Sabine Ehrt
Weill Medical College Of Cornell Univ
Project start date: 2010-12-03
Project end date: 2015-11-30
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Sabine Ehrt
CORE--BSL3 WET LAB AND VIVARIUM FACILITY
Sabine Ehrt, Assistant Professor
Weill Medical College Of Cornell Univ 1300 York Avenue New York, Ny 10021
Grant 1P01AI056293-019001 from National Institute Of Allergy And Infectious Diseases IRG: ZRG1
Abstract: The dual wet-lab and vivarium BSL-3 facility located on the 7th floor of the A building of the Weill Cornell Medical College is available to researchers approved to conduct in vitro or in vivo (animal) research with pathogens classified as BSL-3. The facility provides 900 nsf in a suite fully equipped for laboratory work, mouse housing, aerosol infection and tissue handling under conditions safe for work with BSL-3 organisms and compliant with CDC regulations. The facility is designed and operated to minimize the potential for environmental contamination with and personnel exposure to pathogenic microorganisms. This facility is essential for Projects 2, 3 and 4 in the Program Project, specifically, for work on Yersinia pestis and Mycobacterium tuberculosis. The 3 Pl s of these projects (Drs. Crystal, Nathan and Quadri) will represent 50% of the Pl s using the BSL3, and their work on these projects is estimated to represent 39% of the work in the BSL3.
Keywords: Mycobacterium tuberculosis, Yersinia pestis, biohazard control, biomedical facility, bioterrorism /chemical warfare, veterinary science
Project start date: 2003-07-01
Project end date: 2008-06-30
Macrophage Gene Expression: Impact Of M. Tuberculosis
Sabine Ehrt, Assistant Professor
Weill Medical College Of Cornell Univ 1300 York Avenue New York, Ny 10021
Grant 5R01HL068525-05 from National Heart, Lung, And Blood Institute IRG: ZHL1
Abstract: One third of the world s population is currently infected with Mycobacterium tuberculosis (Mtb) and every second, another person is newly infected with Mtb around the world. Each year an estimated 8 million people develop clinical disease and 1.87 million people die of TB. The primary host cell of Mtb is the macrophage and the Mtb-macrophage interaction is critical to every phase of Mtb s infectious cycle. Gene expression analysis (GEA) is the most effective technology currently available to record a quantitative picture of a cell s functional state, and therefore to compare cell states and types. The critical need for such a comparison in TB research is heightened when one takes into account the heterogeneity of macrophages by organ, species of origin and donor history. The goals of this study are to extend our understanding of macrophage biology relevant to tuberculosis (TB), and in so doing, to define the extent to which mouse macrophages can serve as surrogates for human macrophages. The latter question is key if functional genomics is used as a tool in TB research to test the course of infection by wild type and genetically modified Mtb in wild type and genetically modified mice. Specifically, we will extend our current use of high-density oligonucleotide microarrays, quantitative PCR and in situ hybridization (ISH) to analyze and compare the gene expression profiles of three distinct populations of primary cells mouse bone marrow macrophages (BMM), mouse pulmonary alveolar macrophages (PAM), and human PAM. These relatively pure macrophage populations will be compared with the heterogeneous mixture of all cells in mouse lung. Both human and mouse cells will come both from normal donors and individuals with active TB. Each cell population will be studied without further treatment, after infection in vitro with virulent Mtb, and/or after exposure to interferon gamma (IFNgamma), a cytokine critical for control of mycobacterial infection in both humans and mice. These comparisons will allow us to validate or qualify the use of mouse PAM, mouse BMM and human PAM as model systems for studying macrophage-Mtb interactions. The identification of genes regulated by Mtb in macrophages will generate hypotheses with respect to the role of these genes in pathogenesis. We will test at least one such hypothesis, that secretory leukocyte protease inhibitor (SLPI) plays a role in the pathogenesis of TB. Finally, we will share our GEA with others via the internet so that as many of the resulting hypotheses as possible can be rapidly and independently explored.
Keywords: Mycobacterium tuberculosis, gene expression, host organism interaction, macrophage, tuberculosis, alveolar macrophage, biological model, bone marrow, cell population study, cellular immunity, information dissemination, interferon gamma, lung, model design /development, oligonucleotide, pathologic process, protease inhibitor, Internet, human subject, in situ hybridization, laboratory mouse, lung lavage, microarray technology, patient oriented research, tissue /cell culture
Project start date: 2001-09-10
Project end date: 2007-07-31
5R01HL068525-05 (2005): $452920
5R01HL068525-04 (2004): $452920
5R01HL068525-03 (2003): $452920
5R01HL068525-02 (2002): $452920
1R01HL068525-01 (2001): $450488
DRUG TARGETS IN MTB GLUCONEOGENESIS
Sabine Ehrt
Department/ Educational Institution Type:
Grant 1R01AI092573-01 from National Institute Of Allergy And Infectious Diseases
Abstract: Mycobacterium tuberculosis (Mtb) causes latent infections that affect a third of the world´s population and active tuberculosis (TB) kills two million people every year. Chemotherapy of TB requires long treatment regimens and is complicated by the emergence of multi-drug resistant and extensively drug resistant Mtb strains. New drugs that shorten TB chemotherapy and cure drug resistant TB are urgently needed and their development requires a better understanding of the mechanisms used by this pathogen to persist in the host and cause disease. Pathogens need to acquire carbon from the host to establish and maintain an infection. Metabolic pathways used by Mtb during infections are therefore important for pathogenesis and can guide the development of new chemotherapies. Mtb is highly adapted to nutritionally stringent niches in the host and mounting evidence suggests that Mtb preferentially utilizes fatty acids during infections. Our preliminary data indicate that gluconeogenesis is essential for Mtb to grow and persist in immune-competent and immune- compromised mice. We will apply transcriptomic, metabolomic and biochemical approaches to determine the molecular consequences of inhibiting gluconeogenesis in vitro and in vivo, to gain mechanistic insight into the death associated with loss of gluconeogenic enzymes in vitro and during mouse infections, to identify inhibitors of gluconeogenesis and to validate gluconeogenic enzymes as potential drug targets. The proposed work will extend the limited knowledge on Mtb´s metabolism during infection and validate novel targets for chemotherapy. Tuberculosis is one of the world´s most devastating diseases. It is responsible for more than two million deaths and eight million new cases annually. Work outlined in this proposal will investigate metabolic adaptations that allow Mycobacterium tuberculosis to grow and persist within its host and to cause disease. It will help validate novel drug targets that might facilitate the development of new drugs against tuberculosis
Keywords: Abscission; Affect; Assay; Attenuated; attenuation; Bacteria; Bioassay; Biochemical; Biologic Assays; Biological Assay; Butyrates; C element; Carbohydrates; Carbon; Cause of Death; Cells; Cessation of life; chemotherapy; Chronic Phase; Citric Acid Cycle; Complement; Complement Proteins; Complex; Crystallographies; Crystallography; Data; Death; Development; Disease; disease/disorder; Disorder; disseminated TB; disseminated tuberculosis; Diversity Library; Drug Delivery; Drug Delivery Systems; Drug Resistance, Multiple; Drug Resistant Tuberculosis; Drug Resistant, Multiple; Drug Targeting; Drug Targetings; drug/agent; Drugs; Enzymes; Excision; Extirpation; Extreme drug resistant tuberculosis; Extremely drug resistant tuberculosis; Fatty Acids; Gene Action Regulation; Gene Expression Profile; Gene Expression Regulation; gene expression signature; gene product; Gene Regulation; Gene Regulation Process; Generalized Growth; Gluconeogenesis; Gluconeogenesis Inhibition; glucose biosynthesis; glyoxylate; Glyoxylate Transacetylase; Glyoxylates; Goals; Growth; Immune; In Vitro; in vivo; Infection; inhibitor; inhibitor/antagonist; insight; Intermediary Metabolism; Killings; Knowledge; Krebs Cycle; L-Malate glyoxylate-lyase (CoA-acetylating); latent infection; M. tb; M. tuberculosis; M.tb; M.tuberculosis; Malate Condensing Enzyme; Malate Synthase; Malate Synthetase; Mammals, Mice; measurement of metabolism; Measures; Mediating; Medication; Metabolic; Metabolic Pathway; Metabolic Processes; Metabolism; metabolomics; METBL; Mice; Molecular; Multi-Drug Resistance; multi-drug resistant; Multidrug Resistance; multidrug resistant; Murine; Mus; mutant; Mycobacterium tuberculosis; nitrosative stress; novel; ontogeny; Orthophosphate[{..}]oxaloacetate carboxy-lyase (phosphorylating); overexpression; pathogen; Pathogenesis; PEPCK; Pharmaceutic Preparations; Pharmaceutical Preparations; phosphoenolpyruvate carboxykinase; Phosphoenolpyruvate Carboxylase; Population; Proteins; Protocols, Treatment; public health relevance; Recombinants; Regimen; Removal; resection; Resistance; Resistance to Multi-drug; Resistance to Multidrug; Resistance to Multiple Drug; resistant; Resistant to multi-drug; Resistant to multidrug; Resistant to Multiple Drug; Resolution; RGM; Source; Starvation; Stress; Structure; sugar; Surgical Removal; TCA cycle; Testing; Tissue Growth; transcriptome; transcriptomics; Treatment Protocols; Treatment Regimen; Treatment Schedule; Tricarboxylic Acid Cycle; Tuberculosis; Tuberculosis, Drug Resistance; Tuberculosis, Drug Resistant; tuberculous spondyloarthropathy; Work; XDR-Tuberculosis
Relevance: Tuberculosis is one of the world´s most devastating diseases. It is responsible for more than two million deaths and eight million new cases annually. Work outlined in this proposal will investigate metabolic adaptations that allow Mycobacterium tuberculosis to grow and persist within its host and to cause disease. It will help validate novel drug targets that might facilitate the development of new drugs against tuberculosis
Project start date: 2010-12-03
Project end date: 2015-11-30
Budget start date: 3-DEC-2010
Budget end date: 30-NOV-2011
PFA/PA: PA-10-067
1R01AI092573-01 (2011): $626426
PH Homeostasis In Mycobacterium Tuberculosis
Sabine Ehrt, Associate Professor
Microbiology And Immunologyweill Medical College Of Cornell Univ
Grant 1R01AI081725-01 from National Institute Of Allergy And Infectious Diseases IRG: HIBP
Abstract: M. tuberculosis (Mtb) causes latent infections that affect a third of the world´s population and active tuberculosis kills two million people every year. Chemotherapy of tuberculosis requires long treatment regimens and is complicated by the emergence of multi-drug resistant and extensively drug resistant Mtb strains. New drugs that shorten TB chemotherapy and cure drug resistant TB are urgently needed. Mtb encounters an acidic pH within phagosomes of interferon-3 activated macrophages and must prevent excessive entry of protons into its cytosol. One pathway important for pH homeostasis and virulence of Mtb depends on the membrane-associated serine hydrolase Rv3671c. The goals of this proposal are to determine the molecular mechanisms by which Rv3671c protects Mtb from intracellular acidification and contributes to virulence. Our specific aims are I. To determine why Rv3671c is required for resistant of Mtb against acid. We will investigate the mechanism by which this serine hydrolase controls pH and acid resistance. II. To determine why Rv3671c is required for persistence in activated macrophages and mice. We will investigate if Rv3671c affects phagosome maturation in IFN-3 activated macrophages. We will use genetically altered mice to determine if Rv3671c protects against host defense mechanisms, which act synergistically with phagosome acidification. III. To identify small molecules that interfere with pH homeostasis in Mtb. We will use a high- throughput screen to identify such inhibitors, which will help to investigate the biology of intrabacterial pH homeostasis in Mtb and facilitate the development of new drugs against tuberculosis. Tuberculosis (TB) is one of the world´s most devastating diseases. It is responsible for more than two million deaths and eight million new cases annually. Work outlined in this proposal will investigate virulence mechanisms that allow Mtb to persist within its host and cause disease. It will help identify novel drug targets that might facilitate the development of new drugs against tuberculosis
Project start date: 2009-03-01
Project end date: 2014-02-28
Conditional Expression Of Mycobacterial Genes
Sabine Ehrt, Associate Professor
Weill Medical College Of Cornell Univ 1300 York Avenue New York, Ny 10021
Grant 5R01AI063446-02 from National Institute Of Allergy And Infectious Diseases IRG: HIBP
Abstract: Among communicable diseases, tuberculosis is the second leading cause of death worldwide. New antimycobacterial drugs that allow a shortened but effective chemotherapy, that target replicating as well as non-replicating mycobacteria, and are active against multidrug-resistant Mycobacterium tuberculosis (Mtb) are urgently needed. Recent advances in functional genomics identified a number of essential genes that represent potential targets for the development of novel antibiotics. However, essential genes are difficult to study and it is unknown which of them are required during infection, in particular during the chronic phase of tuberculosis. To address this, we propose to develop mycobacterial tet-on and tet-off systems that allow to regulate gene expression of Mtb in vitro and in vivo. In Aim I we will optimize tet-on and develop tet-off systems for efficient gene induction and silencing. In Aim II we will use these systems to generate conditional knockout strains and analyze the role of glycolysis and gluconeogenesis during growth of Mtb with defined carbon sources. In Aim III we will use tet systems to regulate gene expression in Mtb during growth in mice and determine if glycolysis and gluconeogenesis are required for in vivo growth of Mtb. The experiments proposed here will (i) provide the TB-research community with a molecular tool that allows expression of mycobacterial genes at different time points during an infection (ii) provide a molecular tool for the validation of new drug targets; and (iii) provide insights into the metabolic adaptations required for Mtb to grow within its host. Many of the experiments focus on generally conserved genes and will, therefore, lead to insights that may be relevant to other pathogens and diseases. Lay Summary Tuberculosis (TB) is one of the world s most devastating diseases. It is responsible for more than two million deaths and eight million new cases annually. Work outlined in this proposal will develop molecular tools to aid the understanding of how Mycobacterium tuberculosis grows and persists within its host. It will help identify and validate novel drug targets that might lead to the development of antibiotics against replicating and non-replicating Mycobacterium tuberculosis and thus shorten anti-tuberculosis drug therapy.
Keywords: gene, gene expression, gluconeogenesis, glycolysis, tuberculosis, Mycobacterium, Mycobacterium tuberculosis, antibiotic, carbon, chemotherapy, communicable disease, community, death, fatty acid, functional /structural genomics, gene induction /repression, gene targeting, glucose, glycerol, infection, insight, lead, macrophage, mutant, performance, phenotype, reporter gene, role, tetracycline, therapy
Project start date: 2006-02-15
Project end date: 2011-01-31
5R01AI063446-02 (2007): $407820
1R01AI063446-01A2 (2006): $409800
Sponsored Links Excellgen http://Excellgen.com
Sabine Ehrt
Weill Medical College Of Cornell Univ
Project start date: 2009-03-01
Project end date: 2014-02-28
Conditional Expression Of Mycobacterial Genes
Sabine Ehrt, Associate Professor
Microbiology And Immunologyweill Medical College Of Cornell Univ
Grant 5R01AI063446-04 from National Institute Of Allergy And Infectious Diseases IRG: HIBP
Keywords: gene, gene expression, gluconeogenesis, glycolysis, tuberculosis Mycobacterium, Mycobacterium tuberculosis, antibiotic, carbon, chemotherapy, communicable disease, community, death, fatty acid, functional /structural genomics, gene induction /repression, gene targeting, glucose, glycerol, infection, insight, lead, macrophage, mutant, performance, phenotype, reporter gene, role, tetracycline, therapy
Project start date: 2006-02-15
Project end date: 2011-01-31