Richard M Ransohoff
Cleveland Clinic Lerner Col/med-cwru
Project start date: 1994-05-01
Project end date: 2014-01-31
Sponsored Links Excellgen http://Excellgen.com
CHEMOKINES IN CNS INFLAMMATION
Richard M Ransohoff, Staff
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Grant 5R01NS032151-15 from National Institute Of Neurological Disorders And Stroke
Abstract: The current application focuses on CXCR2, a chemokine receptor present both on leukocytes and parenchymal central nervous system (CNS) cells. Our preliminary findings and recent reports demonstrate that CXCR2 on circulating leukocytes is decisive with regard to the outcome of EAE or cuprizone-mediated demyelination. Our recent findings show further that CXCR2 on CNS cells also regulates myelin repair. CXCR2 is the first chemokine receptor which promotes myelin destruction and hinders myelin repair. Our studies will address the fundamental role of CXCR2 in demyelination and remyelination, by performance of the following Specific Aims Aim 1. We will define the pathways by CXCR2 expression by leukocytes contributes to cuprizone- mediated demyelination. Hypothesis CXCR2 promotes demyelination through recruitment of monocytes to cuprizone lesions. The vast majority of hematogenous inflammatory cells in cuprizone lesions are monocyte- derived macrophages, leading us to suggest that CXCR2 acts through macrophage recruitment. The results of our proposed experiments will define how myeloid-cell CXCR2 contributes to this process. Aim 2. We will use an in-vitro model of demyelination-remyelination to address how the presence of CXCR2 impairs remyelination. Preliminary results showed that antibody-mediated blockade or genetic absence of CXCR2 led to accelerated remyelination of brain slices in-vitro. Hypothesis The presence of CXCR2 on oligodendrocyte progenitor cells impairs their proliferation and differentiation in demyelinated lesions. This in-vitro model recapitulates remyelination in-vivo, particularly those aspects which take place within the demyelinated lesion. This convenient, manipulable model will be used to examine the cellular and molecular basis by which CXCR2 hinders remyelination. Aim 3. We will define CNS cells whose expression of CXCR2 leads to inefficient remyelination. To address CXCR2 function in the CNS, we generated a conditional CXCR2 allele. Hypothesis CXCR2 impairs remyelination by slowing accumulation of oligodendrocyte progenitors in lesions and hindering their proliferation and differentiation. Mice harboring a conditional CXCR2 allele will be crossed with available mice expressing cre-ER(T2) in individual cell types, allowing efficient lineage-specific, inducible deletion. We will analyze cuprizone-mediated demyelination and remyelination to probe how CXCR2 expression in CNS cells regulates this process. These studies will provide essential data for designing clinical trials of CXCR2 blockade in MS. This grant focuses on CXCR2, a cellular receptor present both on inflammatory white blood cells and brain cells. CXCR2 both promotes myelin destruction and hinders myelin repair. Our studies will address the fundamental role of CXCR2 in demyelination and remyelination, by using novel models including in-vitro brain slices and genetically modified mice, allowing the receptor to be removed either from inflammatory white blood cells or from CNS cells. Ongoing clinical trials for pulmonary disease use drugs that block CXCR2 and our studies will provide essential data for designing clinical trials of CXCR2 blockade in MS
Keywords: Address; Affect; Alleles; Allelomorphs; Antibodies; Biscyclohexa Oxaldihydrazone; Blood Neutrophil; Blood Polymorphonuclear Neutrophil; Blood Segmented Neutrophil; Blood leukocyte; Blood monocyte; Body Tissues; Bone Marrow; Brain; CDw128b; CDw128b Antigens; CMKAR2; CNS Diseases; CNS disorder; CXC Chemokine Receptor 2; CXCR2; CXCR2 Protein; Cell Communication and Signaling; Cell Signaling; Cells; Central Nervous System; Central Nervous System Diseases; Central Nervous System Disorders; Chemokine (C-X-C) Receptor 2; Chimera; Chimera organism; Clinical Trials; Clinical Trials Design; Clinical Trials, Unspecified; Complement; Complement Proteins; Complex; Cuprizone; Cytokines, Chemotactic; Data; Demyelinations; Drug usage; EAE; Encephalomyelitis; Encephalomyelitis, Allergic; Encephalon; Encephalons; Ethanedioic acid, bis(cyclohexylidenehydrazide); Experimental Allergic Encephalitis; Experimental Allergic Encephalomyelitis; Experimental Autoimmune Encephalitis; Experimental Autoimmune Encephalomyelitis; GRO/MGSA Receptor; Genetic; Glia; Glial Cells; Grant; Hematogenous; Heterophil Granulocyte; High Affinity Interleukin 8 Receptor Type B; Homologous Chemotactic Cytokines; IL-8RB; IL-8Rbeta; IL8 Receptor Type 2; IL8R2; IL8RB; IL8RB gene; INFLM; In Vitro; Individual; Inflammation; Inflammatory; Intercrines; Interleukin 8 Receptor Beta; Interleukin 8 Receptor Type 2; Interleukin-8 Receptor Type B; Interleukin-8 Receptors B; Interleukin-8B Receptor; Intracellular Communication and Signaling; Kolliker`s reticulum; Lesion; Leukocytes; Ligands; Liposomal; Liposomes; Lung diseases; Lysolecithins; Lysophosphatidylcholines; MS (Multiple Sclerosis); Mammals, Mice; Marrow Neutrophil; Marrow leukocyte; Marrow monocyte; Mediating; Mice; Modeling; Molecular; Mother Cells; Multiple Sclerosis; Murine; Mus; Myelin; Myeloencephalitis; Myelogenous; Myeloid; Myeloid Cells; Nerve Cells; Nerve Unit; Nervous System, Brain; Nervous System, CNS; Neural Cell; Neuraxis; Neurocyte; Neuroglia; Neuroglial Cells; Neurons; Neutrophilic Granulocyte; Neutrophilic Leukocyte; Non-neuronal cell; Oligodendrocytes; Oligodendrocytus; Oligodendroglia; Oligodendroglia Cell; Outcome; Pathway interactions; Performance; Peripheral; Phenotype; Polymorph; Polymorphonuclear Cell; Polymorphonuclear Leukocytes; Polymorphonuclear Neutrophils; Process; Progenitor Cells; Protocol; Protocols documentation; Pulmonary Diseases; Pulmonary Disorder; Receptor Protein; Receptors, CXCR2; Relative; Relative (related person); Reporting; Research; Resistance; Respiratory Disease; Respiratory Disorder; Respiratory System Disease; Respiratory System Disorder; Reticuloendothelial System, Bone Marrow; Reticuloendothelial System, Leukocytes; Role; SIS cytokines; Sclerosis, Disseminated; Signal Transduction; Signal Transduction Systems; Signaling; Slice; Stem cells; Tissues; White Blood Cells; White Cell; autoimmune encephalomyelitis; base; biological signal transduction; brain cell; cell type; chemoattractant cytokine; chemokine; chemokine receptor; clinical investigation; drug use; experiment; experimental research; experimental study; in vitro Model; in vivo; insight; insular sclerosis; lung disorder; macrophage; monocyte; nerve cement; neuronal; neutrophil; novel; pathway; progenitor; public health relevance; receptor; repair; repaired; research study; resistant; response; social role; white blood cell; white blood corpuscle
Relevance: This grant focuses on CXCR2, a cellular receptor present both on inflammatory white blood cells and brain cells. CXCR2 both promotes myelin destruction and hinders myelin repair. Our studies will address the fundamental role of CXCR2 in demyelination and remyelination, by using novel models including in-vitro brain slices and genetically modified mice, allowing the receptor to be removed either from inflammatory white blood cells or from CNS cells. Ongoing clinical trials for pulmonary disease use drugs that block CXCR2 and our studies will provide essential data for designing clinical trials of CXCR2 blockade in MS
Project start date: 1994-05-01
Project end date: 2014-01-31
Budget start date: 1-FEB-2010
Budget end date: 31-JAN-2011
PFA/PA: PA-07-070
5R01NS032151-15 (2010): $306002
ASTROCYTE SYNTHESIS OF CHEMOTACTIC CYTOKINES IN EAE
Richard M Ransohoff, Director
Cleveland Clinic Foundation 9500 Euclid Ave. Cleveland, Oh 44195
Grant 5R01NS032151-04 from National Institute Of Neurological Disorders And Stroke IRG: NEUC
Abstract: Inflammatory cell recruitment to the central nervous system (CNS) is a cardinal feature of numerous pathological processes, including multiple sclerosis (MS). Despite recent progress, the soluble signals that attract inflammatory cells from perivascular accumulations into the CNS parenchyma remain obscure. The present proposal derives from experience in our laboratories and is based on the hypothesis that a specific class of chemoattractant cytokines termed chemokines is synthesized by astrocytes in response to inflammatory stimuli; we propose that astrocyte-derived chemokines in turn function to recruit inflammatory cells into the CNS parenchyma. This hypothesis will be addressed by performance of two Specific Aims Al. We will establish the relationship between chemokine gene expression and CNS inflammation in informative model systems. Three models will be studied, each with specific advantages 1) EAE of SJL/J mice actively immunized with proteolipid protein (PLP) encephalitogens; 2) EAE of Lewis rats, after adoptive transfer of myelin basic protein (MBP)-specific or myelin oligodendroglial glycoprotein (MOG)-specific T-cell lines; 3) Traumatic brain injury in neonatal and adult mice. We will assess chemokine transcript levels by a sensitive, semi-quantitative reverse transcription/polymerase chain reaction (RT/PCR) assay. Subsequently, we will establish the relationship between specific histopathologic alterations and localized chemokine production in CNS tissues, assayed by in situ hybridization (ISH). A2. We will evaluate regulation of chemokine gene expression in astrocytic cells in vitro and murine CNS in vivo. Our preliminary work showed that CNS expression of chemokine mRNAs was limited to astrocytes in mice with EAE. To determine the mechanism of this chemokine gene expression, we will address the cytokine regulators of IP-10 and JE/MCP-l transcription in human astrocytic and monocytic cells in vitro and in murine CNS cells in vivo 1) Regulation of chemokine gene expression in human astrocytic and monocytic cells will be compared. 2) Chemokine gene expression in the murine CNS will be determined by RT/PCR and ISH in vivo after intravenous injection of pro-inflammatory cytokines IFNgamma and TNFalpha. The production of chemoattractant cytokines by astrocytes provides insight into the role of endogenous neural cells in amplifying inflammatory processes. Research proposed here will enhance our understanding of varied pathologic processes and suggest potential therapeutic strategies for disorders limited to the CNS.
Keywords: astrocyte, chemokine, experimental allergic encephalomyelitis, interferon gamma, neuroimmunomodulation, tumor necrosis factor alpha, brain injury, gene deletion mutation, gene induction /repression, genetic promoter element, inflammation, monocyte, myelin basic protein, myelin glycoprotein, myelin proteolipid, protein biosynthesis, transcription factor, trauma, human tissue, in situ hybridization, laboratory mouse, laboratory rat, newborn animal, polymerase chain reaction, tissue /cell culture
Project start date: 1994-05-01
Project end date: 1998-04-30
5R01NS032151-04 (1997): $162921
5R01NS032151-02 (1995): $150630
Chemokines In CNS Inflammation
Richard M Ransohoff, Director
Cleveland Clinic Lerner Col/med-cwru P84 Cleveland, Oh 44195
Grant 5R01NS032151-13 from National Institute Of Neurological Disorders And Stroke IRG: ZRG1
Abstract: Chemokines and their G-protein coupled receptors constitute a unique ligand-receptor system that governs leukocyte migration and effector function. Chemokines are present in most multicellular organisms, to mediate organ patterning during development. In mammals, chemokines also regulate the function of a sophisticated, flexible host-defense system. The central nervous system (CNS) lesions of multiple sclerosis (MS), an inflammatory demyelinating disease, contain specific populations of activated leukocytes, including monocytes, macrophages, microglia and lymphocytes. Understanding the molecular determinants of leukocyte trafficking to the CNS will promote progress towards effective treatment of MS. Data obtained in the previous round of funding supported our core hypothesis that individual chemokine/receptor pairs are selectively involved in the recruitment and activation of specific leukocyte populations in the CNS. Using experimental autoimmune encephalomyelitis (EAE) as a model system, we propose to examine our hypothesis in detail. We have carefully selected genetically-modified mice that will allow us to examine chemokine signaling in the critical cell populations implicated in EAE and MS. Using these powerful reagents, we intend to clarify how chemokines and their receptors govern the CNS recruitment and activation of diverse cell populations that take part in EAE monocytes, lymphocytes, astrocytes, NK cells and microglia. The Specific Aims address the following questions 1. How do MCP- l/CCL2 and CCR2 govern cell recruitment and activation in the CNS? 2. How do fractalkine/CX3CL1 and CX3CR1 regulate the recruitment and activation of resident microglia and infiltrating cells in the CNS of mice with EAE? 3. How do CXCR3 and its ligands govern the accumulation and distribution of T-cells in the CNS of mice with EAE?
Keywords: astrocyte, cell migration, chemokine, cytokine receptor, experimental allergic encephalomyelitis, gene expression, lymphocyte, microglia, monocyte, natural killer cell, receptor expression, IP 10 protein, cell motility, cytotoxic T lymphocyte, helper T lymphocyte, macrophage inflammatory protein, monocyte chemoattractant protein 1, cell line, genetically modified animal, immunocytochemistry, laboratory mouse, polymerase chain reaction, tissue /cell culture
Project start date: 1994-05-01
Project end date: 2008-03-31
5R01NS032151-13 (2006): $354836
5R01NS032151-12 (2005): $363375
5R01NS032151-10 (2003): $351500
5R01NS032151-08 (2001): $206836
5R01NS032151-07 (2000): $200810
5R01NS032151-06 (1999): $194962
ASTROCYTE SYNTHESIS OF CHEMOTACTIC CYTOKINES IN EAE
Richard M Ransohoff, Director
Cleveland Clinic Foundation 9500 Euclid Ave. Cleveland, Oh 44195
Grant 5R01NS032151-03 from National Institute Of Neurological Disorders And Stroke IRG: NEUC
Project start date: 1994-05-01
Project end date: 1998-04-30
5R01NS032151-03 (1996): $156655
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Richard M Ransohoff
BETA R1 GENE: MODEL SYSTEM TO STUDY IFN BETA SIGNALING
Richard M Ransohoff, Director
Cleveland Clinic Foundation 9500 Euclid Ave. Cleveland, Oh 44195
Grant 5P01CA062220-100003 from National Cancer Institute
Abstract: Type I interferons include more than l0 subtypes of IFN-alpha, as well as IFN-beta, IFN-tau and IFN-omega. These related cytokines all signal through the IFN alpha/beta receptor, a complex composed of two transmembrane components (IFNAR-l/2) and two nonreceptor protein tyrosine kinases, TYK2 and JAK1. It has recently become clear that diverse signals can be generated by engagement of this receptor complex with its various ligands. Two distinctive outcomes of treatment with IFN-beta have been described inducible transcription of a gene termed beta-R1, and assembly of a stable complex of IFN-beta with the two receptor chains. We propose that the induction of beta-Rl requires both common and unique signaling through the IFNAR-l/2 complex. This hypothesis is based on our preliminary results, which demonstrate that beta-RI transcription is dependent on all components needed to form the transcription factor ISGF3, but is atypically sensitive to phosphoinositol-3 kinase (PI3K) inhibitors and exhibits an unusual requirement for catalytically active TYK2. This research proposal addresses the hypothesis that 13-RI transcription depends on conformation-dependent signaling through the IFNAR-1/2 complex in response to IFN-beta as compared with IFN-alpha. This signaling pathway acts through catalytic TYK2 and PI3K, as well as ISGF3. The specific aims are to l) characterize IFN-beta signaling to the beta3-Rl promoter; 2) generate cells selectively unresponsive to IFN-beta and 3) determine gene expression selectively regulated by IFN-beta. Performance of these aims will provide insight into IFN-beta-specific signaling and help clarify the functional roles of accessory components, such as PI3K, in type I IFN response pathway.
Keywords: biological signal transduction, cytokine receptor, gene expression, genetic regulation, genetic regulatory element, genetic transcription, interferon beta, protein tyrosine kinase, receptor binding, JAK kinase, gene induction /repression, genetic promoter element, interferon alpha, transcription factor, tissue /cell culture
Mentored Research: Chemokine Regulation On CNS Inflammation In MS
Richard M Ransohoff, Director
Cleveland Clinic Lerner Col/med-cwru P84 Cleveland, Oh 44195
Grant 5K24NS051400-02 from National Institute Of Neurological Disorders And Stroke IRG: NST
Abstract: This revised proposal gives equal attention to the priorities of mentoring and the research plan. The research plan is directed to understanding the anatomic and functional aspects of chemokine receptor expression in the lesions of multiple sclerosis (MS) a disease, which is the most prevalent primary neurological cause of disability in the United States. Our data show that chemokine receptors are selectively expressed by infiltrating cells (monocytes and T cells) in MS lesions, with different patterns, that vary according to lesion character. These findings prompted us to address the following questions First, how does chemokine receptor expression correlate with MRI-characteristics of MS lesions, as determined in a unique series of tissues subjected to MRI/pathological correlations? Second, is there heterogeneous expression of chemokines and their receptors, in different lesions of MS within a single brain? We will examine these questions by immunohistochemical studies of tissue sections from MS patients and appropriate controls. We will test hypotheses derived from these studies using functional analysis, using a novel in vitro model of the blood brain barrier (BBB). The plan entails new directions for the PI, who will become familiar with imaging research techniques. Further, in the effort to identify biomarkers that indicate which pathological patterns are represented by MRI lesion characteristics in vivo, will require familiarity with a diverse range of clinical research methods. Together, these new directions necessitate a period of intense research focus under the K24 Award. This program will also allow for expanded mentoring activities, for beginning clinician investigators. The mentoring component of this proposal includes progression from descriptive immunohistochemical analysis of MS tissue sections, through hypothesis generation based on the descriptive data, culminating in hypothesis testing, using the BBB model system. The mentorship will be enriched by participation of an advisory team including other clinical and basic researchers as well as biostatistical support. Formal classwork and informal guidance are integrated in the mentoring plan, which will result in development of clinician-scientists who are enabled to use cutting edge research techniques to elucidate human disease.
Keywords: chemokine, chemokine receptor, inflammation, antibody, attention, base, biomarker, blood, blood brain barrier, brain, cell, central nervous system, clinical trial, evolution, human, insight, leukocyte, microglia, model, monocyte, multiple sclerosis, receptor, receptor expression, sclerosis, sectioning, small molecule, tissue, clinical research
Project start date: 2006-02-08
Project end date: 2011-01-31
5K24NS051400-02 (2007): $169668
1K24NS051400-01A1 (2006): $169668
Chemokines In A Novel Murine Model Of DTH In The Brain
Richard M Ransohoff, Director
Cleveland Clinic Lerner Col/med-cwru P84 Cleveland, Oh 44195
Grant 7R03TW006012-03 from Fogarty International Center IRG: ICP
Abstract: This research will be done primarily in Poland as an extension of NIH grant # 1 R01 NS 32151 "Chemokines in CNS inflammation." Leukocyte infiltration into the central nervous system (CNS) is a key event in many neuroimmunologic diseases. The recruitment of lymphocytes and macrophages into the brain is likely the result of chemokine (chemoattractant cytokine) expression in the CNS. In the parent grant we analyze how specific pairs of chemokines and their receptors govern recruitment of inflammatory cells to the CNS during animal model of multiple sclerosis (MS) - experimental autoimmune encephalomyelitis (EAE). In this proposal, we would like to extend those studies and analyze the role of chemokines and chemokine receptors in a newly described focal model of MS-like CNS lesions induced by intracerebral injection of bacillus Calmette-Guerin (BCG). This model is an example of brain delayed type hypersensitivity (DTH) reaction and is characterized by the presence of a single mononuclear inflammatory focus in brain striatum. Unlike EAE brain, the DTH model is induced by non-CNS antigen (BCG) and allows observation of bystander damage to the brain triggered by inflammatory reaction. Similar types of CNS damage is postulated for MS pathogenesis. This research will concentrate on the following three Specific Aims 1) Development and characterization of the murine model of brain DTH reaction. Initially, murine model of brain DTH reaction will be described. Originally this model was described in rats a few years ago. Mouse DTH model will enable us to use several murine reagents not available for rats and use genetically modified animals in the future. 2) Analysis of chemokines and chemokine receptor expression in brain DTH model. Once the histopathology of a new model is described, we will address the potential role of chemokines in its development. Expression of chemokines and chemokine receptors in the brain during development of DTH lesion will be analyzed. This project will be concluded with Aim 3) Prevention and treatment of brain DTH model with chemokine inhibitors. Some chemokines and their receptors are most likely upregulated in the brain during BCG-induced model of neuroinflammation. This observation may lead to modulation of this pathology with anti-chemokine strategy. Upregulated chemokines will also be targeted in Aim 3. Research proposed here will complement ongoing studies by the parent grant without overlap. Results of those studies may suggest new therapeutic strategies for neuroinflammatory disorders.
Keywords: brain, chemokine, cytokine receptor, delayed hypersensitivity, experimental allergic encephalomyelitis, protein structure function, receptor expression, Mycobacterium bovis, Poland, T lymphocyte, cell migration, cooperative study, disease /disorder model, inflammation, ligand, messenger RNA, model design /development, neuroregulation, pathologic process, RNase protection assay, enzyme linked immunosorbent assay, flow cytometry, immunocytochemistry, injection /infusion, laboratory mouse, neutralizing antibody, tissue /cell culture
Project start date: 2002-08-01
Project end date: 2006-05-31
7R03TW006012-03 (2004): $48483
5R03TW006012-02 (2003): $46400
CORTICAL DEMYELINATION EARLY IN MS
Richard M Ransohoff, Staff
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Abstract: This project integrates ongoing P50-supported research progress in the Ransohoff lab, and related work from the lab of our collaborator Claudia Lucchinetti, now serving as co-PI. The P50 competing renewal represents a substantial extension of work ongoing in the Pis´ labs. The emphasis is on cortical demyelination and meningeal inflammation early in MS, as well as leukocyte trafficking to the cortex and subarachnoid space. P50 research will be leveraged through Dr. Lucchinetti´s R01NS49577, on which Dr. Ransohoff is co-PI. During analysis of biopsy material from eariy MS cases in the Lucchinetti lab, cortical demyelination (CDM) proved to be a common finding and collaboration with the Ransohoff group disclosed frequent association with meningeal inflammation. Notably, biopsies were performed to examine white matter changes on MRI, and the findings of CDM with meningeal inflammafion were unexpected. These findings however correlate well with MRI reports demonstrafing diffuse and focal cortical damage in eariy MS. Taken in the context of other recent results, our data in early MS suggest the novel hypothesis that meningeal inflammation and CDM promote white matter lesions during the MS disease process. This hypothesis will be addressed in Specific Aims 1 and 2 and will include detailed phenotyping of leukocytes within CDM lesions, including analysis of chemokine receptors. Importanfiy, recent studies in animal models identified specific chemokine receptors as candidate trafficking determinants for meninges and cortex. In Aim 3, we will use a novel flow-based in-vitro blood-brain barrier (BBB) model to define which chemokine receptors are essential for transmigrafion, enabling the identificafion of potenfial therapeufic targets for selective suppression of meningeal inflammafion and cortical demyelinafion, and to refine our interpretation of research results obtained during performance of Aims 1 and 2
Keywords: ATGN; Address; Animal Model; Animal Models and Related Studies; Antigenic Determinants; Antigens; Apoptosis; Apoptosis Pathway; B blood cells; B-Cells; B-Lymphocytes; BLR1; BLR1 gene; BN-1 Gene; BONZO; Binding Determinants; Biopsy; Blood - brain barrier anatomy; Blood leukocyte; Blood monocyte; Blood-Brain Barrier; Body Tissues; Bp50; Bursa-Dependent Lymphocytes; Bursa-Equivalent Lymphocyte; C-C CKR-5 Gene; C-C CKR-6 Gene; C-C Chemokine Receptor Type 5 Gene; C-C Chemokine Receptor Type 6 Gene; CC-CKR-5 Gene; CC-CKR-6 Gene; CCCKR5 Gene; CCR-5 Gene; CCR-6 Gene; CCR5; CCR5 gene; CCR6; CCR6 gene; CD183; CD195 Antigen Gene; CD40; CDW40; CHEMR13 Gene; CKR-5 Gene; CKR-L2; CKR-L3 Gene; CKR5 Gene; CKR6 Gene; CKRL3 Gene; CMKAR3; CMKBR5 Gene; CMKBR6 Gene; CXCR-5; CXCR3; CXCR3 gene; CXCR5; CXCR6; CXCR6 gene; Cell Death, Programmed; Cells; Cervical Lymph Node; Cervical lymph node group; Chemokine (C-C) Receptor 5 Gene; Chemokine (C-C) Receptor 6 Gene; Chemokine Receptor-Like 3 Gene; Chronic; Collaborations; Competence; Cytokines, Chemotactic; DCR2 Gene; DRY-6 Gene; DRY6 Gene; Data; Demyelinations; Diffuse; Disease; Disorder; Elements; Epitopes; Event; G Protein-Coupled Receptor 29 Gene; GPR-CY4 Gene; GPR29 Gene; GPR9; GPRCY4 Gene; HIV-1 Fusion Co-Receptor Gene; Hemato-Encephalic Barrier; Homologous Chemotactic Cytokines; INFLM; IP10; IP10-R; In Vitro; Infiltration; Inflammation; Inflammatory; Injury; Intercrines; LARC Receptor Gene; Lesion; Leukocyte Trafficking; Leukocytes; Lymph node proper; Lymphoid Follicle; MDR15; MGC9013; MHC Receptor; MR Imaging; MR Tomography; MRI; Magnetic Resonance Imaging; Magnetic Resonance Imaging Scan; Major Histocompatibility Complex Receptor; Marrow leukocyte; Marrow monocyte; Mediating; Mediator; Mediator of Activation; Mediator of activation protein; Medical Imaging, Magnetic Resonance / Nuclear Magnetic Resonance; Memory; Meningeal; Meninges; Mig-R; MigR; Modeling; Mononuclear; Myelin; NMR Imaging; NMR Tomography; Nuclear Magnetic Resonance Imaging; Pathogenesis; Pathology; Pathway interactions; Performance; Phenotype; Process; Publishing; Receptors, Antigen, T-Cell; Regulation; Reporting; Research; Research Support; Reticuloendothelial System, Leukocytes; Reticuloendothelial System, Lymph Node; Role; SIS cytokines; STRL22 Gene; STRL33; Seven-Transmembrane Receptor, Lymphocyte, 22 Gene; Subarachnoid Space; T memory cell; T-Cell Receptor; T-Cells; T-Lymphocyte; TNFRSF5; TNFRSF5 gene; TYMSTR; Thymus-Dependent Lymphocytes; Tissues; Tumor Necrosis Factor Receptor Superfamily Member 5 Gene; White Blood Cells; White Cell; Work; Zeugmatography; base; chemoattractant cytokine; chemokine; chemokine receptor; disease/disorder; gray matter; immunogen; in vitro Model; lymph gland; lymph nodes; macrophage; memory T lymphocyte; model organism; monocyte; novel; p50; pathway; social role; substantia alba; substantia grisea; therapeutic target; thymus derived lymphocyte; trafficking; white blood cell; white blood corpuscle; white matter; white matter change
2P50NS038667-11_5116 (2010): $142479
Sponsored Links Excellgen http://Excellgen.com
Chemokines And Chemokine Receptors In Multiple Sclerosis
Richard M Ransohoff, Director
Ccl College Of Med Of Cwru
Grant 5P50NS038667-090001 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Abstract: The overall goal of this project is to gain insight into cell trafficking and activation in multiple sclerosis (MS). We focus on chemokines and chemokine receptors, which are expressed both on infiltrating leukocytes and microglia in the central nervous system (CNS). Our general hypothesis is that chemokines and their receptors help determine the specific character of inflammatory CNS infiltrates. Recently, successful clinical trials of anti-leukointegrin antibodies validated the approach of blocking leukocyte trafficking to suppress inflammation in MS and phase II trials of small molecule chemokine receptor antagonists for treating MS are in progress. Therefore, it is increasingly important to delineate how chemokines and their receptors function during the evolution of MS, and clarify appropriate treatment targets. The Specific Aims of this competing renewal will focus on the following key issues First, how do chemokines regulate initial entry of blood-borne leukocytes into the CSF and CNS parenchyma? Second, how do chemokines govern migration and activation of blood-derived leukocytes and microglia within the inflamed parenchyma ? Third, what is the role of chemokines and their receptors in the compartmentalized inflammatory reaction of progressive MS? Our proposed research will examine in detail the roles of chemokines and their receptors in the pathogenesis of MS in its varied forms and phases, and will identify targets for treatment with chemokine receptor antagonists
Keywords: cell migration, chemokine, chemokine receptor, leukocyte, leukocyte activation /transformation, multiple sclerosis, pathologic process T lymphocyte, astrocyte, blood brain barrier, inflammation, microglia, monocyte, receptor expression clinical research, human tissue
MOLECULAR BASIS OF INTERFERON ACTION--RELATION TO M.S.
Richard M Ransohoff, Director
Cleveland Clinic Foundation 9500 Euclid Ave. Cleveland, Oh 44195
Grant 5K08NS001265-06 from National Institute Of Neurological Disorders And Stroke IRG: NSPA
Abstract: The long term goal of this proposal is to provide with a solid base in molecular biology. This foundation will enable him to address fundamental issues in neuroimmunology. The candidate will meet these objectives by continuing his involvement in determining molecular mechanisms of gene regulation by interfereons. This work, ongoing for three years, has consisted in two complementary projects. In the first, we investigated the mechanisms by which influenza virus replication is inhibited in interferon-treated cells. We found that viral mRNA transcription was abolished in MDBK cells treated with human alpha interferon. This observation led to further experiments using nuclear extracts of influenzainfected HeLa cells as in in vitro viral transcription system. Requiring a source of purified interferon-induced proteins, we constructed a cDNA library from interferon-treated MDBK cells in lambda GT 11. This library will be screened for clones representing interferon-induced mRNAs. Clones will be usd in hybrid selection/cell-free translation/in vitro viral transcription experiments, to detect clones encoding proteins which mediate inhibition of influenza transcription. These experiments will lead directly to characterization of the interferon-induced activity which results in imparied influenza virus transcription. The second project concerns the molecular regulation of expression of Ia (immune-associated) genes in human astorcytes. We have identified an astocytic tumor line, in which constitutive Ia expression is markedly enhanced by recombinant gamma interferon. Preliminary experiments indicate that this effect is specific for gamma interferon, and that mRNAs encoding Ia polypeptides accumulate to high levels in a dose-dependent, co- ordinate fashion ininterferon-treated cells. In these astrocytic cells and in a monocytic cell line, U 937, we will characterize kinetics of induction of Ia-encoding mRNAs and determine whether accumulation of these mRNAs is under transcriptional control. We will then clarify whether Ia expression on human astrocytes can e inhibited by other interferons or prostaglandins. These experiments will address issues of fundamental and clinical relevance in intrathecal immune responses.
Keywords: astrocyte, gene expression, interferon, multiple sclerosis, neuroimmunomodulation, complementary DNA, genetic library, genetic regulation, genetic transcription, genetic translation, messenger RNA, nucleic acid sequence, prostaglandin, human tissue, molecular cloning, molecular pathology, tissue /cell culture
Project start date: 1988-03-01
Project end date: 1993-02-28
5K08NS001265-06 (1992): $78300
CNS CHEMOKINE RECEPTOR EXPRESSION DURING CH R EAE
Richard M Ransohoff, Director
Cleveland Clinic Foundation
9500 Euclid Ave.
cleveland, Oh 44195
Grant 5R03TW000784-03 from Fogarty International Center IRG: ICP
Abstract: The current application is based on the hypothesis that CNS cells, including those of non-hematopoietic lineage, expressed chemokine receptors (CKRs) and thereby respond to local chemokine production. The proposal intends to define the level and cellular sources of CKR expression in the resting and inflamed CNS. It is virtually certain that infiltrating leukocytes will express the appropriate CKRs during chronic relapsing EAE. It remains unknown whether CKR expression on endogenous neural cells is upregulated during inflammation. The hypothesis will be addressed with three specific aims (1) analysis of steady state levels of CKR, mRNA expression in the resting CNS and during Ch-R EAE; (2) identification of cells that express CKRs within the resting CNS and during Ch-R EAE; (3) analysis of CKR expression following stereotactic injection of TNF-a injection
Keywords: central nervous system, chemokine, cytokine receptor, experimental allergic encephalomyelitis, receptor expression brain cell, inflammation, leukocyte, messenger RNA, neuron, tumor necrosis factor alpha Animalia, immunocytochemistry, polymerase chain reaction, tissue /cell culture
Project start date: 1997-09-01
Project end date: 2001-08-31
5R03TW000784-03 (1999): $20000
5R03TW000784-02 (1998): $20000
Chemokines In CNS Inflammation
Richard M Ransohoff, Director
Neurosciencescleveland Clinic Lerner Col/med-cwru
Grant 2R01NS032151-14A1 from National Institute Of Neurological Disorders And Stroke IRG: CNBT
Abstract: The current application focuses on CXCR2, a chemokine receptor present both on leukocytes and parenchymal central nervous system (CNS) cells. Our preliminary findings and recent reports demonstrate that CXCR2 on circulating leukocytes is decisive with regard to the outcome of EAE or cuprizone-mediated demyelination. Our recent findings show further that CXCR2 on CNS cells also regulates myelin repair. CXCR2 is the first chemokine receptor which promotes myelin destruction and hinders myelin repair. Our studies will address the fundamental role of CXCR2 in demyelination and remyelination, by performance of the following Specific Aims Aim 1. We will define the pathways by CXCR2 expression by leukocytes contributes to cuprizone- mediated demyelination. Hypothesis CXCR2 promotes demyelination through recruitment of monocytes to cuprizone lesions. The vast majority of hematogenous inflammatory cells in cuprizone lesions are monocyte- derived macrophages, leading us to suggest that CXCR2 acts through macrophage recruitment. The results of our proposed experiments will define how myeloid-cell CXCR2 contributes to this process. Aim 2. We will use an in-vitro model of demyelination-remyelination to address how the presence of CXCR2 impairs remyelination. Preliminary results showed that antibody-mediated blockade or genetic absence of CXCR2 led to accelerated remyelination of brain slices in-vitro. Hypothesis The presence of CXCR2 on oligodendrocyte progenitor cells impairs their proliferation and differentiation in demyelinated lesions. This in-vitro model recapitulates remyelination in-vivo, particularly those aspects which take place within the demyelinated lesion. This convenient, manipulable model will be used to examine the cellular and molecular basis by which CXCR2 hinders remyelination. Aim 3. We will define CNS cells whose expression of CXCR2 leads to inefficient remyelination. To address CXCR2 function in the CNS, we generated a conditional CXCR2 allele. Hypothesis CXCR2 impairs remyelination by slowing accumulation of oligodendrocyte progenitors in lesions and hindering their proliferation and differentiation. Mice harboring a conditional CXCR2 allele will be crossed with available mice expressing cre-ER(T2) in individual cell types, allowing efficient lineage-specific, inducible deletion. We will analyze cuprizone-mediated demyelination and remyelination to probe how CXCR2 expression in CNS cells regulates this process. These studies will provide essential data for designing clinical trials of CXCR2 blockade in MS. This grant focuses on CXCR2, a cellular receptor present both on inflammatory white blood cells and brain cells. CXCR2 both promotes myelin destruction and hinders myelin repair. Our studies will address the fundamental role of CXCR2 in demyelination and remyelination, by using novel models including in-vitro brain slices and genetically modified mice, allowing the receptor to be removed either from inflammatory white blood cells or from CNS cells. Ongoing clinical trials for pulmonary disease use drugs that block CXCR2 and our studies will provide essential data for designing clinical trials of CXCR2 blockade in MS
Project start date: 1994-05-01
Project end date: 2014-01-31
Core--Tissue Acquistion/characterization And Biostatistics
Richard M Ransohoff, Director
Cleveland Clinic Foundation 9500 Euclid Ave. Cleveland, Oh 44195
Grant 5P01NS038667-059001 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Abstract: SUBPROJECT NOT AVAILABLE
Keywords: biomedical facility, central nervous system, histopathology, multiple sclerosis, neuropathology, statistics /biometry, tissue donor, human tissue
Sponsored Links Excellgen http://Excellgen.com
Tissue Injury And Inflammation In Multiple Sclerosis
Richard M Ransohoff, Director
Molecular Medicinecleveland Clinic Lerner Col/med-cwru
Grant 5P50NS038667-10 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Abstract: Multiple sclerosis (MS) remains the leading cause of neurological nontraumatic disability for young adults in North America. This Program Project competing renewal application is a direct extension of our research during the past period of support, and is based on interlinked hypotheses regarding the inflammatory destructive and neurodegenerative processes in MS patients, which begins from disease onset and culminates in progressive neurologic disability due to accumulated, irreversible tissue injury. The program consists of four related projects and one multifunctional core. Project 1 Chemokines and chemokine receptors in multiple sclerosis. The competing renewal will define in detail how individual receptors govern infiltration of monocytes in acute lesions and address for the first time differential trafficking by CD4+ and CD8+ T cells. We intend further to determine how chemokine receptors govern movement of T cells and APCs in acute and chronic MS lesions. We will use a BBB model to address the differences we observe in chemokine receptor expression by cells in blood, CSF and brain parenchyma. Proiect 2 Axonal pathology in multiple sclerosis proposes that axonal pathology is a primary contributor to neurological deficits in MS patients and should be considered as a therapeutic target. The competing renewal focuses on molecular mechanisms of neurodegeneration in MS brain and spinal cord, based upon the hypothesis that degeneration of chronically-demyelinated axons results from reduced axonal energy metabolism, which, in turn, alters the ionic milieu of the axoplasm, culminating in increased intraxonal Ca++ and axonal degeneration. Project 3 Monitoring brain atrophy during the course of multiple sclerosis has established that brain atrophy is a practical, sensitive and relevant surrogate marker of the underlying disease process. However, determinants of variance in rate of atrophy remain undefined. This issue will be addressed by use of advanced MRI techniques to analyze changes in individual lesions and examination of diffuse and regional tissue damage. Proiect 4 Biomarkers of the therapeutic response in MS will use innovative strategies for determining IFN responder status and cDNA macroarrays to monitor the genetic program of response to IFN, to address molecular markers of response to IFN in MS patients. Core Tissue acquisition, biostatistics, MRI imaging, administration will provide data analysis/management and administrative support for projects, establish, maintain and distribute for projects a unique resource of MS autopsy tissue, and coordinate imaging protocols. This research program will continue to impact our understanding of MS and lead to novel treatments
Keywords: multiple sclerosis, pathologic process clinical research
Project start date: 1999-12-01
Project end date: 2009-11-30
5P01NS038667-08 (2007): $1184550
5P01NS038667-07 (2006): $1198234
2P01NS038667-06 (2005): $1264874
TISSUE INJURY AND INFLAMMATION IN MS (P50)
Richard M Ransohoff, Staff
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Grant 2P50NS038667-11 from National Institute Of Neurological Disorders And Stroke
Abstract: Multiple sclerosis (MS) remains the leading cause of neurological nontraumatic disability for young adults in North America and outcomes of current therapy are often unsatisfactory. This P50 competing renewal application directly extends and amplifies our research during the previous two cycles of support. We focus here on gray matter pathology, extending our previous work addressing interlinked hypotheses regarding the destructive inflammatory and neurodegenerative processes in MS patients. The P50 encompasses several unique resources including a rapid-autopsy program which has yielded insights into MRI-pathology correlations; a longitudinal MRI cohort which has illuminated the mechanisms underlying brain atrophy in MS; and newly-recruited, a series of MS biopsy cases which can be studied to understand cortical pathology early in MS. Core A Tissue acquisition, imaging, biostatistics. administration (R Ransohoff) will establish, maintain and distribute a unique resource of MS autopsy tissue with dedicated postmortem imaging, as well as coordinate database management and provide administrative and biostatistical support for all projects. Project 1 Cortical demyelination and leukocyte trafficking early in MS (R Ransohoff; C Lucchinetti) will address the hypothesis that meningeal inflammation and cortical demyelination early in the MS process are implicated in facilitating white matter demyelination. This project will also address mechanisms of selective leukocyte trafficking to meninges and cortex. Project 2 Cellular and molecular mechanisms of cortical remyelination in MS patients (B Trapp) will examine cortical demyelination and remyelination in MS brains, and characterize both the cells mediating repair and the factors that inhibit repair. Project 3 Gray matter atrophy in multiple sclerosis (E Fisher) will use novel quantification techniques, developed as part of the P50 research, address in a longitudinal MRI cohort, the relationship between gray matter atrophy and white matter lesions as well as normal-appearing white matter changes. This project will also define relationships between gray matter atrophy and neuropsychological changes. Project 4 Gray matter atrophy in multiple sclerosis Clinical implications (R Rudick) will extend the use of our new MRI analytic software to define the time course of gray matter atrophy, and evaluate the effects of immunomodulatory therapy, using files from completed clinical trials of a diversity of agents Extending our established research approaches to address gray matter pathology in MS will provide new insights into disease mechanisms and identify novel therapeutic targets. Although MS has been considered a "white matter disease", it also affects the gray matter. All four projects that comprise this P50 focus on gray matter in MS. The P50 includes studies to understand the pathologic and repair mechanisms in gray matter, to develop new imaging markers to measure gray matter pathology, and to characterize gray matter pathology over the course of MS. Ultimately, this research will lead to new therapeutic targets and new ways to monitor neural protection to improve the lives of MS patients. Disclaimer Please note that the following critiques were prepared by the reviewers prior to the Study Section meeting and are provided in an essentially unedited form. While there is opportunity for the reviewers to update or revise their written evaluation, based upon the group´s discussion, there is no guarantee that individual critiques have been updated subsequent to the discussion at the meeting. Therefore, the critiques may not fully reflect the final opinions of the individual reviewers at the close of group discussion or the final majority opinion of the group. Thus the Resume and Summary of Discussion is the final word on what the reviewers actually considered critical at the meeting. OVERALL PROGRAM EVALUATION
Keywords: No Project Terms available
Project start date: 1999-12-01
Project end date: 2014-12-31
Budget start date: 18-JAN-2010
Budget end date: 31-DEC-2010
2P50NS038667-11 (2010): $1048242
TISSUE INJURY AND INFLAMMATION IN MULTIPLE SCLEROSIS
Richard M Ransohoff, Director
Cleveland Clinic Foundation 9500 Euclid Ave. Cleveland, Oh 44195
Grant 5P01NS038667-05 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Abstract: Multiple sclerosis (MS) is the leading cause of neurological non- traumatic disability for young adults in Norther America. This revised Program Project application is based on the hypothesis that the inflammatory destructive process in MS patients is active from disease onset and that progressive neurologic disability is a late complication of accumulated, irreversible tissue injury. The program consists of three related projects and one core. Project 1 Chemokines and chemokine receptors in multiple sclerosis (RM Ransohoff, PI) will address the hypothesis that specific chemokines and receptors are significantly involved in central nervous system (CNS) inflammation during MS. To define molecular targets for therapy, we will focus on chemokines and receptors expressed by T cells and mononuclear phagocytes in MS. PROJECT 2 Axonal pathology in multiple sclerosis (BD Trapp) is based on the hypothesis that axonal pathology is a primary contributor to neurological deficits in MS patients and should be considered as a therapeutic target. Axonal pathology will be characterized in spinal cords of patients dying from MS; in rats subjected to spinal cord transection and in mice with experimental autoimmune encephalomyelitis (EAE). Data will be correlated with biochemical measures of N-acetyl aspartate of N-acetyl aspartate, a reflection of axonal injury. In aim 4, oligodendrocyte progenitors in MS brain will be characterized, because chronic deprivation of the trophic support of myelin is responsible in part for anatomic interruption of axons in MS lesions. Project 3 Monitoring brain atrophy during the course of multiple sclerosis (RA Rudick) will test the hypothesis that brain atrophy will be a practical, sensitive and relevant surrogate marker of the underlying disease process. Recent studies indicate that brain atrophy can be measured from the early stages of MS in relapsing-remitting patients. Project 3 will address the rate and temporal pattern of brain atrophy in MS patients, and will establish relationships between brain atrophy and clinical disability in MS patients. The relation between brain atrophy on MRI and various additional MRI lesion types will be established, and we will perform MR/pathological correlations from scans performed at the time of autopsy during our tissue acquisition protocol. Core Tissue acquisition, biostatistics, administration (RA Rudick) will establish, maintain and distribute for projects a unique resource of MS autopsy tissue, histological and post- mortem imaging data. This core will provide data analysis/management and administrative support for projects. This research program will directly impact monitoring and treatment of MS.
Keywords: inflammation, multiple sclerosis
Project start date: 1999-12-01
Project end date: 2004-11-30
5P01NS038667-05 (2004): $846517
5P01NS038667-04 (2003): $939818
5P01NS038667-03 (2002): $925059
5P01NS038667-02 (2001): $931980
Sponsored Links Excellgen http://Excellgen.com
1P01NS038667-01A1 (2000): $853803
GENDER DIFFERENCES IN IMMUNE RESPONSES IN MS
Richard M Ransohoff, Staff
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Grant 5R01NS041972-06 from National Institute Of Neurological Disorders And Stroke
Abstract: Most autoimmune diseases occur with higher frequency in women. This suggests that the autoimmune phenotype is related, at least in part, to immunologic effects from sex hormones. To date, most in vitro and in vivo studies have shown that high, pregnancy doses of estrogen and progesterone are immune suppressive. Unfortunately, these data do not explain why more women get multiple sclerosis (MS) compared to men. Evidence showing a Th1 -immune promoting effect by estrogen is minimal, although such data would help explain the gender bias in susceptibility to autoimmune disease. In the previous funding period, we presented evidence to support gender effects in the cytokine response to myelin women with MS show dramatically skewed IFN? responses to certain myelin epitopes, whereas men with MS show an IL-5 response. In this proposal, we present evidence that 1) endogenous sex hormone levels are associated with gender-specific cytokine responses; 2) the strength of the T cell receptor stimulus, along with hormone dose, may determine whether the resulting immune response is enhanced or suppressed; and 3) low doses of estrogen promote memory CD4 T cell migration across brain endothelium. These data have lead us to hypothesize that normal ovarian cycle doses of estrogen regulate four important steps in the pathogenesis of MS 1) promote Th1 cellular activation; 2) promote lymphocyte adhesion to brain endothelium; 3) regulate chemoattraction of lymphocytes to endothelium; and 4) facilitate migration of memory CD4 T cells across the blood brain barrier (BBB). This proposal will address this hypothesis by studying the migration of human peripheral blood lymphocytes across a well-characterized migration model of the BBB consisting of human brain microvascular endothelial cells. In summary, this study will define the role of estrogen in regulating cell activation via cytokine secretion and proliferation; adhesion molecule expression and function on lymphocytes and on brain endothelial cells; attraction of lymphocytes across brain endothelium via chemokines and chemokine receptors; and invasion of the BBB via matrix metalloproteinases and migration of pathogenic lymphocytes across the BBB. These studies will provide valuable understanding of the multiple effects of estrogens and may provide molecular targets for development of future therapies as well as a better understanding of the role of sex hormones in the pathogenesis of MS
Keywords: Address; Adhesion Molecule; Adhesions; Affect; Anti-Inflammatories; Anti-Inflammatory Agents; Anti-inflammatory; Antigenic Determinants; Antiinflammatories; Antiinflammatory Agents; Aquadiol; Autoimmune; Autoimmune Diseases; Autoimmune Process; B Cell Differentiation Factor I; B cell growth factor 2; B-Cell Growth Factor-II; BCGF-II; BCGF2; BCGF2 (B cell growth factor 2); Binding Determinants; Blood - brain barrier anatomy; Blood leukocyte; Blood-Brain Barrier; Brain; Cell Adhesion Molecules; Cell Growth in Number; Cell Locomotion; Cell Migration; Cell Movement; Cell Multiplication; Cell Proliferation; Cells; Cellular Migration; Cellular Proliferation; Chemotherapy-Hormones/Steroids; Corpus Luteum Hormone; Cytokines, Chemotactic; Data; Delta4-pregnene-3, 20-dione; Development; Dimenformon; Diogyn; Diogynets; Doctor of Philosophy; Dose; EDF; Encephalon; Encephalons; Endocrine Gland Secretion; Endothelial Cells; Endothelium; Eo-CSF; Eosinophil Differentiation Factor; Epitopes; Estra-1, 3, 5(10)-triene-3, 17-diol (17beta)-; Estrace; Estradiol; Estradiol-17 beta; Estradiol-17beta; Estraldine; Estrogen Receptors; Estrogenic Agents; Estrogenic Compounds; Estrogens; Frequencies (time pattern); Frequency; Funding; Future; Gender; Gender Bias; Gestation; Gonadal Steroid Hormones; Hemato-Encephalic Barrier; Homologous Chemotactic Cytokines; Hormones; Human; Human Migration / Distribution; Human, General; IFN; IL-5; IL5; IgA enhancing factor; Immigrations; Immune; Immune response; Immunologic, Immunochemical; Immunologics; In Vitro; In-Migration; Inflammatory; Inflammatory Response; Intercrines; Interferons; Interleukin 5 (Colony-Stimulating Factor, Eosinophil); Interleukin 5 Precursor; Interleukin-5; Investigators; Lead; Leukocytes; Lymphocyte; Lymphocytic; MHC Receptor; MMPs; MS (Multiple Sclerosis); Major Histocompatibility Complex Receptor; Man (Taxonomy); Man, Modern; Marrow leukocyte; Matrix Metalloproteinases; Memory; Modeling; Molecular Target; Motility; Motility, Cellular; Multiple Sclerosis; Myelin; Nervous System, Brain; Ovarian Cycles; Ovocyclin; Ovocylin; PBL; PBMC; Pathogenesis; Patients; Pb element; Peripheral Blood Lymphocyte; Peripheral Blood Mononuclear Cell; Ph.D.; PhD; Phenotype; Play; Predisposition; Pregn-4-ene-3, 20-dione; Pregnancy; Pregnenedione; Progesterone; Programs (PT); Programs [Publication Type]; Progynon; Receptors, Antigen, T-Cell; Regulation; Research Personnel; Researchers; Rest; Reticuloendothelial System, Leukocytes; Role; SIS cytokines; Sclerosis, Disseminated; Sex Bias; Sex Characteristics; Sex Differences; Sex Hormones; Sex Steroid Hormones; Sexism; Specificity; Stimulus; Susceptibility; T cell replacing factor; T-Cell Receptor; T-Cell Replacing Factor; Testing; Therapeutic Estradiol; Therapeutic Estrogen; Therapeutic Hormone; Therapeutic Progesterone; White Blood Cells; White Cell; Woman; autoimmune disorder; cell adhesion protein; cell motility; chemoattractant cytokine; chemokine; chemokine receptor; cytokine; gender difference; gonadal steroids; heavy metal Pb; heavy metal lead; host response; human migration; immunoresponse; in vivo; insular sclerosis; lymph cell; memory CD4 T cell; memory CD4 T lymphocyte; men; men`s; migration; programs; response; sex steroid; sexual dimorphism (noncellular); social role; white blood cell; white blood corpuscle
Project start date: 2001-08-01
Project end date: 2010-06-30
Budget start date: 1-JUL-2009
Budget end date: 30-JUN-2010
5R01NS041972-06 (2009): $270375
5R01NS041972-05 (2008): $270375
2R01NS041972-04A2 (2007): $270375
ASTROCYTE SYNTHESIS OF CHEMOTACTIC CYTOKINES IN EAE
Richard M Ransohoff, Director
Cleveland Clinic Foundation
9500 Euclid Ave.
cleveland, Oh 44195
Grant 1R01NS032151-01A1 from National Institute Of Neurological Disorders And Stroke IRG: NEUC
Abstract: Inflammatory cell recruitment to the central nervous system (CNS) is a cardinal feature of numerous pathological processes, including multiple sclerosis (MS). Despite recent progress, the soluble signals that attract inflammatory cells from perivascular accumulations into the CNS parenchyma remain obscure. The present proposal derives from experience in our laboratories and is based on the hypothesis that a specific class of chemoattractant cytokines termed ´chemokines´ is synthesized by astrocytes in response to inflammatory stimuli; we propose that astrocyte-derived chemokines in turn function to recruit inflammatory cells into the CNS parenchyma. This hypothesis will be addressed by performance of two Specific Aims Al. We will establish the relationship between chemokine gene expression and CNS inflammation in informative model systems. Three models will be studied, each with specific advantages 1) EAE of SJL/J mice actively immunized with proteolipid protein (PLP) encephalitogens; 2) EAE of Lewis rats, after adoptive transfer of myelin basic protein (MBP)-specific or myelin oligodendroglial glycoprotein (MOG)-specific T-cell lines; 3) Traumatic brain injury in neonatal and adult mice. We will assess chemokine transcript levels by a sensitive, semi-quantitative reverse transcription/polymerase chain reaction (RT/PCR) assay. Subsequently, we will establish the relationship between specific histopathologic alterations and localized chemokine production in CNS tissues, assayed by in situ hybridization (ISH). A2. We will evaluate regulation of chemokine gene expression in astrocytic cells in vitro and murine CNS in vivo. Our preliminary work showed that CNS expression of chemokine mRNAs was limited to astrocytes in mice with EAE. To determine the mechanism of this chemokine gene expression, we will address the cytokine regulators of IP-10 and JE/MCP-l transcription in human astrocytic and monocytic cells in vitro and in murine CNS cells in vivo 1) Regulation of chemokine gene expression in human astrocytic and monocytic cells will be compared. 2) Chemokine gene expression in the murine CNS will be determined by RT/PCR and ISH in vivo after intravenous injection of pro-inflammatory cytokines IFNgamma and TNFalpha. The production of chemoattractant cytokines by astrocytes provides insight into the role of endogenous neural cells in amplifying inflammatory processes. Research proposed here will enhance our understanding of varied pathologic processes and suggest potential therapeutic strategies for disorders limited to the CNS
Keywords: astrocyte, experimental allergic encephalomyelitis, interferon gamma, neuroimmunomodulation, tumor necrosis factor alpha brain injury, chemoattractant, gene deletion mutation, gene induction /repression, genetic promoter element, inflammation, monocyte, myelin basic protein, myelin glycoprotein, myelin proteolipid, protein biosynthesis, transcription factor, trauma human tissue, in situ hybridization, laboratory mouse, laboratory rat, newborn animal, polymerase chain reaction, tissue /cell culture
Project start date: 1994-05-01
Project end date: 1998-04-30
1R01NS032151-01A1 (1994): $133079
Chemokines In CNS Inflammation
Richard M Ransohoff, Director
Cleveland Clinic Lerner Col/med-cwru P84 Cleveland, Oh 44195
Grant 3R01NS032151-12S1 from National Institute Of Neurological Disorders And Stroke IRG: ZRG1
Abstract: Chemokines and their G-protein coupled receptors constitute a unique ligand-receptor system that governs leukocyte migration and effector function. Chemokines are present in most multicellular organisms, to mediate organ patterning during development. In mammals, chemokines also regulate the function of a sophisticated, flexible host-defense system. The central nervous system (CNS) lesions of multiple sclerosis (MS), an inflammatory demyelinating disease, contain specific populations of activated leukocytes, including monocytes, macrophages, microglia and lymphocytes. Understanding the molecular determinants of leukocyte trafficking to the CNS will promote progress towards effective treatment of MS. Data obtained in the previous round of funding supported our core hypothesis that individual chemokine/receptor pairs are selectively involved in the recruitment and activation of specific leukocyte populations in the CNS. Using experimental autoimmune encephalomyelitis (EAE) as a model system, we propose to examine our hypothesis in detail. We have carefully selected genetically-modified mice that will allow us to examine chemokine signaling in the critical cell populations implicated in EAE and MS. Using these powerful reagents, we intend to clarify how chemokines and their receptors govern the CNS recruitment and activation of diverse cell populations that take part in EAE monocytes, lymphocytes, astrocytes, NK cells and microglia. The Specific Aims address the following questions 1. How do MCP- l/CCL2 and CCR2 govern cell recruitment and activation in the CNS? 2. How do fractalkine/CX3CL1 and CX3CR1 regulate the recruitment and activation of resident microglia and infiltrating cells in the CNS of mice with EAE? 3. How do CXCR3 and its ligands govern the accumulation and distribution of T-cells in the CNS of mice with EAE?
Keywords: astrocyte, cell migration, chemokine, cytokine receptor, experimental allergic encephalomyelitis, gene expression, lymphocyte, microglia, monocyte, natural killer cell, receptor expression, IP 10 protein, cell motility, cytotoxic T lymphocyte, helper T lymphocyte, macrophage inflammatory protein, monocyte chemoattractant protein 1, cell line, genetically modified animal, immunocytochemistry, laboratory mouse, polymerase chain reaction, tissue /cell culture
Project start date: 1994-05-01
Project end date: 2007-03-31
3R01NS032151-12S1 (2005): $20000
7R01NS032151-11 (2004): $363375
2R01NS032151-09 (2002): $351500
CYTOKINE CROSSTALK IN ASTROCYTOMA CELLS
Richard M Ransohoff, Director
Institution:
Grant 5P01CA062220-030003 from National Cancer Institute
Abstract: Astrocytoma is the most common primary neoplasm of the human central nervous system, and current therapy for this devastating disease is unsatisfactory. The overall objective of research proposed in this application is to investigate the molecular mechanism by which signaling interactions between interferon-gamma (IFN(gamma)) and transforming growth factor beta-1 (TGF(beta1)) occur in human astrocytoma cells. This approach was selected because TGF(beta1) and IFN(gamma) are present within glial tumor tissue and evoke biologically meaningful responses from astrocytoma cells. Detailed mechanistic understanding of interactions between counter-regulatory cytokines such as IFN(gamma) and TGF(beta1) may therefore be significant for elucidating the biology and treatment of glioma. We showed that TGF(beta1) inhibited expression of two IFN(gamma)- inducible genes, HLA-DRA and IDO, in a human astrocytoma cell line. The patterns of IFN(gamma)-inducible expressions and regulatory cis elements of these two genes were different, indicating that HLA-DRA and IDO can be used as distinct model systems to investigate crosstalk between TGF(beta1) and IFN(gamma). The specific Aims are 1. To investigate the mechanism by which IFN(gamma)-induced HLA-DRA transcription is impaired in TGF(beta1)-treated astrocytoma cells. We will extend our current studies by addressing protein factors binding to the HLA-DRA promoter in IFN(gamma)- and TGF(beta1)-treated cells. We will determine the cis-element needed for inhibiting transcription of the invariant chain (INV) gene, which is co-regulated with HLA-DRA, and evaluate TGF(beta1) effects on factor-binding to this element. Factors required for HLA-DRA transcription will be cloned, using a novel strategy based on retroviral expression of cDNAs encoding genetic suppressor elements (GSE) and immunoselection against HLA-DRA antigen expression. 2. To investigate TGF(beta1) inhibition of the IFN(gamma)-inducible IDO gene. We will use transfection studies to determine the cis-element required for TGF(beta1) inhibition of the 2,3-indoleamine dioxygenase (IDO) gene, and band-shifts to evaluate the effect of TGF(beta1) on factor-binding to these regulatory elements. Experiments proposed in this application will provide significant novel insight into signaling in astrocytoma cells by biologically important cytokines.
Keywords: astrocytoma, biological signal transduction, genetic regulation, genetic transcription, interferon gamma, transforming growth factor, DNA binding protein, gene induction /repression, genetic promoter element, genetic regulatory element, histocompatibility gene, neoplastic cell, oxygenase, protein biosynthesis, transcription factor, DNA footprinting, crosslink, flow cytometry, gel mobility shift assay, immunocytochemistry, molecular cloning, reporter gene, tissue /cell culture
Sponsored Links Excellgen http://Excellgen.com
CHEMOKINES IN CNS INFLAMMATION
Richard M Ransohoff, Director
Cleveland Clinic Foundation
9500 Euclid Ave.
cleveland, Oh 44195
Grant 3R01NS032151-06S1 from National Institute Of Neurological Disorders And Stroke IRG: ZRG1
Abstract: Applicant´s ) The long-term goal of this research is to define the roles of selected chemokines in central nervous system (CNS) inflammatory responses involving monocytes, macrophages and lymphocytes. Chemokines are small pleiotropic chemoattractant cytokines that mediate inflammatory, angiogenic and growth-regulatory functions. We propose to define the roles of two chemokines, monocyte chemoattractant protein-lalpha (MCP-1alpha) and macrophage inflammatory protein-lalpha (MiP-lalpha) in the induction of inflammation in the CNS. To achieve this goal, we will use the powerful and well-characterized models that are relevant for human disease, namely experimental autoimmune encephalomyelitis (EAE) and spinal cord contusion injury (SCI). We will determine molecular, histological and behavioral characteristics of the models in genetically altered mice that are ´knocked-out´ for MCP-1 or MIP-lalpha ligands or receptors, or that overexpress these chemokines in the CNS. The data gained in these studies will provide insight into the pivotal roles of chemokines and their target cells in injury responses of the CNS. It is essential to understand how these important chemokine mediators are expressed in relation to development of pathologic states. We will demonstrate how varied stimuli converge to regulate chemokine expression by CNS cells, using two incisive models of chemokine gene regulation Theiler´s murine encephalomyelitis virus (TMEV) infection of the CNS of wild-type and cytokine-knockout mice; and a defined tissue-culture system. The Specific Aims address 1. Role of CC chemokines in experimental autoimmune encephalomyelitis. 2. Regulation of chemokine expression during TMEV-induced inflammation. 3. MCP-1 and its receptor in spinal cord contusion injury. 4. Regulation of MCP-1 transcription by constitutive and inducible factors. Data gained from these experiments will provide a rational foundation for strategies to modulate chemokine expression during neurological disease states
Keywords: chemokine, experimental allergic encephalomyelitis, gene induction /repression, infectious encephalitis, spinal cord injury cytokine receptor, inflammation, microorganism immunology, monocyte chemoattractant protein 1, murine encephalomyelitis virus, neurogenetics laboratory mouse, tissue /cell culture, transgenic animal
Project start date: 1994-05-01
Project end date: 2002-05-31
3R01NS032151-06S1 (1999): $80000
2R01NS032151-05 (1998): $202646
Chemokines And Chemokine Receptors In Multiple Sclerosis
Richard M Ransohoff, Director
Cleveland Clinic Foundation 9500 Euclid Ave. Cleveland, Oh 44195
Grant 5P01NS038667-050001 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Abstract: adapted from page 125). The project will study chemokine receptors on T cells and mononuclear phagocytes (blood-borne monocytes, macrophages, microglia) in a set of related tissues Blood and CSF from MS patients, and blood and brains from marmosets with autoimmune disease. The stated hypothesis is that "chemokine receptors are significantly involved in leukocyte invasion, differentiation and activation in the CNS during MS." "Functional significance" will be studied by "quantitating receptor-bearing cells." It is expected that the work will "define molecular targets for therapy." (pg. 125, first par.) Aim 1 defines chemokine receptors on T cells in MS brain. The hypothesis is that T cells in MS lesions representing different stages of disease activity or patterns of demyelination will show different chemokine receptor profiles. The receptors to be studied are CXCR3 and CCR5, characteristic of Th1 cells; CCR4 and CCR3, characteristic of Th2 cells; and CXCR4, expressed by naive T cells. The PI will also ask which cells express IP-10 and RANTES, which are ligands for the Th1 receptors. Aim 2 performs a similar analysis for mononuclear phagocytes in MS brain, to test a similar hypothesis. The receptors to be studied are CCR2, characteristic of monocytes; CCR1, characteristic of macrophages; and CCR5, characteristic of phagocytic macrophages and activated microglia. Aim 3 defines the same receptors on cells from blood and CSF of MS patients (3a), and blood and brain from marmosets (3b). It is explained that AIM 3 "examines the functional significance of receptor expression (p. 125, last 3 lines)."
Keywords: chemokine, chemokine receptor, inflammation, multiple sclerosis, T lymphocyte, experimental allergic encephalomyelitis, macrophage, animal tissue, human tissue
Richard M Ransohoff
Cleveland Clinic Lerner Col/med-cwru
Project start date: 2011-03-01
Project end date: 2013-02-28
CHEMOKINE REGULATION ON CENTRAL NERVOUS SYSTEM INFLAMMATION IN MULTIPLE SCLEROSIS
Richard M Ransohoff, Staff
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Grant 5K24NS051400-05 from National Institute Of Neurological Disorders And Stroke
Keywords: Address; Anatomic; Anatomical Sciences; Anatomy; Antibodies; Attention; Biological Models; Blood - brain barrier anatomy; Blood leukocyte; Blood monocyte; Blood-Brain Barrier; Body Tissues; Brain; Cells; Central Nervous System; Characteristics; Clinical; Clinical Research; Clinical Study; Clinical Trials; Clinical Trials, Unspecified; Cytokines, Chemotactic; Data; Development; Disease; Disorder; Encephalon; Encephalons; Evolution; Familiarity; Generations; Goals; Hemato-Encephalic Barrier; Homologous Chemotactic Cytokines; Hortega cell; INFLM; Image; Inflammation; Inflammatory; Intercrines; Investigators; Lesion; Leukocyte Trafficking; Leukocytes; MR Imaging; MR Tomography; MRI; MS (Multiple Sclerosis); MS Lesions; Magnetic Resonance Imaging; Magnetic Resonance Imaging Scan; Marrow leukocyte; Marrow monocyte; Medical Imaging, Magnetic Resonance / Nuclear Magnetic Resonance; Mentors; Mentorship; Methodology, Research; Microglia; Model System; Models, Biologic; Multiple Sclerosis; Multiple Sclerosis Lesions; NMR Imaging; NMR Tomography; Nervous System, Brain; Nervous System, CNS; Neuraxis; Neurologic; Neurological; Nuclear Magnetic Resonance Imaging; Patients; Pattern; Phase; Programs (PT); Programs [Publication Type]; Receptor Protein; Regulation; Research; Research Methodology; Research Methods; Research Personnel; Research Technics; Researchers; Reticuloendothelial System, Leukocytes; SIS cytokines; Science of Anatomy; Scientist; Sclerosis, Disseminated; Series; Staging; T-Cells; T-Lymphocyte; Techniques, Research; Testing; Thymus-Dependent Lymphocytes; Tissues; United States; White Blood Cells; White Cell; Zeugmatography; anatomy; base; biomarker; chemoattractant cytokine; chemokine; chemokine receptor; clinical investigation; clinical practice; disability; disease/disorder; gitter cell; human disease; imaging; in vitro Model; in vivo; insight; insular sclerosis; mesoglia; microglial cell; microgliocyte; monocyte; natalizumab; novel; perivascular glial cell; programs; receptor; receptor expression; receptor function; selective expression; selectively expressed; small molecule; thymus derived lymphocyte; trafficking; white blood cell; white blood corpuscle
Project start date: 2006-02-08
Project end date: 2011-01-31
Budget start date: 1-FEB-2010
Budget end date: 31-JAN-2011
PFA/PA: PA-04-107
5K24NS051400-05 (2010): $169668
TISSUE ACQUISITION, IMAGING, BIOSTATISTICS, ADMINISTRATION CORE
Richard M Ransohoff, Staff
Cleveland Clinic Lerner Col/med-cwru, Jjn5-01, Cleveland, Oh 44195
Keywords: No Project Terms available
2P50NS038667-11_5120 (2010): $283158
Chemokine Regulation On Central Nervous System Inflammation In Multiple Sclerosis
Richard M Ransohoff, Director
Molecular Medicinecleveland Clinic Lerner Col/med-cwru
Grant 5K24NS051400-04 from National Institute Of Neurological Disorders And Stroke IRG: NST
Project start date: 2006-02-08
Project end date: 2011-01-31
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