The Role Of P38 MAP Kinase In IL-6 Gene Regulation
Keith L Kirkwood
State University Of New York At Buffalo Sponsored Projects Services Buffalo, Ny 14260
Grant 5R03DE014460-02 from National Institute Of Dental And Craniofacial Research IRG: DSR
Abstract: Chronic production of cytokines mediates inflammatory diseases. Proinflammatory cytokine production in periodontal disease, as well as other chronic inflammatory bone diseases, results in a net bone loss that ultimately negatively affects host function. A complex cytokine network controlled by many cell types, dictates cellular response in bone resorption. Proinflammatory cytokines, such as interleukin (IL)-6, regulate bone resorption through interaction with downstream cytokines. Intracellular signaling pathways regulate IL-6 through a variety of potential mechanisms. This current proposal will focus on the role of p38 mitogen activated kinase (MAP) and how this kinase regulates IL-6 mRNA in osteoblasts, p38 has been shown to play a significant role in regulation of mRNA stability of several inflammatory mediators including IL-6, IL-8, and COX-2. Post-transcriptional control of mRNA stability has been implicated as a potential role of p38 MAP kinase. The molecular mechanisms that underlie p38 regulation are poorly understood, but increasing evident from other studies suggests that cis elements of the 3 untranslated region (UTR) may play a major role. Preliminary data obtained for this proposal indicates that a specific p38 MAP kinase inhibitor, SB203580, can inhibit IL-l-induced expression of IL-6 in a dose dependent manner. The mechanism has been shown to depend upon de novo protein synthesis and occurs at the post-transcriptional level where IL-6 mRNA stability in dramatic decreased in the presence of SB203580. Thus, this proposal will focus establishing the role of p38 MAP kinase in controlling IL-6 regulation. The proposed studies may be important because they could lead towards the development of novel pharmacological agents that regulate IL-6 expression at the post-transcriptional level. The specific aims of the proposal are the following 1) To establish that p38 MAP kinase is a key regulator of IL-l-induced IL-6 production in osteoblastic cells through construction of dominate negative mutants of p38 MAP kinase and correlating p38 MAP kinase activity with IL-6 mRNA expression and stability, and 2) To determine the cis elements of IL-6 3 untranslated region (UTR) that mediate p38 MAP kinase-induced IL-6 mRNA stability. These studies will provide insight into how specific sequence elements of IL-6 are modulated by p38 MAP kinase and control mRNA decay rates.
Keywords: genetic regulation, interleukin 6, mitogen activated protein kinase, osteoblast, biological signal transduction, enzyme activity, enzyme mechanism, gene expression, interleukin 8, intracellular, messenger RNA, periodontitis, physiologic bone resorption, prostaglandin endoperoxide synthase, SDS polyacrylamide gel electrophoresis, enzyme linked immunosorbent assay, polymerase chain reaction, transfection /expression vector, western blotting
Project start date: 2002-08-01
Project end date: 2004-01-31
5R03DE014460-02 (2003): $78500
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Keith L Kirkwood
Regulated Insulin Delivery From Tissue Engineering Skin*
Keith L Kirkwood
University Of Michigan At Ann Arbor 3003 South State Street, Room 1040 Ann Arbor, Mi 481091274
Grant 5R01DK068673-02 from National Institute Of Diabetes And Digestive And Kidney Diseases IRG: ZDK1
Abstract: Type 1 diabetes is a disease characterized by lack of insulin due to autoimmune destruction of pancreatic beta-cells. Although insulin injections supply the amount of hormone necessary for survival, they often fail to provide tight regulation of blood glucose levels resulting in long-term complications such as neuropathy and cardiovascular disease. Here we propose to use gene therapy as an alternative therapeutic modality to supply constant baseline levels of insulin and regulate insulin secretion according to demand. Specifically, we propose to develop tissue engineered skin made of genetically modified keratinocytes as an in vivo delivery device to secrete insulin for the treatment of diabetes. We will construct retroviral vectors that regulate insulin secretion through exogenous administration of small organic molecules. Using optimal gene transfer protocols, we will transduce epidermal keratinocytes and examine secretion of active insulin from cells in culture and from stratified epidermal tissues. Transplantation of genetically modified skin equivalents onto athymic diabetic mice will be used to examine the effects of keratinocyte insulin secretion on blood glucose levels and determine the efficacy of insulin regulation in response to exogenous agents. Tissue engineered skin will be genetically modified to provide baseline amounts of insulin between meals and higher amounts before meals to mimic the insulin secretory response of beta-cells to nutrient stimulation. Our ultimate goal is to create a cell-based device to provide better control of blood glucose levels and overcome the devastating long-term effects of diabetes. Our team is composed of a tissue engineer (STA), a molecular biologist (KLK) and a diabetes expert (SGL). This combination of expertise is essential for the success of this multifaceted project, which involves molecular biology, tissue engineering, tissue transplantation and controlled drug delivery to engineer a cell-based device for regulated delivery of recombinant insulin.
Keywords: drug delivery system, gene therapy, insulin, insulin dependent diabetes mellitus, keratinocyte, nonhuman therapy evaluation, recombinant protein, skin transplantation, tissue engineering, blood glucose, athymic mouse, bioengineering /biomedical engineering, genetically modified animal
Project start date: 2004-08-05
Project end date: 2006-07-31
5R01DK068673-02 (2005): $76500
1R01DK068673-01 (2004): $180650
SOUTH CAROLINA COBRE FOR ORAL HEALTH RESEARCH
Keith L Kirkwood, Associate Dean For Research
Medical University Of South Carolina, 19 Hagood Ave., Suite 606, Charleston, Sc 29425
Grant 3P20RR017696-08S1 from National Center For Research Resources
Abstract: This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. To provide personnel and equipment for the Center for Oral Health Research to provide services to researchers engaged in oral cancer research
Project start date: 2009-08-26
Project end date: 2011-08-25
Budget start date: 26-AUG-2009
Budget end date: 25-AUG-2011
PFA/PA: RFA-RR-06-001
3P20RR017696-08S1 (2009): $651170
3P20RR017696-08S1_6840 (2009): $651170
3P20RR017696-08S2 (2009): $587298
3P20RR017696-08S2_6839 (2009): $587298
Keith L Kirkwood, Associate Dean For Research
Medical University Of South Carolina, 19 Hagood Ave., Suite 606, Charleston, Sc 29425
Abstract: This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This proposal seeks support for the development of the South Carolina COBRE for Oral Health at the Medical University of South Carolina (MUSC) College of Dental Medicine. The South Carolina COBRE for Oral Health will train a cadre of targeted investigators who will become established investigators and will provide academic leadership in the continued development of the College of Dental Medicine´s oral and craniofacial health research program. The program is highly leveraged with existing strengths at MUSC´s academic health science center including 1) a pool of outstanding faculty mentors who are collaborative and well-funded by NIH institutes and peer-review sponsors; 2) a pool of outstanding targeted investigators with strong, pre-existing relationships with members of the faculty mentor group; 3) critical relevant research space and core resources; 4) access to unique populations currently under investigation and served by MUSC´s community outreach programs; 5) access to clinical materials from these populations; and 6) an innovative collaborative relationship between MUSC and the NIDCR intramural research program on the NIH campus in Bethesda, MD that provides expanded research mentoring opportunities to targeted investigators in the South Carolina COBRE for Oral Health Program. Thus, the COBRE funding mechanism will allow the College of Dental Medicine to apply COBRE resources to most efficiently meet the goals of its research strategic plan to develop a nationally competitive and prominent oral health research program at MUSC which directly benefits the oral health care needs of the citizens of South Carolina. The South Carolina COBRE for Oral Health program will Mentor an initial cadre of five targeted investigators to develop successful, independent, NIH-funded research careers in oral health science. Enhance the oral health research infrastructure. COBRE funds will permit significantdevelopment of critical resources (cores) in the development of a research infrastructure supportive of multidisciplinary oral health research at MUSC´s academic health science center. Recruit two additional new faculty members whose experience and disciplinary expertise will complement those of the COBRE investigators in areas deemed critical by the Steering and the External Advisory Committees. Sustain the infrastructure of the South Carolina COBRE for Oral Health beyond the initial 5-year award
Keywords: Advisory Committees; Area; Award; COBRE; CRISP; Care, Health; Center of Biomedical Research Excellence; Centers of Research Excellence; Community Outreach; Complement; Complement Proteins; Computer Retrieval of Information on Scientific Projects Database; Dental; Development; Faculty; Funding; Funding Mechanisms; Goals; Grant; Health; Health Sciences; Healthcare; Infrastructure; Institutes; Institution; Intramural Program; Intramural Research Program; Investigation; Investigators; Leadership; Medical; Medicine; Mentors; NIDR; NIH; National Institute of Dental Research; National Institute of Dental and Craniofacial Research; National Institutes of Health; National Institutes of Health (U.S.); Oral Medicine; Oral health; Peer Review; Population; Programs (PT); Programs [Publication Type]; Recruitment Activity; Research; Research Infrastructure; Research Personnel; Research Resources; Researchers; Resources; Science of Medicine; Source; South Carolina; Stomatology; Strategic Planning; Task Forces; Training; United States National Institutes of Health; Universities; career; clinical material; college; craniofacial; craniofacies; dental health; experience; innovate; innovation; innovative; meetings; member; multidisciplinary; outreach program; programs; recruit
Project start date: 2009-06-01
Project end date: 2010-05-31
Budget start date: 1-JUN-2009
Budget end date: 31-MAY-2010
PFA/PA: RFA-RR-06-001
5P20RR017696-08_5949 (2009): $154032
MUSC DENTAL RESEARCH TRAINING GRANT
Keith L Kirkwood, Associate Dean For Research
Medical University Of South Carolina, 19 Hagood Ave., Suite 606, Charleston, Sc 29425
Grant 5T32DE017551-05 from National Institute Of Dental & Craniofacial Research
Abstract: This is a new application from the College of Dental Medicine at the Medical University of South Carolina (MUSC) that requests (1) continued NIDCR support for a summer research training program for dental students with a proven record of success; (2) support for a Dual Degree (DMD/PhD) Program established with institutional funds, which also has a proven record of success; and (3) support for a new Postdoctoral Research Program that will address a gap in a comprehensive roster of oral and craniofacial research training opportunities at MUSC. The full complement of research training options is enhanced by other NIH and institutionally supported programs at MUSC that offer research career development tracks for junior dental faculty (notably the NCRR P20 "COBRE" and NIDCR U24 awards) and an NIH Roadmap T32 award for predoctoral training in clinical research. Included in this proposal is a Progress Report summarizing relevant research training achievements to date as proof of principle and documentation. The overarching objective of the proposed Dental Research Training Program (DRTP) is to train an interdisciplinary cadre of talented investigators for careers in dental, oral and craniofacial science who will become the next generation of dental school faculty and provide academic leadership in the nation´s dental, oral and craniofacial research programs. Mentoring and training activities are thematically grouped in five strategic areas of research focus (i) infection and immunity, (ii) oral and systemic health, (iii) oral cancer, (iv) regenerative medicine/bioengineering, and (v) proteomics. Integrative training components, including an Oral Health Journal Club, Seminar Series, and Student Research Day, provide a framework for all trainees to interact, collaborate, and work together during their training experience. The proposed program benefits from the excellent administrative support and institutional commitment already in place and operating effectively in the Center for Oral Health Research. An integrated multi-year evaluation plan will be used to measure the extent to which the program goals and objectives are met
Keywords: Dental Research; Grant; Research Training
Project start date: 2006-07-01
Project end date: 2011-06-30
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2011
PFA/PA: PAR-05-101
5T32DE017551-05 (2010): $451298
5T32DE017551-04 (2009): $467659
5T32DE017551-02 (2007): $343084
Sponsored Links Excellgen http://Excellgen.com
SOUTH CAROLINA COBRE FOR ORAL HEALTH RESEARCH
Keith L Kirkwood, Associate Dean For Research
Medical University Of South Carolina, 19 Hagood Ave., Suite 606, Charleston, Sc 29425
Grant 5P20RR017696-09 from National Center For Research Resources
Abstract: This application from the College of Dental Medicine (COM) at the Medical University of South Carolina (MUSC) requests renewal of the COBRE for Oral Health. The initial COBRE award in 2002 had a catalytic impact on the development of research capacity in MUSC´s College of Dental Medicine and dramatically enlightened awareness of and interest in oral and craniofacial research campus-wide. The overall objective of this proposal is to firmly establish the MUSC Center for Oral Health Research (COHR) as a sustainable, independent multidisciplinary research center. Continued funding through the NCRR COBRE Program will ensure that the momentum gained during the first cycle will be unabated and the trajectory of research competitiveness will continue to climb in terms of both individual investigator awards and multi-investigator mechanisms such as program projects, center grants and additional training grants. In this renewal proposal we are requesting support for five junior investigators (four currently at MUSC and one to be recruited), who will conduct research projects of their own design with substantial one on- one and team mentoring, as well as two critical scientific cores and an administrative core that includes proven components for management and oversight, faculty development and program planning. The specific aims for the COBRE renewal are AIM 1 Expand the critical mass of funded investigators in oral and craniofacial research by a. mentoring the research career development of 5 to 6 additional outstanding new targeted investigators with interests and talents relevant to our theme, and b. enhancing the long-term development and continuing success of graduates as independent investigators. AIM 2 Enhance scientific core resources that will serve as platforms to increase the capacity of our targeted faculty to compete successfully for NIH funding. AIM 3 Fully implement the MUSC Center for Oral Health Research as the vehicle for successful transition to long-term competitive, renewable support from a diverse range of federal and non-federal sources
Project start date: 2002-09-30
Project end date: 2012-05-31
Budget start date: 1-JUN-2010
Budget end date: 31-MAY-2011
PFA/PA: RFA-RR-06-001
5P20RR017696-09 (2010): $2147085
5P20RR017696-08 (2009): $2147085
NEGATIVE REGULATION OF INNATE IMMUNE RESPONSES THROUGH MAPK PHOSPHATASES
Keith L Kirkwood, Associate Dean For Research
Medical University Of South Carolina, 19 Hagood Ave., Suite 606, Charleston, Sc 29425
Grant 5R01DE018290-05 from National Institute Of Dental & Craniofacial Research
Abstract: Periodontal disease initiation and progression occurs as a consequence of the host immune inflammatory response to oral pathogens. The production of inflammatory cytokines is a highly regulated process involving transcriptional and posttranscriptional mechanisms. One of the major signaling pathways activated by periopathogenic LPS in p38 MARK. Following p38 phosphorylation, inactivation of p38 MAP kinases is achieved mainly by a family of dual-specific MAP kinase phosphatases (MKP). MKP-1 is capable of negatively regulating both transcriptional and post transcriptional p38 MAP kinase activity. MKP-1 contributes towards LPS tolerance and over-expression of MKP-1 has shown to accelerate p38 inactivation resulting in diminished proinflammatory cytokine production. We have recently shown that LPS-induced IL-6 mRNA stability expression requires p38 signaling. Preliminary data for this proposal indicates that in MKP- 1 transfected cells, LPS-induced IL-6 expression is significantly attenuated. In addition, we have provided significant data showing the p38 is a major signaling pathway contributing to LPS-induced periodontal bone destruction. Based upon these data, we hypothesize that the endogenous negative regulator mechanism of p38 signaling, MKP-1, is a key component of responsible for attenuation of LPS-induced inflammatory cytokine expression in macrophages. In this proposal, the ability of TIP over-expression to decrease inflammation will be determined in vitro using gene targeted strategies in macrophages, and in vivo using experimental periodontitis models. The specific aims are 1) To determine the role of over-expressed MKP-1 on IL-6 and TNFa mRNA expression in vitro. 2) To determine the contribution of MKP-1 in ontogeny of inflammatory cytokine production and LPS-induced osteoclastogenesis in primary bone marrow macrophages and 3) To determine the impact of MKP-1 in inflammatory bone destruction in vivo using MKP mice. These studies will establish the role of LPS-induced cytokine expression and negative regulation in inflammatory bone loss through selective attenuation of p38 MAPK-induced signaling in periodontal bone destruction
Keywords: 21+ years old; 3` Untranslated Regions; 3`UTR; A. actinomycetemcomitans; ATP-protein phosphotransferase; Abbreviations; Acid Phosphatase; Actinobacillus actinomycetemcomitans; Acute; Address; Adenosine; Adult; Alveolar Bone Loss; Alveolar Resorption; Assay; Attenuated; B cell differentiation factor; B cell stimulating factor 2; B-Cell Differentiation Factor-2; B-Cell Stimulatory Factor-2; BCDF; BRF Gene; BRF1; BRF1 gene; BRF2; BSF-2; BSF2; BSF2 (B cell stimulating factor 2); Bacterium actinomycetem comitans; Bacterium comitans; Binding Proteins; Bioassay; Biologic Assays; Biological Assay; Body Tissues; Bone; Bone Marrow; Bone and Bones; Bones and Bone Tissue; Butyrate Response Factor 2; CAT Scan, X-Ray; CAT scan; COX-2; COX-2 protein; COX2; COX2 enzyme; CSBP1; CSBP2; CSPB1; CT X Ray; CT scan; Cachectin; Cachectin-Tumor Necrosis Factor; Cell Communication and Signaling; Cell Signaling; Cell/Tissue, Immunohistochemistry; Cells; Chronic; Chronic Periodontitis; Common Rat Strains; Computed Tomography; Computerized Axial Tomography (Computerized Tomography); Computerized Tomography, X-Ray; Cyclo-Oxygenase-2; Cyclooxygenase; Cytokines and Inflammatory Response; Data; Development; Differentiation Factor, B-Cell; Dinoprostone; Disease; Disease Progression; Disorder; EC 2.7; EC 2.7.2-; EGF-Response Factor 2; EGF-response factor 2 protein, human; EMI scan; ERF-2 Protein; ERF-2 protein, human; ERF2; ERK MAP Kinases; EXIP; Elements; Experimental Models; Experimental Models, Other; Extracellular Signal Regulated Kinases; Extracellular Signal-Regulated Kinase Gene; Extracellular Signal-Regulated Kinases; Extracellular Signal-Regulated MAP Kinases; Family; G0-G1 switch regulatory protein 24; GFP; GM-CSF; GMCSF; GOS24 protein; Gene Targeting; General Transcription Factor 3B, 90-kD Subunit; Generalized Growth; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Green Fluorescent Proteins; Growth; HPGF; Hepatocyte-Stimulating Factor; Histamine-Producing Cell-Stimulating Factor; Human, Adult; Hybridoma Growth Factor; IFN-beta 2; IFNB2; IHC; IL-6; IL6 Protein; INFLM; Immune; Immune response; Immunoglobulin Enhancer-Binding Protein; Immunohistochemistry; Immunohistochemistry Staining Method; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Inflammatory Response Pathway; Interleukin 6 (Interferon, Beta 2); Interleukin-6; Interleukins; Intracellular Communication and Signaling; Investigators; JN Kinase; JNK; JNK Mitogen-Activated Protein Kinases; JNK1; JNK1 Kinase; JNK1 protein; JNK1A2; JNK21B1/2; Kinases; LPS; Ligand Binding Protein; Lipopolysaccharides; MAP Kinase 8; MAP Kinase 8 Gene; MAP Kinase Gene; MAP kinase; MAPK; MAPK phosphatase; MAPK14; MAPK14 gene; MAPK8; MAPK8 Mitogen-Activated Protein Kinase; MAPK8 gene; MGI-2; MMP-13; MMP-13 gene product; MMP13 gene product; MMPs; Mammals, Mice; Mammals, Rats; Matrix Metalloproteinases; Mice; Mitogen-Activated Protein Kinase 8; Mitogen-Activated Protein Kinase Gene; Mitogen-Activated Protein Kinases; Modeling; Models, Experimental; Molecular; Molgramostin; Murine; Mus; Mxi2; Myeloid Differentiation-Inducing Protein; NF-kB; NF-kappa B; NF-kappaB; NFKB; Nature; NuP475 protein; Nuclear Factor kappa B; Nuclear Transcription Factor NF-kB; ODF; OPGL; Oral; Organism; Orthophosphoric-monoester phosphohydrolase (acid optimum); Osteoclasts; PGE2; PGE2 alpha; PGE2alpha; PGG/HS; PGH Synthase 2; PGHS-2; PGHS2; PHS II; PHS-2; PRKM14; PRKM15; PRKM8; PTGS2; PTGS2 gene; Parodontosis; Pathway interactions; Periodontal Bone Loss; Periodontal Diseases; Periodontal Resorption; Periodontitis; Phosphatases; Phosphohydrolases; Phosphomonoesterases; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Plasmacytoma Growth Factor; Play; Process; Production; Programs (PT); Programs [Publication Type]; Prosta-5, 13-dien-1-oic acid, 11, 15-dihydroxy-9-oxo-, (5Z, 11alpha, 13E, 15S)-; Prostaglandin E2; Prostaglandin E2 alpha; Prostaglandin E2alpha; Prostaglandin G/H Synthase 2; Prostaglandin G/H Synthase and Cyclooxygenase; Prostaglandin H2 Synthase 2; Prostaglandin-Endoperoxide Synthase 2; Protein Kinase; Protein Phosphorylation; RANKL; Rat; Rattus; Receptor Protein; Regulation; Research Personnel; Researchers; Resistance; Reticuloendothelial System, Bone Marrow; Role; SAP Kinase-1; SAPK/JNK; SAPK1; SAPK1 Mitogen-Activated Protein Kinase; SAPK1/JNK; SAPK2A; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Site; Stability, mRNA; Stress; Stress-Activated Protein Kinase JNK1; Stress-Activated Protein Kinase gamma; System; System, LOINC Axis 4; TAF3B2 Gene; TAF3C, Formerly; TC-GM-CSF; TFIIIB90 Gene; TIS11 protein; TIS11D; TIS11D Protein; TIS11b protein; TLR protein; TNF (unspecified); TNF Receptor Ligands; TNF-alpha; TNFSF11; TNFSF11 gene; TTP protein; Targetings, Gene; Threonine/Tyrosine Protein Kinase; Tissue Growth; Tissues; Toll-like receptors; Tomodensitometry; Tomography, Xray Computed; Transcription Factor NF-kB; Transphosphorylases; Tumor Necrosis Factor; Tumor Necrosis Factor Family Protein; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Tumor-Cell Human GM Colony-Stimulating Factor; Urd; Uridine; Wild Type Mouse; X-Ray Computed Tomography; ZFP36 protein; ZFP36L2; ZFP36L2 gene; ZFP36L2 protein, human; Zinc Finger Protein 36 C3H Type-Like 2; adult human (21+); attenuation; base; biological signal transduction; bone; bone loss; butyrate response factor 1; butyrate response factor 2 protein, human; c-jun Amino-Terminal Kinase; c-jun Kinase-1; c-jun N-Terminal Kinase; c-jun N-Terminal Kinase 1; cMG1 protein; catscan; collagenase 3; computed axial tomography; computerized axial tomography; computerized tomography; cyclo-oxygenase II; cyclooxygenase 2; cytokine; disease/disorder; extracellular signal related kinase; fetal; glycogen synthase a kinase; granulocyte macrophage colony stimulating factor; hCOX-2; hRANKL2; host response; human ZFP36L2 protein; hydroxyalkyl protein kinase; immunoresponse; in vivo; interferon beta 2; jun-NH2-Terminal Kinase; kappa B Enhancer Binding Protein; living system; mRNA Expression; mRNA Stability; macrophage; matrix metalloproteinase-13; mitogen-activated protein kinase p38; nuclear factor kappa beta; ontogeny; oral pathogen; osteoclastogenesis; p38; p38 MAP Kinase; p38 MAPK; p38 MAPK Gene; p38 Protein Kinase; p38 SAPK; p38Alpha; pathogen; pathway; periodontal disorder; periodontium disease; periodontium disorder; phosphorylase b kinase kinase; programs; prostaglandin H synthase-2; prototype; receptor; resistant; response; sOdf; social role; stress-activated protein kinase 1; tristetraprolin; tumor necrosis factor (unspecified); unspecified interleukin; zinc finger protein 36, C3H type-like 2 protein, human
Project start date: 2007-02-07
Project end date: 2012-01-31
Budget start date: 1-FEB-2010
Budget end date: 31-JAN-2011
PFA/PA: RFA-DE-07-004
5R01DE018290-05 (2010): $332163
3R01DE018290-04S2 (2009): $99675
1R01DE018290-01 (2007): $347208
ALTERING CYTOKINE MRNA STABILITY IN PERIODONTAL BONE LOSS
Keith L Kirkwood, Associate Dean For Research
Medical University Of South Carolina, 19 Hagood Ave., Suite 606, Charleston, Sc 29425
Grant 5R21DE017966-03 from National Institute Of Dental & Craniofacial Research
Abstract: Periodontal disease initiation and progression occurs as a consequence of the host immune inflammatory response to oral pathogens. The production of inflammatory cytokines is a highly regulated process involving transcriptional and posttranscriptional mechanisms. At the posttranscriptional level, the presence of adenosine/uridine-rich elements (AREs) in the 3´ untranslated region of many cytokine genes, including IL-6, TNF-a, and COX-2, targets the mRNA for rapid degradation. As a consequence, cytokine production is repressed. Tristetraprolin (TTP) is a zinc finger protein that binds to the ARE of cytokine mRNAs and enhances degradation of the mRNA. TTP is phosphorylated by the p38-MK2 pathway and may serve as a general mechanism of cytokine mRNA regulation. We have recently shown that LPS-induced IL-6 mRNA stability expression requires p38 signaling. Preliminary data for this proposal indicates that TTP transfected cells inhibit LPS-induced IL-6 expression. Based upon these data, we hypothesize that the mRNA decay enhancing properties of TTP may be exploited for potential anti-inflammatory purposes. In this proposal, the ability of TTP over-expression to decrease inflammation will be determined in vitro using gene targeted strategies in macrophages, and in vivo using experimental periodontitis models. The specific aims are 1) to clarify the role of TTP on IL-6, TNF, and COX-2 mRNA expression and stability in vitro and 2) determine the impact of TTP in inflammatory and periodontal pathogen- initiated bone destruction in vivo using TTP-/- mice. These studies will establish the role of LPS-induced cytokine mRNA stability in inflammatory bone loss through selective mRNA decay targeting with TTP. Upon the accomplishment of these aims, subsequent studies will address the potential of TTP as a therapeutic strategy to control periodontal bone loss in small animal models. Periodontal disease initiation and progression occurs as a consequence of the host immune inflammatory response to oral pathogens. These studies will establish the role of host immune proteins termed cytokines and the regulation of cytokines at the level of mRNA stability in inflammatory bone loss through selective mRNA decay targeting with a protein that regulates cytokine mRNA stability. Progress in understanding the role of posttranscriptional cytokine regulation in periodontal inflammation and bone loss may yield new possibilities for treatment of periodontal diseases and other chronic inflammatory diseases
Keywords: 3` Untranslated Regions; 3`UTR; Address; Adenosine; Alveolar Bone Loss; Alveolar Resorption; Animal Model; Animal Models and Related Studies; Anti-Inflammatories; Anti-Inflammatory Agents; Anti-inflammatory; Antiinflammatories; Antiinflammatory Agents; B cell differentiation factor; B cell stimulating factor 2; B-Cell Differentiation Factor-2; B-Cell Stimulatory Factor-2; BCDF; BSF-2; BSF2; BSF2 (B cell stimulating factor 2); Bone; Bone and Bones; Bones and Bone Tissue; CAT Scan, X-Ray; CAT scan; COX-2; COX2; CSBP1; CSBP2; CSPB1; CT X Ray; CT scan; Cell Communication and Signaling; Cell Signaling; Cells; Chronic; Chronic Periodontitis; Complex; Computed Tomography; Computerized Axial Tomography (Computerized Tomography); Computerized Tomography, X-Ray; Cytokine Gene; DIF; Data; Decay, mRNA; Degradation, mRNA; Differentiation Factor, B-Cell; Dinoprostone; Disease; Disorder; EC 2.7; EMI scan; EXIP; Elements; G0-G1 switch regulatory protein 24; GOS24 protein; Gene Targeting; HPGF; Hepatocyte-Stimulating Factor; Hybridoma Growth Factor; IFN-beta 2; IFNB2; IL-1; IL-6; IL1; IL6 Protein; INFLM; Immune; In Vitro; In element; Indium; Inflammation; Inflammatory; Inflammatory Response; Interleukin 6 (Interferon, Beta 2); Interleukin I; Interleukin-1; Interleukin-6; Intracellular Communication and Signaling; Kinases; Lymphocyte-Stimulating Hormone; MAPK14; MAPK14 gene; MGI-2; Macrophage Cell Factor; Mammals, Mice; Membrane; Messenger RNA; Mice; Modeling; Murine; Mus; Mxi2; Myeloid Differentiation-Inducing Protein; NuP475 protein; ODF; OPGL; Oral; PGE2; PGE2 alpha; PGE2alpha; PGG/HS; PGHS-2; PHS-2; PRKM14; PRKM15; PTGS2; PTGS2 gene; Parodontosis; Pathway interactions; Periodontal Bone Loss; Periodontal Diseases; Periodontal Resorption; Periodontitis; Phosphotransferases; Plasmacytoma Growth Factor; Plasmids; Process; Production; Property; Property, LOINC Axis 2; Prosta-5, 13-dien-1-oic acid, 11, 15-dihydroxy-9-oxo-, (5Z, 11alpha, 13E, 15S)-; Prostaglandin E2; Prostaglandin E2 alpha; Prostaglandin E2alpha; Protein Binding; Proteins; Purpose; RANKL; RNA, Messenger; Regulation; Role; SAPK2A; Signal Transduction; Signal Transduction Systems; Signaling; Stability, mRNA; Stress; T Helper Factor; TIS11 protein; TNF; TNF A; TNF gene; TNFSF11; TNFSF11 gene; TNFSF2; TTP protein; Targetings, Gene; Therapeutic; Tomodensitometry; Tomography, Xray Computed; Transphosphorylases; Tumor Necrosis Factor Gene; Urd; Uridine; X-Ray Computed Tomography; ZFP36 protein; Zinc Finger Domain; Zinc Finger Motifs; Zinc Fingers; base; biological signal transduction; bone; bone loss; catscan; computed axial tomography; computerized axial tomography; computerized tomography; cytokine; disease/disorder; gene product; hCOX-2; hRANKL2; in vivo; interferon beta 2; lymphocyte activating factor; mRNA; mRNA Decay; mRNA Expression; mRNA Stability; mRNA Transcript Degradation; macrophage; membrane structure; model organism; oral pathogen; p38; p38 MAPK Gene; p38Alpha; pathogen; pathogenic bacteria; pathway; periodontal disorder; periodontium disease; periodontium disorder; sOdf; social role; tristetraprolin
Project start date: 2007-07-01
Project end date: 2010-06-30
Budget start date: 1-JUL-2008
Budget end date: 30-JUN-2010
PFA/PA: PA-06-181
5R21DE017966-03 (2008): $0
1R21DE017966-01A1 (2007): $182535
SILENCING POST TRANSCRIPTIONAL SIGNALING MECHANISMS IN PERIODONTAL INFLAMMATION
Keith L Kirkwood, Associate Dean For Research
Medical University Of South Carolina, 19 Hagood Ave., Suite 606, Charleston, Sc 29425
Grant 5R21DE019272-02 from National Institute Of Dental & Craniofacial Research
Abstract: Periodontal disease initiation and progression occurs as a consequence of the host immune inflammatory response to oral pathogens. The production of inflammatory cytokines is a highly regulated process involving transcriptional and post transcriptional mechanisms. At the post transcriptional level, the presence of adenosine/uridine-rich elements (AREs) in the 3´ untranslated region of many cytokine genes, including IL-6, TNF1, and COX-2, targets the mRNA for rapid degradation. As a consequence, cytokine production is repressed. Tristetraprolin (TTP) is a zinc finger protein that binds to the ARE of cytokine mRNAs and enhances degradation of the mRNA. TTP is phosphorylated by the p38-MK2 pathway and may serve as a general mechanism of cytokine mRNA regulation. We have recently shown that LPS-induced bone loss in vivo requires p38 signaling in experimental periodontitis. Preliminary data for this proposal indicates that when MK2 is silenced through siRNA significant reduction in LPS-induced cytokine expression is observed. Based upon these data, we hypothesize that selective knockdown of MK2 may offer significant therapeutic benefit in inflammatory periodontal diseases. In this proposal, MK2 siRNA will be evaluated to decrease inflammation in vitro using gene-targeted strategies in macrophages, and in vivo using experimental periodontitis models. The Specific Aims are 1) validation of MK2 silencing in cytokine production and posttranscriptional regulation; and 2) in vivo evaluation of MK2 knockdown in an experimental periodontitis model. These studies will determine if siRNA to MK2 can be delivered intraorally as a novel therapeutic that selectively targets cytokine mRNA stability in inflammatory bone loss. Upon accomplishment of these aims, subsequent studies will address optimization of MK2 siRNA, a therapeutic strategy to control periodontal bone loss in infectious animal models of periodontitis. Periodontal disease progression occurs as a consequence of the host immune inflammatory response to oral pathogens. These studies will establish the role of host immune proteins termed cytokines and the regulation of cytokines by selectively silencing a key signaling molecule needed to mediate inflammatory cytokine mRNA stability. Using established small animal models of experimental periodontitis, we will evaluate the ability of silencing this signaling protein for therapeutic benefit. Progress in understanding the role of posttranscriptional cytokine regulation in periodontal inflammation and bone loss may yield new possibilities for treatment of periodontal diseases and other chronic inflammatory diseases
Keywords: 3` Untranslated Regions; 3`UTR; A. actinomycetemcomitans; ATGN; ATP-protein phosphotransferase; Abbreviations; Acid Phosphatase; Actinobacillus; Actinobacillus actinomycetemcomitans; Address; Adenosine; Alveolar Bone Loss; Alveolar Resorption; Animal Model; Animal Models and Related Studies; Anti-Inflammatories; Anti-Inflammatory Agents; Anti-inflammatory; Antigens; Antiinflammatories; Antiinflammatory Agents; Assay; B cell differentiation factor; B cell stimulating factor 2; B-Cell Differentiation Factor-2; B-Cell Stimulatory Factor-2; BCDF; BRF Gene; BRF1; BRF1 gene; BRF2; BSF-2; BSF2; BSF2 (B cell stimulating factor 2); Bacterium actinomycetem comitans; Bacterium comitans; Binding Proteins; Bio-Informatics; Bioassay; Bioinformatics; Biologic Assays; Biological Assay; Bone Marrow; Buccal Mucosa; Butyrate Response Factor 2; CAT Scan, X-Ray; CAT scan; COX-2; COX-2 protein; COX2; COX2 enzyme; CSBP1; CSBP2; CSPB1; CT X Ray; CT scan; Cachectin; Cachectin-Tumor Necrosis Factor; Cell Communication and Signaling; Cell Signaling; Cell/Tissue, Immunohistochemistry; Cells; Chronic; Chronic Periodontitis; Common Rat Strains; Computed Tomography; Computerized Axial Tomography (Computerized Tomography); Computerized Tomography, X-Ray; Conserved Sequence; Cyclo-Oxygenase-2; Cyclooxygenase; Cytokine Gene; DIF; Data; Decay, mRNA; Degradation, mRNA; Differentiation Factor, B-Cell; Dinoprostone; Disease; Disease Progression; Disorder; EC 2.7; EC 2.7.2-; EGF-Response Factor 2; EGF-response factor 2 protein, human; ELISA; EMI scan; ERF-2 Protein; ERF-2 protein, human; ERF2; EXIP; Elements; Enzyme-Linked Immunosorbent Assay; Evaluation; Experimental Animal Model; Experimental Models; Experimental Models, Other; Extracellular Signal-Regulated Kinase Gene; Extracellular Signal-Regulated Kinases; G0-G1 switch regulatory protein 24; GFP; GM-CSF; GMCSF; GOS24 protein; Gene Products, RNA; Gene Targeting; General Transcription Factor 3B, 90-kD Subunit; Granulocyte-Macrophage Colony-Stimulating Factor; Green Fluorescent Proteins; HPGF; Hepatocyte-Stimulating Factor; Histamine-Producing Cell-Stimulating Factor; Human; Human, General; Hybridoma Growth Factor; IFN; IFN-beta 2; IFNB2; IHC; IL-1; IL-6; IL1; IL6 Protein; INFLM; Immune; Immune response; Immunoglobulin Enhancer-Binding Protein; Immunohistochemistry; Immunohistochemistry Staining Method; In Vitro; In element; Indium; Inflammation; Inflammatory; Inflammatory Response; Interferons; Interleukin 6 (Interferon, Beta 2); Interleukin I; Interleukin-1; Interleukin-6; Intracellular Communication and Signaling; Kinases; LPS; Label; Ligand Binding Protein; Lipopolysaccharides; Lymphocyte-Stimulating Hormone; MAP Kinase Gene; MAP kinase; MAPK; MAPK14; MAPK14 gene; MGI-2; MMP-13; MMP-13 gene product; MMP13 gene product; MMPs; Macrophage Cell Factor; Mammals, Mice; Mammals, Rats; Man (Taxonomy); Man, Modern; Matrix Metalloproteinases; Measures; Mediating; Membrane; Messenger RNA; Mice; Microinjections; Mitogen-Activated Protein Kinase Gene; Mitogen-Activated Protein Kinases; Modeling; Models, Experimental; Molgramostin; Mouth Mucosa; Murine; Mus; Mxi2; Myeloid Differentiation-Inducing Protein; NF-kB; NF-kappa B; NF-kappaB; NFKB; NuP475 protein; Nuclear Factor kappa B; Nuclear Transcription Factor NF-kB; ODF; OPGL; Oral; Oral Mucosa; Oral mucous membrane structure; Orthophosphoric-monoester phosphohydrolase (acid optimum); PGE2; PGE2 alpha; PGE2alpha; PGG/HS; PGH Synthase 2; PGHS-2; PGHS2; PHS II; PHS-2; PRKM14; PRKM15; PTGS2; PTGS2 gene; Parodontosis; Pathway interactions; Periodontal Bone Loss; Periodontal Diseases; Periodontal Resorption; Periodontitis; Phosphotransferases; Plasmacytoma Growth Factor; Process; Production; Prosta-5, 13-dien-1-oic acid, 11, 15-dihydroxy-9-oxo-, (5Z, 11alpha, 13E, 15S)-; Prostaglandin E2; Prostaglandin E2 alpha; Prostaglandin E2alpha; Prostaglandin G/H Synthase 2; Prostaglandin G/H Synthase and Cyclooxygenase; Prostaglandin H2 Synthase 2; Prostaglandin-Endoperoxide Synthase 2; Protein Binding; Protein Kinase; Proteins; RANKL; RNA; RNA Splicing; RNA Stability; RNA, Messenger; RNA, Non-Polyadenylated; RNA, Small Interfering; Rat; Rattus; Receptor Protein; Regulation; Regulatory Protein; Reporter; Resistance; Reticuloendothelial System, Bone Marrow; Ribonucleic Acid; Role; SAPK2A; Signal Transduction; Signal Transduction Systems; Signaling; Signaling Molecule; Signaling Protein; Small Interfering RNA; Specificity; Splicing; Stability, mRNA; Stress; System; System, LOINC Axis 4; T Helper Factor; T-Cells; T-Lymphocyte; TAF3B2 Gene; TAF3C, Formerly; TC-GM-CSF; TFIIIB90 Gene; TIS11 protein; TIS11D; TIS11D Protein; TIS11b protein; TLR protein; TNF; TNF (unspecified); TNF A; TNF Receptor Ligands; TNF gene; TNF-alpha; TNFSF11; TNFSF11 gene; TNFSF2; TTP protein; Targetings, Gene; Therapeutic; Thymus-Dependent Lymphocytes; Toll-like receptors; Tomodensitometry; Tomography, Xray Computed; Transcription Factor NF-kB; Translations; Transphosphorylases; Tumor Necrosis Factor; Tumor Necrosis Factor Family Protein; Tumor Necrosis Factor Gene; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Tumor-Cell Human GM Colony-Stimulating Factor; Urd; Uridine; Validation; X-Ray Computed Tomography; ZFP36 protein; ZFP36L2; ZFP36L2 gene; ZFP36L2 protein, human; Zinc Finger Domain; Zinc Finger Motifs; Zinc Finger Protein 36 C3H Type-Like 2; Zinc Fingers; base; biological signal transduction; bone loss; butyrate response factor 1; butyrate response factor 2 protein, human; cMG1 protein; carboxyfluorescein; catscan; collagenase 3; computed axial tomography; computerized axial tomography; computerized tomography; cyclo-oxygenase II; cyclooxygenase 2; cytokine; design; designing; disease/disorder; gene product; genetic regulatory protein; glycogen synthase a kinase; granulocyte macrophage colony stimulating factor; hCOX-2; hRANKL2; host response; human ZFP36L2 protein; hydroxyalkyl protein kinase; immunogen; immunoresponse; in vitro Assay; in vivo; interferon beta 2; kappa B Enhancer Binding Protein; lymphocyte activating factor; mRNA; mRNA Decay; mRNA Stability; mRNA Transcript Degradation; macrophage; matrix metalloproteinase-13; membrane structure; model organism; new therapeutics; next generation therapeutics; novel; novel therapeutics; nuclear factor kappa beta; oral mucosae; oral mucosal; oral pathogen; p38; p38 MAPK Gene; p38Alpha; pathogenic bacteria; pathway; periodontal disorder; periodontium disease; periodontium disorder; phosphorylase b kinase kinase; prostaglandin H synthase-2; protein complex; receptor; regulatory gene product; resistant; sOdf; siRNA; social role; thymus derived lymphocyte; tristetraprolin; tumor necrosis factor (unspecified); zinc finger protein 36, C3H type-like 2 protein, human
Project start date: 2008-09-01
Project end date: 2010-08-31
Budget start date: 1-SEP-2009
Budget end date: 31-AUG-2010
PFA/PA: RFA-DE-08-005
5R21DE019272-02 (2009): $221250
Sponsored Links Excellgen http://Excellgen.com
INSP3R GENE REGULATION IN OSTEOBLASTS BY 17-B ESTRADIOL
Keith L Kirkwood
Pharmacology And Toxicologystate University Of New York At Buffalo
sponsored Projects Services
buffalo, Ny 14260
Grant 5F32DE005697-02 from National Institute Of Dental & Craniofacial Research IRG: ORTH
5F32DE005697-02 (1997): $36300
1F32DE005697-01 (1996): $35300