The long-term objective of this proposal is to understand the mechanisms responsible for the termination of proinflammatory cytokine biosynthesis in macrophages. Proinflammatory cytokines, TNF-alpha and IL-1 in particular, play an important role in the pathogenesis of a variety of human diseases, including rheumatoid arthritis, Crohn's disease, and septic shock. Biosyntheses of both TNF-a and IL-1 in LPS-stimulated macrophages are regulated by complex signal transduction pathways involving MAP kinases and NF-kappaB. Preliminary studies in our laboratory with RAW264.7 cells provide strong evidence to support the hypothesis that MKP-1 plays a critical role in the feedback control of p38 and JNK MAP kinases and is responsible for the termination of pro-inflammatory cytokine production in LPS-stimulated macrophages. Moreover, MKP-1 is potently induced by glucocorticoids. The First Specific Aim of the present proposal is to test the hypothesis that MKP-1 plays critical roles in restraining the LPS-induced biosynthesis of proinflarnmatory cytokines in both peritoneal and alveolar macrophages after LPS stimulation. The Second Specific Aim is to test the hypothesis that MKP-1 also acts as a critical negative regulator in the restraint of macrophage responses to Gram-positive bacteria. The Third Specific Aim is to test the hypothesis that the responses to LPS stimulation of primary peritoneal macrophages from Mkp-1-/- mice differ from the responses of macrophages isolated from Mkp-l+/+ mice. The cryopreserved embryos derived from the Mkp-1 knockout mice are obtained from Bristol-Myers Squibb Pharmaceutical Research Institute, through a materials transfer agreement. Regeneration of these embryos into mice has been accomplished in The Jackson Laboratory. We will isolate peritoneal macrophages from the Mkp-l+/+ and Mkp-1-/- mice and use these macrophages to determine the role of MKP- 1 in the responses to LPS, with respect to MAP kinase inactivation and control of inflammatory cytokine production. We will also examine whether the lack of Mkp-1 gene will compromise the suppressant effects of glucocorticoids on TNF-alpha production induced by LPS. These studies are designed to answer pivotal questions regarding the regulatory mechanisms responsible for terminating proinflammatory cytokine production in macrophages during bacterial infections, and to reveal novel targets for developing new antiinflammatory/ anti-rheumatic drugs.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI057798-02
Application #
6889491
Study Section
Special Emphasis Panel (ZRG1-SSS-F (01))
Program Officer
Dong, Gang
Project Start
2004-05-01
Project End
2009-02-28
Budget Start
2005-03-01
Budget End
2006-02-28
Support Year
2
Fiscal Year
2005
Total Cost
$292,000
Indirect Cost
Name
Nationwide Children's Hospital
Department
Type
DUNS #
147212963
City
Columbus
State
OH
Country
United States
Zip Code
43205
Crowell, Sara; Wancket, Lyn M; Shakibi, Yasmine et al. (2014) Post-translational regulation of mitogen-activated protein kinase phosphatase (MKP)-1 and MKP-2 in macrophages following lipopolysaccharide stimulation: the role of the C termini of the phosphatases in determining their stability. J Biol Chem 289:28753-64
Yan, Jing; Ralston, Melissa M; Meng, Xiaomei et al. (2013) Glutathione reductase is essential for host defense against bacterial infection. Free Radic Biol Med 61:320-32
Wancket, Lyn M; Meng, Xiaomei; Rogers, Lynette K et al. (2012) Mitogen-activated protein kinase phosphatase (Mkp)-1 protects mice against acetaminophen-induced hepatic injury. Toxicol Pathol 40:1095-105
Vandevyver, Sofie; Dejager, Lien; Van Bogaert, Tom et al. (2012) Glucocorticoid receptor dimerization induces MKP1 to protect against TNF-induced inflammation. J Clin Invest 122:2130-40
Frazier, W Joshua; Xue, Jianjing; Luce, Wendy A et al. (2012) MAPK signaling drives inflammation in LPS-stimulated cardiomyocytes: the route of crosstalk to G-protein-coupled receptors. PLoS One 7:e50071
Matta, Ranyia; Barnard, John A; Wancket, Lyn M et al. (2012) Knockout of Mkp-1 exacerbates colitis in Il-10-deficient mice. Am J Physiol Gastrointest Liver Physiol 302:G1322-35
Zhang, Ting; Lu, Xiangru; Arnold, Paul et al. (2012) Mitogen-activated protein kinase phosphatase-1 inhibits myocardial TNF-? expression and improves cardiac function during endotoxemia. Cardiovasc Res 93:471-9
Wancket, Lyn M; Frazier, W Joshua; Liu, Yusen (2012) Mitogen-activated protein kinase phosphatase (MKP)-1 in immunology, physiology, and disease. Life Sci 90:237-48
Yan, Jing; Meng, Xiaomei; Wancket, Lyn M et al. (2012) Glutathione reductase facilitates host defense by sustaining phagocytic oxidative burst and promoting the development of neutrophil extracellular traps. J Immunol 188:2316-27
Zhang, Yong; Leung, Donald Y M; Richers, Brittany N et al. (2012) Vitamin D inhibits monocyte/macrophage proinflammatory cytokine production by targeting MAPK phosphatase-1. J Immunol 188:2127-35

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