INDUCIBLE MACROPHAGE SPECIFIC GENE EXPRESSION IN DISEASES
Jeanine M D´armiento
Columbia University Health Sciences, Columbia University Medical Center, New York, Ny 10032-3702
Grant 5RC1HL100384-02 from National Heart, Lung, And Blood Institute
Abstract: Develop transgenic animal models that are informative for understanding chronic inflammation in humans. Macrophages play a key role in the pathogenesis of multiple chronic inflammatory diseases. Studies using knockout mice have allowed investigators to examine the important role of macrophage-specific genes. However, these studies are limited to genes expressed in multiple tissues, as macrophage-specific knockout mice are not available. Since the loss of a gene may prevent the differentiation or migration of the monocyte prior to its actual arrival at the site of injury, the generation of an inducible, macrophage-specific knockout is preferable to study chronic inflammatory diseases. Our laboratory has developed a new transgenic mouse model targeting Cre recombinase expression in tissue macrophage, using the scavenger receptor A promoter (SRA-Cre). After its induction with tamoxifen, Cre will cleave any "floxed" gene in macrophages. Our model will also allow to knockout genes at any stage of the disease. The main goal of our proposal is to characterize this new conditional mouse model, using three aims. In the first aim, SRA-Cre mice will be crossed into a reporter line, and specificity and inducibility of Cre expression will be assessed in three models of lung injury (cigarette smoke exposure, asthma, and acute lung injury). In the second aim, SRA-Cre mice will be crossed into the apolipoprotein-E (Apoe) knockout background. Atherosclerotic plaques will be analyzed for expression of Cre in lesion macrophages and foam cells. This macrophage-specific Cre- expressing Apoe knockout model will be made available to laboratories studying the function of macrophages in atherosclerosis. In a third aim, we will generate of a conditional, macrophage- specific Tissue Inhibitor of Metalloproteinase-3 (Timp-3) knockout model. TIMP-3 is expressed in macrophage and plays an important role in inflammatory diseases. However, global Timp-3 knockout mice exhibited developmental abnormalities in the lung and the heart. We have generated a floxed Timp-3 model, which will be crossed into the SRA-Cre background. Utilizing this model, we will determine how the absence of TIMP-3 in macrophages affects lung injury due to cigarette smoke. Our new conditional, macrophage-specific targeting model will be a highly valuable tool, allowing researchers to precisely assess the roles and functions of genes expressed by macrophages during chronic inflammatory diseases. Macrophages play a key role in the pathogenesis of multiple chronic inflammatory diseases. In this proposal, we will characterize a newly developed conditional, macrophage-specific mouse model, where Cre recombinase is regulated by the scavenger receptor A promoter. Our model will allow to knockout genes of interest only in macrophages and at any stage of the disease. The major goal of our proposal is to generate a mouse model that will allow researchers to specifically and conditionally knockout genes of interest in macrophages during chronic inflammatory diseases. This model will be a highly valuable model, allowing researchers to precisely assess the roles and functions of genes expressed by macrophages during chronic inflammatory diseases. In addition, we propose to cross this model into the atherosclerosis-prone Apoe knockout model, to allow examination of macrophage genes in this important vascular disease. We will also examine the consequences of the absence of macrophage tissue inhibitor of metalloproteinases-3 in lung injury
Keywords: (Z)-2-[4(1, 2-diphenyl-1-butenyl)-phenoxyl]-N, N-dimethylethanamine; 1-p-beta-dimethylamino-ethoxyphenyl-trans-1, 2-diphenylbut-1-ene; APOE [{C0003595}]; Acute Pulmonary Injury; Affect; Animal Model; Animal Models and Related Studies; Apo-E; ApoE; ApoE knockout mouse; Apolipoprotein E; Apoptosis; Apoptosis Pathway; Area; Arterial Fatty Streak; Arthritis; Asthma; Atheroma; Atheromatous; Atheromatous degeneration; Atheromatous plaque; Atheroscleroses; Atherosclerosis; Atherosclerotic Cardiovascular Disease; Blood monocyte; Body Tissues; Bronchial Asthma; COAD; COPD; Cell Death, Programmed; Chronic; Chronic Obstructive Airway Disease; Chronic Obstructive Lung Disease; Cleaved cell; Development; Disease; Disorder; Ensure; Ethanamine, 2-(4-(1, 2-diphenyl-1-butenyl)phenoxy)-N, N-dimethyl-, (Z)-; Exhibits; Foam Cells; Gene Expression; Generations; Genes; Goals; Heart; Human; Human TIMP-3; Human Tissue Inhibitor of Metalloproteinase-3; Human, General; INFLM; Inflammation; Inflammatory; Injury; Investigation; Investigators; Knock-out; Knockout; Knockout Mice; Laboratories; Laboratory Study; Lesion; Lung; Lung Injury, Acute; Mammals, Mice; Man (Taxonomy); Man, Modern; Marrow monocyte; Mice; Mice, Knock-out; Mice, Knockout; Modeling; Murine; Mus; Null Mouse; Pathogenesis; Play; Principal Investigator; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Pulmonary Disease, Chronic Obstructive; Reporter; Research Personnel; Researchers; Respiratory System, Lung; Role; Site; Specificity; Staging; Streaks, Arterial Fatty; System; System, LOINC Axis 4; TAM; TIMP-3; Tamoxifen; Technology; Tissue Inhibitor of Metalloproteinase-3; Tissues; Transgenes; Transgenic Animals; Transgenic Mice; Vascular Diseases; Vascular Disorder; acetylated LDL receptor; acute lung injury; angiogenesis; arthritic; atheromatosis; atherosclerosis plaque; atherosclerotic lesions; atherosclerotic plaque; atherosclerotic vascular disease; blood vessel disorder; cigarette smoke; cleaved; disease/disorder; gene function; hTIMP; interest; knockout gene; lung injury; macrophage; migration; model organism; monocyte; mouse model; prevent; preventing; pulmonary; receptor, acetyl-LDL; recombinase; scavenger receptor; smoke of cigarettes; social role; tool; vulnerable plaque
Relevance: Narrative Macrophages play a key role in the pathogenesis of multiple chronic inflammatory diseases. In this proposal, we will characterize a newly developed conditional, macrophage-specific mouse model, where Cre recombinase is regulated by the scavenger receptor A promoter. Our model will allow to knockout genes of interest only in macrophages and at any stage of the disease. The major goal of our proposal is to generate a mouse model that will allow researchers to specifically and conditionally knockout genes of interest in macrophages during chronic inflammatory diseases. This model will be a highly valuable model, allowing researchers to precisely assess the roles and functions of genes expressed by macrophages during chronic inflammatory diseases. In addition, we propose to cross this model into the atherosclerosis- prone Apoe knockout model, to allow examination of macrophage genes in this important vascular disease. We will also examine the consequences of the absence of macrophage tissue inhibitor of metalloproteinases-3 in lung injury
Project start date: 2009-09-30
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: RFA-OD-09-003
5RC1HL100384-02 (2010): $492404
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Jeanine M D´armiento
GENETIC MODELS AND PHENOTYPING CORE
Jeanine M D´armiento, Assistant Professor Of Medicine
Columbia University Health Sciences, Columbia University Medical Center, New York, Ny 10032-3702
Abstract: This core is central for all transgenic and knockout experiments in this program project. Services that will be provided for by the core will include the generation of transgenic, knock-in and knockout animals, genotyping and transporting animals to Dr. Lederer for Project 3. The core will also provide support for the individual projects in the initial characterization of mouse models. The characterization of each mouse model will include careful analysis of the expression pattern of the transgene, and pathological and physiological changes, which occur as a result of transgene expression. In the past 5 years of the grant period the core managed over 300 cages of mice per year, assisted in pathological analysis of all new transgenic and knockout mice and performed physiological measurements on the mice. The core will provide support for the PPG in the following areas A. The preparation and maintenance of genetic mouse models B. Characterization of transgenic, knock-in and knock-out mice including preparation and analysis of tissue specimens from animals C. Baseline physiological measurements including hemodynamics and echocardiography D. Physiological measurements on transgenic and knockout mice for drug testing. E. Non-invasive testing for VT in free ranging WT and transgenic mice F. Electrophysiological studies on the in vivo murine heart. G. Electrocardiogram and blood pressure recordings
Keywords: Animals; Area; Blood Pressure; Body Tissues; Death, Sudden, Cardiac; ECG; EKG; Echocardiogram; Echocardiography; Electrocardiogram; Electrocardiography; Generations; Genetic; Genetic Models; Genotype; Grant; Heart; Individual; Knock-in; Knock-in Mouse; Knock-out; Knockout; Knockout Mice; Maintenance; Maintenances; Mammals, Mice; Measurement; Mice; Mice, Knock-out; Mice, Knockout; Mice, Transgenic; Models, Genetic; Molecular; Murine; Mus; Null Mouse; Pattern; Phenotype; Physiologic; Physiological; Preparation; Programs (PT); Programs [Publication Type]; Research Specimen; Services; Specimen; Testing; Tissues; Transgenes; Transgenic Mice; Transgenic Organisms; Transthoracic Echocardiography; base; drug detection; drug testing; experiment; experimental research; experimental study; heart sonography; hemodynamics; in vivo; knockout animal; mouse model; programs; research study; sound measurement; transgene expression; transgenic
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
5P01HL067849-09_9001 (2010): $399400
5P01HL067849-08_9001 (2009): $402084
Core B-Genetic Models And Phenotyping Core
Jeanine M D´armiento, Associate Professor
Columbia University Health Sciences
columbia University Medical Center
new York, Ny 100323702
Grant 5P01HL067849-079001 from National Heart, Lung, And Blood Institute IRG: HLBP
Abstract: This core is central for all transgenic and knockout experiments in this program project. Services that will be provided for by the core will include the generation of transgenic, knock-in and knockout animals, genotyping and transporting animals to Dr. Lederer for Project 3. The core will also provide support for the individual projects in the initial characterization of mouse models. The characterization of each mouse model will include careful analysis of the expression pattern of the transgene, and pathological and physiological changes, which occur as a result of transgene expression. In the past 5 years of the grant period the core managed over 300 cages of mice per year, assisted in pathological analysis of all new transgenic and knockout mice and performed physiological measurements on the mice. The core will provide support for the PPG in the following areas A. The preparation and maintenance of genetic mouse models B. Characterization of transgenic, knock-in and knock-out mice including preparation and analysis of tissue specimens from animals C. Baseline physiological measurements including hemodynamics and echocardiography D. Physiological measurements on transgenic and knockout mice for drug testing. E. Non-invasive testing for VT in free ranging WT and transgenic mice F. Electrophysiological studies on the in vivo murine heart. G. Electrocardiogram and blood pressure recordings
Keywords: biomedical facility, genetic model, genotype, laboratory mouse beta adrenergic receptor, gene targeting, heart electrical activity, hemodynamics, tachycardia echocardiography, electrocardiography, genetically modified animal, tissue /cell preparation
Jeanine M D´armiento
Columbia University Health Sciences, Columbia University Medical Center, New York, Ny 10032-3702
Abstract: for the generation of transgenic and knockout animals but the core will also support the individual projects in the initial characterization studies on the transgenic and knockout models. The characterization of each mouse model will include careful analysis of the expression pattern of the transgene and pathological and physiological changes, which occur as a result of transgene expression. The PI, Dr. Marks, Marx and Kass have collaborated over the past few years on the generation and characterization of the MK24=MinK-KVLQTl transgenic mouse (1) and FKBP12.6 knockout mice (2, 3). Physiological characterization of transgenic mice requires the technical expertise that is present in the laboratory of the core (4-6). Over the past three years systems have been set up in the laboratory to allow us to perform such studies. ´ The core will provide support for the PPG in the following areas ¿ A. The preparation and maintenance of transgenic and knockout mice B. Characterization of transgenic and knock-out mice including -preparation and analysis of tissue specimens from animals C. Baseline physiological measurements including hemodynamics and echocardiography D. Physiological measurements on transgenic and knockout mice for drug testing. ¿ Myocardial infarction and aortic banding when necessary F. Blood Pressure Measurements G. Maintenance of Cell CultureLines A major goal of the Animal Models and Tissue Culture Core is to develop and test animal models to provide models in which to examine allosteric interactions in ion channels. This Core will fill an essential and exciting role of the Columbia University PPG by facilitating the use of genetic models in animals using state-of-the-art techniques to make physiological measurements (eg,.ECG recording, EPS, echocardiograms and hemodynamic measurements) in mice. Finally, we will maintain all cell lines and develop stable transfected cell lines for the various constructs to be investigated. A stable cell line transfected with a wild-type expressing BK.channel has already been developed by Dr. Marx. Table 1. Mouse models used by each project Project 1-Karlin Project 2-Marks Project 3-Kass Project 4-Marx **(Tissue Culture) **(Tissue Culture) **(Tissue Culture) ***GuineaPigs J32AR* X X RyR2-S2809A* X X MK24* X FKBP12.6KO* X KCNQ1-L2M LAD Ligation X Model* Future models RyR2-ARVD2 X RyR2-CPVT X BKa knockout X RyR2-S2809D* X 32AR knockout X PHS 398/2590 (Rev. 05/01) Page 273 Continuation Format Page´
Budget start date: 1-AUG-2010
Budget end date: 31-JUL-2011
5P01HL081172-04_9001 (2010): $129222
5P01HL081172-03_9001 (2009): $219688
INDUCIBLE MACROPHAGE SPECIFIC GENE EXPRESSION IN DISEASES
Jeanine M D´armiento, Assistant Professor Of Medicine
Columbia University Health Sciences, Columbia University Medical Center, New York, Ny 10032-3702
Grant 1RC1HL100384-01 from National Heart, Lung, And Blood Institute
Abstract: Develop transgenic animal models that are informative for understanding chronic inflammation in humans. Macrophages play a key role in the pathogenesis of multiple chronic inflammatory diseases. Studies using knockout mice have allowed investigators to examine the important role of macrophage-specific genes. However, these studies are limited to genes expressed in multiple tissues, as macrophage-specific knockout mice are not available. Since the loss of a gene may prevent the differentiation or migration of the monocyte prior to its actual arrival at the site of injury, the generation of an inducible, macrophage-specific knockout is preferable to study chronic inflammatory diseases. Our laboratory has developed a new transgenic mouse model targeting Cre recombinase expression in tissue macrophage, using the scavenger receptor A promoter (SRA-Cre). After its induction with tamoxifen, Cre will cleave any "floxed" gene in macrophages. Our model will also allow to knockout genes at any stage of the disease. The main goal of our proposal is to characterize this new conditional mouse model, using three aims. In the first aim, SRA-Cre mice will be crossed into a reporter line, and specificity and inducibility of Cre expression will be assessed in three models of lung injury (cigarette smoke exposure, asthma, and acute lung injury). In the second aim, SRA-Cre mice will be crossed into the apolipoprotein-E (Apoe) knockout background. Atherosclerotic plaques will be analyzed for expression of Cre in lesion macrophages and foam cells. This macrophage-specific Cre- expressing Apoe knockout model will be made available to laboratories studying the function of macrophages in atherosclerosis. In a third aim, we will generate of a conditional, macrophage- specific Tissue Inhibitor of Metalloproteinase-3 (Timp-3) knockout model. TIMP-3 is expressed in macrophage and plays an important role in inflammatory diseases. However, global Timp-3 knockout mice exhibited developmental abnormalities in the lung and the heart. We have generated a floxed Timp-3 model, which will be crossed into the SRA-Cre background. Utilizing this model, we will determine how the absence of TIMP-3 in macrophages affects lung injury due to cigarette smoke. Our new conditional, macrophage-specific targeting model will be a highly valuable tool, allowing researchers to precisely assess the roles and functions of genes expressed by macrophages during chronic inflammatory diseases. Macrophages play a key role in the pathogenesis of multiple chronic inflammatory diseases. In this proposal, we will characterize a newly developed conditional, macrophage-specific mouse model, where Cre recombinase is regulated by the scavenger receptor A promoter. Our model will allow to knockout genes of interest only in macrophages and at any stage of the disease. The major goal of our proposal is to generate a mouse model that will allow researchers to specifically and conditionally knockout genes of interest in macrophages during chronic inflammatory diseases. This model will be a highly valuable model, allowing researchers to precisely assess the roles and functions of genes expressed by macrophages during chronic inflammatory diseases. In addition, we propose to cross this model into the atherosclerosis-prone Apoe knockout model, to allow examination of macrophage genes in this important vascular disease. We will also examine the consequences of the absence of macrophage tissue inhibitor of metalloproteinases-3 in lung injury
Keywords: (Z)-2-[4(1, 2-diphenyl-1-butenyl)-phenoxyl]-N, N-dimethylethanamine; 1-p-beta-dimethylamino-ethoxyphenyl-trans-1, 2-diphenylbut-1-ene; APOE [{C0003595}]; Acute Pulmonary Injury; Affect; Animal Model; Animal Models and Related Studies; Apo-E; ApoE; ApoE knockout mouse; Apolipoprotein E; Apoptosis; Apoptosis Pathway; Arterial Fatty Streak; Arthritis; Asthma; Atheroma; Atheromatous; Atheromatous degeneration; Atheromatous plaque; Atheroscleroses; Atherosclerosis; Atherosclerotic Cardiovascular Disease; Blood monocyte; Body Tissues; Bronchial Asthma; COAD; COPD; Cell Death, Programmed; Chronic; Chronic Obstructive Airway Disease; Chronic Obstructive Lung Disease; Cleaved cell; Development; Disease; Disorder; Ensure; Ethanamine, 2-(4-(1, 2-diphenyl-1-butenyl)phenoxy)-N, N-dimethyl-, (Z)-; Exhibits; Foam Cells; Gene Expression; Generations; Genes; Goals; Heart; Human; Human TIMP-3; Human Tissue Inhibitor of Metalloproteinase-3; Human, General; INFLM; Inflammation; Inflammatory; Injury; Investigation; Investigators; Knock-out; Knockout; Knockout Mice; Laboratories; Laboratory Study; Lesion; Lung; Lung Injury, Acute; Mammals, Mice; Man (Taxonomy); Man, Modern; Marrow monocyte; Mice; Mice, Knock-out; Mice, Knockout; Mice, Transgenic; Modeling; Murine; Mus; Null Mouse; Pathogenesis; Play; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Pulmonary Disease, Chronic Obstructive; Reporter; Research Personnel; Researchers; Respiratory System, Lung; Role; Site; Specificity; Staging; Streaks, Arterial Fatty; System; System, LOINC Axis 4; TAM; TIMP-3; Tamoxifen; Tissue Inhibitor of Metalloproteinase-3; Tissues; Transgenes; Transgenic Animals; Transgenic Mice; Vascular Diseases; Vascular Disorder; acetylated LDL receptor; acute lung injury; angiogenesis; arthritic; atheromatosis; atherosclerosis plaque; atherosclerotic lesions; atherosclerotic plaque; atherosclerotic vascular disease; blood vessel disorder; cigarette smoke; cleaved; disease/disorder; gene function; hTIMP; interest; knockout gene; lung injury; macrophage; migration; model organism; monocyte; mouse model; prevent; preventing; public health relevance; pulmonary; receptor, acetyl-LDL; recombinase; scavenger receptor; smoke of cigarettes; social role; tool; vulnerable plaque
Relevance: Narrative Macrophages play a key role in the pathogenesis of multiple chronic inflammatory diseases. In this proposal, we will characterize a newly developed conditional, macrophage-specific mouse model, where Cre recombinase is regulated by the scavenger receptor A promoter. Our model will allow to knockout genes of interest only in macrophages and at any stage of the disease. The major goal of our proposal is to generate a mouse model that will allow researchers to specifically and conditionally knockout genes of interest in macrophages during chronic inflammatory diseases. This model will be a highly valuable model, allowing researchers to precisely assess the roles and functions of genes expressed by macrophages during chronic inflammatory diseases. In addition, we propose to cross this model into the atherosclerosis- prone Apoe knockout model, to allow examination of macrophage genes in this important vascular disease. We will also examine the consequences of the absence of macrophage tissue inhibitor of metalloproteinases-3 in lung injury
Project start date: 2009-09-30
Project end date: 2011-08-31
Budget start date: 30-SEP-2009
Budget end date: 31-AUG-2010
PFA/PA: RFA-OD-09-003
1RC1HL100384-01 (2009): $476359
ALTERED MATRIX METALLOPROTEINASES IN HEART FAILURE
Jeanine M D´armiento, Associate Professor
Medicinecolumbia University Health Sciences
columbia University Medical Center
new York, Ny 100323702
Grant 5R01AG016994-04 from National Institute On Aging IRG: ZHL1
Abstract: Collagens are the major extracellular matrix component of the heart. The ECM provides structural integrity and tensile strength to the myocardium critical to maintaining the alignment of myocytes and myofibrils, and the overall geometry of the left ventricle. Fibrillar collagen degradation and changes in structural protein content have been postulated to contribute to myocyte slippage, wall thinning, and ventricular remodeling in heart failure. Interstitial collagen is highly resistant to degradation by all proteinases other than collagenases. Using the alpha-myosin heavy chain promoter we have created a transgenic mouse which constitutively expresses interstitial collagenase in the heart. Histological and functional analysis in these animals with disrupted matrix balance reveals myocyte and myofibrillar hypertrophy, altered collagen content, and ventricular dysfunction. This research proposal will investigate the role of the ECM and the balance of the protease-antiprotease system in cardiac function and in the pathophysiology of ventricular remodeling and the development of heart failure. The cardiac-collagenase mice will be subjected to hemodynamic and mechanical conditions that may impact on the cardiac remodeling in these animals. Experiments will be performed to identify changes in intracellular signaling which occur in the heart of the cardiac-collagenase mice. Studies will also determine if MMP-1 inhibitors are capable of halting the progression of the cardiac dysfunction seen in these animals. In conjunction with the animals studies, this proposal will investigate whether changes in the extracellular matrix proteins occur during reverse remodeling seen in human LVAD samples. Finally, this proposal will investigate the role of interstitial collagenase (MMP-13) in cardiac function and development through the generation of an MMP-13 knockout mouse
Keywords: collagenase, enzyme activity, heart cell, heart failure, muscle cell cardiovascular function, extracellular matrix, tissue inhibitor of metalloproteinase animal genetic material tag, laboratory mouse, transgenic animal
Project start date: 1998-09-30
Project end date: 2003-08-31
5R01AG016994-04 (2001): $298375
5R01AG016994-03 (2000): $298375
5R01AG016994-02 (1999): $298375
1R01AG016994-01 (1998): $298375
Sponsored Links Excellgen http://Excellgen.com
Jeanine M D´armiento, Associate Professor
Columbia University Health Sciences
columbia University Medical Center
new York, Ny 100323702
Grant 1P01HL081172-01A19001 from National Heart, Lung, And Blood Institute IRG: HLBP
Keywords: animal colony, biomedical facility, biomedical resource, genetically modified animal, laboratory mouse, tissue /cell culture blood pressure, cell line, drug screening /evaluation, echocardiography, genetic model, hemodynamics, histology, model design /development, myocardial infarction, tissue /cell preparation
Project start date: 2007-09-01
Project end date: 2012-07-31