride Channels;Country;Data;Didelphidae;Disease;Dopamine Receptor;Event;Exhibits;Family;Fluorescence Resonance Energy Transfer;Funding;Genetic Transcription;Goals;Health;Hip Fractures;Homeostasis;Image;Immunoprecipitation;In Vitro;Individual;Kidney;Kidney Calculi;Knockout Mice;Label;Laboratories;Laboratory Research;Location;Medical;Membrane Microdomains;Membrane Protein Traffic;Membrane Transport Proteins;Metabolic Bone Diseases;Methionine;Methodology;Modeling;Molecular;Morbidity - disease rate;Mortality Vital Statistics;Mus;Mutation;Pathway interactions;Peptides;Phosphorylation;Physiologic pulse;Play;Population;Post-Translational Protein Processing;Process;Protein Analysis;Protein Isoforms;Proteins;Proteomics;Proximal Kidney Tubules;Receptor Signaling;Regulation;Renal function;Research;Research Personnel;Role;SNAP receptor;Signaling Molecule;Site;Staging;Structural Protein;Tertiary Protein Structure;Time;TimeLine;Transfection;Translating;Transport Vesicles;Tubular formation;Ubiquitination;Vesicle;Veterans;Work;abstracting;apical membrane;base;cardiovascular risk factor;caveolin 1;cold temperature;density;design;ezrin;glycosylation;in vitro Assay;inhibitor/antagonist;inorganic phosphate;insight;kidney cell;mutant;protein expression;receptor;research study;sodium-hydrogen exchanger regulatory factor;sodium-phosphate cotransporter proteins;solute;syntaxin binding protein 1;trafficking;wasting

public_health_relevance:
Project Narrative  The goal of this project is to determine how the protein NHERF-1 sodium-hydrogen exchanger regulatory factor isoform 1 regulates the trafficking of Npt2a the type IIa sodium phosphate cotransporter to the apical membrane of the renal proximal tubule. The kidney regulates the total body content of phosphate through altering the expression of Npt2a at the apical membrane. Phosphate wasting can cause kidney stones and metabolic bone disease while phosphate retention is a risk factor for cardiovascular events common diseases in Veterans of all ages. Cardiovascular disease is the major cause of morbidity and mortality in the Veteran population. Kidney stones tend to affect individuals during their most productive years resulting in over $2 billion dollars of medical expenses in this country. Metabolic bone disease contributes substantially to the occurrence of hip fracture a disabling event with up to a 50% one year mortality. Thus understanding how the kidney regulates phosphate homeostasis has significant relevance to the health of Veterans of all ages.


application_id: 7797740
project_number: 1I01BX000325-01A1
project_title: Studies on the role of IL-23p19/IL-17 cascade in the pathogenesis of acute GVHD
administering_ic: VA
award_notice_date: 2009-07-31
opportunity_number: BX-09-001
application_type: 1
activity_code: I01
ic: BX
serial_number: 000325
suffix: A1
project_start_date: 2009-10-01
project_end_date: 2012-09-30
study_section: Immunology and Dermatology A[IMMA]
contact_pi_person_id: 8781506
contact_pi: THOMPSON, JOHN SECORD
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
project_abstract:
DESCRIPTION (provided by applicant):   
  
Abstract It is firmly established that aGVHD is initiated by T-cell recognition of antigenic foreignness and ex vivo depletion of T-cells has been the most effective method to reduce its incidence and severity. The proposed studies are to test 2 hyptheses: 1) The IL-23p19/IL-17 cytokine axis plays a major role in the inflammatory reactions characteristic of aGVHD by regulating the expression of key cytokine genes that may promote or inhibit aGVHD. 2) The IL -23p19/1L-17 cytokine axis interacts closely but not completely with the TNF1 IL-12 and IL-6 cascade to amplify the pathology of aGVHD. Three objectives are designed to address these They are derived from our observation that BALB/c mice transplanted with bone marrow (BM) plus T- containing spleens (SCs) from mutant mice deficient in the p19 dimer of the IL-23 p19/p40 heterodimer develop aGVHD less frequently and less severely than BALB/c transplanted with normal wild type C57BL/6 BM + SCs. IL-17 mRNA and serum cytokine produced by CD8 Th17 cells are elevated in BALB/c mice transplanted with both WT and p19 deficient cells but to significantly higher levels in the WT animals. Induction of Th17 producing IL-17 in the p19-/- mice was shown to be derived from TGF2 IL-6 and IL23p19 induced by total body irradiation of the host BALB/c animals. The first objective is to determine the effect on the induction of aGVHD by isolated T and other cells in the donor inoculum. The second objective is to determine the role of the products of Th17 cells in the pathogenesis of aGVHD. The third objective is to determine the relative roles of the IL-23/IL17-21-22 cascade versus the TNF1-IL-12-IL-6 cascade in the pathology of aGVHD. Use of target mutant animals deficient in IL-23p19 RORgamma t (critical for Th17 cell development) and TNF1 provide unique models in which to dissect the components of aGVHD pathology.  
  
PUBLIC HEALTH RELEVANCE:   
  
Relevance to VA Mission: Every day large numbers of unfortunate veterans present with new cancers. In addition to the leukemias myelodysplasia and multiple myeloma other cancers could be amenable to stem cell transplantation. Admittedly there are some cancers that probably would not benefit from this therapy but the ability to achieve a revitalized immune system could be an important addition to their care. However as stated above the current risk of developing transplant related toxicities GVHD and overwhelming infection are too great to offer stem cell transplantation to patents with a wider variety of cancers. The proposed studies are to investigate a new approach to minimize GVHD

project_terms:
Acute;Acute Graft Versus Host Disease;Address;Allogenic;Animals;Bone Marrow;CD8B1 gene;Caring;Cells;Characteristics;Chronic Graft Versus Host Disease;Cytokine Gene;Development;Disease;Dysmyelopoietic Syndromes;Graft-vs-Host Disease;Health;IL-23 p19;IL17 gene;Immune system;Inbred BALB C Mice;Incidence;Infection;Inflammatory;Interleukin-12;Interleukin-17;Interleukin-6;Legal patent;Malignant Neoplasms;Messenger RNA;Methods;Mission;Modeling;Multiple Myeloma;Mus;Mutant Strains Mice;Pathogenesis;Pathology;Play;Reaction;Relative (related person);Risk;Role;Serum;Severities;Spleen;Stem cell transplant;Survivors;T-Cell Depletion;T-Lymphocyte;Testing;Toxic effect;Transplantation;Veterans;Whole-Body Irradiation;abstracting;cytokine;design;dimer;experience;interleukin-23;leukemia;mutant;novel strategies

public_health_relevance:
Relevance to VA Mission: Every day large numbers of unfortunate veterans present with new cancers. In addition to the leukemias myelodysplasia and multiple myeloma other cancers could be amenable to stem cell transplantation. Admittedly there are some cancers that probably would not benefit from this therapy but the ability to achieve a revitalized immune system could be an important addition to their care. However as stated above the current risk of developing transplant related toxicities GVHD and overwhelming infection are too great to offer stem cell transplantation to patents with a wider variety of cancers. The proposed studies are to investigate a new approach to minimize GVHD


application_id: 7797702
project_number: 1I01BX000622-01
project_title: Mechanisms of Renal Lipid Accumulation in Diabetic Nephropathy
administering_ic: VA
award_notice_date: 2009-07-31
opportunity_number: BX-09-001
application_type: 1
activity_code: I01
ic: BX
serial_number: 000622
project_start_date: 2009-10-01
project_end_date: 2013-09-30
study_section: Nephrology[NEPH]
contact_pi_person_id: 3123430
contact_pi: TANNOCK, LISA R
other_pis: Not Applicable
congressional_district: 06
department: Unavailable
primary_duns: 018766373
primary_uei: QGWRFBMD3XT5
duns_number: 018766373
uei: QGWRFBMD3XT5
fips: US
organization_id: 481036
organization_name: VA MEDICAL CENTER - LEXINGTON, KY
organization_city: LEXINGTON
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 405022235
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2009-10-01
project_abstract:
DESCRIPTION (provided by applicant):   
  
Nephropathy is a major complication of both type 1 and type 2 diabetes which causes considerable morbidity and mortality. Despite best current treatments significant numbers of individuals with albuminuria progress to end stage renal disease. Although the mechanisms underlying this progression are not fully understood renal lipid and lipoprotein accumulation has been shown to accelerate the development of nephropathy. Renal lipid accumulation triggers an influx of inflammatory cells with subsequent development of glomerulosclerosis the characteristic lesion of diabetic nephropathy. Glomerulosclerosis is characterized by increased deposition of mesangial matrix including proteoglycans. Of particular interest the renal content of the small leucine-rich proteoglycan (SLRP) biglycan is increased in diabetes. TGF-2 which is elevated in diabetes and is known to be a key mediator of diabetic nephropathy development and progression also increases mesangial matrix deposition including increased expression of biglycan. Furthermore we have shown that TGF-2 increases the size and LDL binding affinity of renal proteoglycans. Thus increased renal proteoglycan (biglycan) synthesis induced by elevated TGF-2 in diabetes may be responsible for mediating the renal accumulation of lipoproteins. The overall goal of this grant is to test the hypothesis that renal lipid accumulation is mediated through interactions of lipoproteins with renal proteoglycans especially biglycan. This will be tested by comparing diabetic nephropathy in mice expressing proteoglycan-binding defective LDL with littermates expressing wildtype LDL. To determine if biglycan is the key proteoglycan responsible diabetic nephropathy will be compared between biglycan deficient and wildtype mice. The experiments outlined in this grant will provide direct in vivo experimental data identifying if proteoglycan mediated renal lipid accumulation contributes significantly to the development and progression of diabetic nephropathy and if the key proteoglycan is biglycan. This grant will also identify if biglycan serves as a natural inhibitor of TGF-2 in vivo. Thus the significance of this proposal is that it not only identifies a mechanism linking hyperlipidemia and diabetic nephropathy but also will identify novel targets to prevent or intervene in the development of diabetic nephropathy.  
  
PUBLIC HEALTH RELEVANCE:   
  
Potential Impact on Veterans Health Care: Veterans have very high prevalence of diabetes hyperlipidemia and diabetes complications including nephropathy. This research will either identify or exclude proteoglycans as a cause of renal lipid accumulation contributing to diabetic nephropathy. The grant will prove the concept in an animal model system and the results can be easily translated into clinical research. The ultimate goal is to identify novel targets for therapeutic intervention to decrease the morbidity and mortality of diabetic nephropathy.

public_health_relevance:
Potential Impact on Veterans Health Care: Veterans have very high prevalence of diabetes hyperlipidemia and diabetes complications including nephropathy. This research will either identify or exclude proteoglycans as a cause of renal lipid accumulation contributing to diabetic nephropathy. The grant will prove the concept in an animal model system and the results can be easily translated into clinical research. The ultimate goal is to identify novel targets for therapeutic intervention to decrease the morbidity and mortality of diabetic nephropathy.


application_id: 7797266
project_number: 1I01BX000610-01
project_title: Analysis of PTH and Dopamine Receptor Signaling in Proximal Tubules
administering_ic: VA
award_notice_date: 2009-07-28
opportunity_number: BX-09-001
application_type: 1
activity_code: I01
ic: BX
serial_number: 000610
project_start_date: 2009-10-01
project_end_date: 2013-09-30
study_section: Nephrology[NEPH]
contact_pi_person_id: 2262379
contact_pi: LEDERER, ELEANOR D
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2009-10-01
project_abstract:
DESCRIPTION (provided by applicant):   
  
Project Summary/Abstract  The goal of this project is to determine how the structural protein NHERF-1 sodium-hydrogen exchanger regulatory factor isoform 1 regulates the trafficking of Npt2a the type IIa sodium phosphate cotransporter to the apical membrane of the renal proximal tubule. The expression of Npt2a at the apical membrane is a critical regulatory step because the level of expression and function of Npt2a is the primary regulator of total body phosphate homeostasis. VA-funded research from this laboratory had revealed that the absence of NHERF-1 in a model of proximal renal tubule OK (opossum kidney) cells resulted in a marked decrease in Npt2a expression. The decrease in expression was due to two factors 1) decreased transcription of Npt2a and 2) decreased trafficking of Npt2a to the apical membrane. The mechanisms for the faulty trafficking of Npt2a have not been determined and are the subject of this proposal. Npt2a translated in the NHERF-deficient cells (OK-H) lacked a critical post-translational modification glycosylation. These proteins instead of trafficking to apical membrane accumulated in a perinuclear location. Inefficient apical membrane localization had previously been described in a mouse lacking expression of NHERF-1. Expression of a NHERF-1 construct that lacked a normal PDZ-2 domain did not allow trafficking of Npt2a to the apical membrane. NHERF-1 is a multiple PDZ domain protein which interacts with multiple signaling molecules and transporter proteins. A role for the PDZ-2 domain in Npt2a regulation had not been observed previously. Finally inhibition of SNARE (SNAP Receptor) protein interaction using a competing peptide introduced into OK cells blocked insertion of Npt2a into the apical membrane demonstrating that Npt2a is transported to the apical membrane via a vesicular transport mechanism.  The preliminary data suggested the hypothesis that NHERF-1 is essential for the forward trafficking of Npt2a from site of synthesis to the apical membrane. First the role of the post-translational modifications glycosylation and phosphorylation on Npt2a apical membrane trafficking will be examined by determining the cellular localization of transfected Npt2a constructs lacking the motifs required for these post translational modifications in a cell culture model. The glycosylation phosphorylation and ubiquitination states of Npt2a will be compared in NHERF-replete and NHERF-deficient OK cells and in wild type and NHERF-1 knock out mouse proximal tubule cells. The effect of inhibitors of glycosylation on the intracellular localization of Npt2a will be determined. Second the intracellular site or sites of interaction between NHERF-1 and Npt2a will be analyzed. Forward trafficking of Npt2a labeled by a GFP tag and by S35 methionine in OK cells and OK-H cells will be slowed by culture at 20 C. Analysis of Npt2a localization by confocal imaging density centrifugation and immunoelectronmicroscopy will be performed at sequential time points until the proteins are detected as biotinylated forms indicating appropriate insertion into the apical membrane. Trafficking of Npt2a in the two cell culture models will be compared. Proteins associated with Npt2a at each time point will be determined by immunoprecipitation and proteomic analysis. To define which steps in the forward trafficking are NHERF- dependent Npt2a trafficking will be compared in NHERF-replete and NHERF-deficient cells under conditions of traffic arrest: ezrin deficiency inhibition of SNARE interaction and CLC-5 (intracellular chloride channel CLC family isoform 5) deficiency. Third the sites on Npt2a and NHERF-1 critical for the NHERF-1 effect on Npt2a trafficking will be determined by mutational analysis of both proteins followed by both in vitro analysis of protein interaction and assessment of intracellular interaction by FRET methodology.  These studies will define where NHERF-1 acts in Npt2a forward trafficking define the sites on both proteins responsible for their interaction and yield mechanistic insights for this unique functional process.  
  
PUBLIC HEALTH RELEVANCE:   
  
Project Narrative  The goal of this project is to determine how the protein NHERF-1 sodium-hydrogen exchanger regulatory factor isoform 1 regulates the trafficking of Npt2a the type IIa sodium phosphate cotransporter to the apical membrane of the renal proximal tubule. The kidney regulates the total body content of phosphate through altering the expression of Npt2a at the apical membrane. Phosphate wasting can cause kidney stones and metabolic bone disease while phosphate retention is a risk factor for cardiovascular events common diseases in Veterans of all ages. Cardiovascular disease is the major cause of morbidity and mortality in the Veteran population. Kidney stones tend to affect individuals during their most productive years resulting in over $2 billion dollars of medical expenses in this country. Metabolic bone disease contributes substantially to the occurrence of hip fracture a disabling event with up to a 50% one year mortality. Thus understanding how the kidney regulates phosphate homeostasis has significant relevance to the health of Veterans of all ages.

project_terms:
Address;Affect;Age;Apical;CLC Gene;Cardiovascular Diseases;Carrier Proteins;Cell Culture Techniques;Cell Line;Cells;Centrifugation;Chloride Channels;Country;Data;Didelphidae;Disease;Dopamine Receptor;Event;Exhibits;Family;Fluorescence Resonance Energy Transfer;Funding;Genetic Transcription;Goals;Health;Hip Fractures;Homeostasis;Image;Immunoprecipitation;In Vitro;Individual;Kidney;Kidney Calculi;Knockout Mice;Label;Laboratories;Laboratory Research;Location;Medical;Membrane Microdomains;Membrane Protein Traffic;Membrane Transport Proteins;Metabolic Bone Diseases;Methionine;Methodology;Modeling;Molecular;Morbidity - disease rate;Mortality Vital Statistics;Mus;Mutation;Pathway interactions;Peptides;Phosphorylation;Physiologic pulse;Play;Population;Post-Translational Protein Processing;Process;Protein Analysis;Protein Isoforms;Proteins;Proteomics;Proximal Kidney Tubules;Receptor Signaling;Regulation;Renal function;Research;Research Personnel;Role;SNAP receptor;Signaling Molecule;Site;Staging;Structural Protein;Tertiary Protein Structure;Time;TimeLine;Transfection;Translating;Transport Vesicles;Tubular formation;Ubiquitination;Vesicle;Veterans;Work;abstracting;apical membrane;base;cardiovascular risk factor;caveolin 1;cold temperature;density;design;ezrin;glycosylation;in vitro Assay;inhibitor/antagonist;inorganic phosphate;insight;kidney cell;mutant;protein expression;receptor;research study;sodium-hydrogen exchanger regulatory factor;sodium-phosphate cotransporter proteins;solute;syntaxin binding protein 1;trafficking;wasting

public_health_relevance:
Project Narrative  The goal of this project is to determine how the protein NHERF-1 sodium-hydrogen exchanger regulatory factor isoform 1 regulates the trafficking of Npt2a the type IIa sodium phosphate cotransporter to the apical membrane of the renal proximal tubule. The kidney regulates the total body content of phosphate through altering the expression of Npt2a at the apical membrane. Phosphate wasting can cause kidney stones and metabolic bone disease while phosphate retention is a risk factor for cardiovascular events common diseases in Veterans of all ages. Cardiovascular disease is the major cause of morbidity and mortality in the Veteran population. Kidney stones tend to affect individuals during their most productive years resulting in over $2 billion dollars of medical expenses in this country. Metabolic bone disease contributes substantially to the occurrence of hip fracture a disabling event with up to a 50% one year mortality. Thus understanding how the kidney regulates phosphate homeostasis has significant relevance to the health of Veterans of all ages.


application_id: 7689007
project_number: 1I01BX000350-01
project_title: Alcohol Proteasome Dysfunction in Liver Injury
administering_ic: VA
award_notice_date: 2009-03-09
opportunity_number: BX-08-001
application_type: 1
activity_code: I01
ic: BX
serial_number: 000350
project_start_date: 2009-04-01
project_end_date: 2013-03-31
study_section: Gastroenterology[GAST]
contact_pi_person_id: 1902075
contact_pi: MCCLAIN, CRAIG J.
other_pis: Not Applicable
congressional_district: 03
department: Unavailable
primary_duns: 086765245
primary_uei: GXE7VBBR38K8
duns_number: 086765245
uei: GXE7VBBR38K8
fips: US
organization_id: 481037
organization_name: LOUISVILLE VA MEDICAL MEDICAL CENTER
organization_city: LOUISVILLE
organization_state: KY
organization_type: Independent Hospitals
organization_zip: 402061433
organization_country: UNITED STATES
arra_indicator: N
budget_start_date: 2009-04-01
project_abstract:
DESCRIPTION (provided by applicant):   
  
Studies from our VA-supported research and that of others demonstrate a critical role for the proinflammatory cytokine tumor necrosis factor (TNF) in the development and progression of alcoholic liver disease (ALD). Alcoholic liver disease is associated with increased oxidative stress and impaired hepatocyte 26S proteasome function. Our preliminary data demonstrate that inhibition of 26S proteasome function can sensitize hepatocytes to TNF cytotoxicity and higher doses of proteasome inhibitors directly induce hepatocyte death. Additionally this apoptotic death involves the induction of lysosomal membrane permeability (LMP) and is mediated by lysosomal proteases (cathepsins B and D). Based on the increasingly evident role of cathepsins as key downstream mediators of apoptosis and our preliminary results we postulate that chronic alcohol abuse with subsequent proteasome dysfunction leads to LMP and release of lysosomal cathepsins which induce hepatocyte cell death and further sensitize hepatocytes to TNF-mediated hepatotoxicity. Neutrophil infiltration is a hallmark of alcoholic hepatitis and infiltrating neutrophils are thought to play an etiologic role in the liver injury in ALD. We recently demonstrated that inhibition of proteasome function in hepatocytes induces and markedly enhances the TNF-inducible expression of the pro-inflammatory and neutrophil chemotactic cytokine Interleukin-8 (IL-8). It is our working hypothesis that chronic alcohol abuse mediated proteasome dysfunction is associated with increased hepatocyte IL-8 production which significantly contributes to neutrophil infiltration and hepatic injury in ALD. The overall objective of this proposal is to elucidate the cellular and molecular mechanism(s) involved in proteasome dysfunction-mediated hepatic injury associated with ALD. The specific objectives of this proposal are to: 1) Determine the effects of proteasome inhibition and ethanol on sensitization to TNF hepatotoxicity 2) Determine the role of lysosomal membrane permeability and cathepsins B and D in the proteasome dysfunction-mediated hepatocyte apoptosis and sensitization to TNF hepatotoxicity and 3) Determine the effects of proteasome inhibition and ethanol on the hepatocyte expression of the critical neutrophil chemotactic CXC cytokine IL-8. Potential Impact on Veterans Health: Alcoholic liver disease is a leading cause of death from liver disease in the United States and in the Veterans Administration. In studies from the Veterans Administration patients with cirrhosis and superimposed alcoholic hepatitis had > 60% mortality over a 4 year period of time with most of those deaths occurring in the first month. Thus the prognosis for this disease is more ominous than for many common types of cancer such as breast prostate and colon. The longterm goals of this laboratory are to define mechanism(s) for liver cell death in ALD that will provide new insights into this process and provide new areas for therapeutic intervention> This is especially important in ALD where there is no FDA-approved therapy. The potential clinical impact of this work is high.  
  
PUBLIC HEALTH RELEVANCE:   
  
 The overall objective of this proposal is to elucidate the cellular and molecular mechanism(s) involved in alcohol and proteasome dysfunction-mediated hepatic injury associated with alcoholic liver disease (ALD). We expect that the results of our study will provide critical molecular insights into the pathogenesis of ALD and provide a basis for clinically relevant paradigms for future development of potential therapeutic intervention(s) in patients with ALD.

project_terms:
26S proteasome;Affect;Alcohol abuse;Alcoholic Hepatitis;Alcoholic Liver Cirrhosis;Alcoholic Liver Diseases;Alcoholic beverage heavy drinker;Alcohols;Animals;Antioxidants;Apoptosis;Apoptotic;Area;Breast;Calpain;Cathepsins;Cathepsins B;Cause of Death;Cell Death;Cell Membrane Permeability;Cells;Cessation of life;Chronic;Cirrhosis;Clinical;Colon;Data;Development;Disease;Dose;Ethanol;Ethanol Metabolism;FDA approved;Functional disorder;Future;Genetic;Goals;Grant;Health;Hepatic;Hepatocyte;Hepatotoxicity;In Vitro;Inflammatory;Injury;Interleukin-8;Label;Laboratories;Liver;Liver diseases;Lysosomes;Mediating;Mediator of activation protein;Mitochondria;Modeling;Molecular;Morbidity - disease rate;Mortality Vital Statistics;Mus;Neutrophil Infiltration;Oxidative Stress;Pathogenesis;Pathway interactions;Patients;Peptide Hydrolases;Permeability;Plasma;Play;Polyubiquitin;Process;Production;Prostate;Proteasome Inhibition;Proteasome Inhibitor;Proteins;Proteolysis;Reactive Oxygen Species;Reduced Glutathione;Research;Research Support;Resistance;Role;Route;Signal Transduction;Stimulus;Therapeutic Intervention;Time;Tumor Necrosis Factor-alpha;Ubiquitin;United States;United States Department of Veterans Affairs;Veterans;Vitamin E;Work;alcohol effect;base;cancer type;chemokine;chronic alcohol ingestion;clinically relevant;cytokine;cytotoxicity;insight;monocyte;multicatalytic endopeptidase complex;neutrophil;outcome forecast;protein degradation;public health relevance;response

public_health_relevance:
The overall objective of this proposal is to elucidate the cellular and molecular mechanism(s) involved in alcohol and proteasome dysfunction-mediated hepatic injury associated with alcoholic liver disease (ALD). We expect that the results of our study will provide critical molecular insights into the pathogenesis of ALD and provide a basis for clinically relevant paradigms for future development of potential therapeutic intervention(s) in patients with ALD.


