This is a competitive renewal of a grant, which has for several years examined the mechanisms of regulation of prostaglandin endoperoxide synthase (PGHS). Glomerular mesangial cells (GMC) play a key role in the genesis of several glomerular diseases such as glomerulosclerosis and proliferative glomerulonephritis, which are characterized by GMC injury and proliferation. Proinflammatory agents can induce cellular synthesis of eicosanoids, namely oxygenated products of arachidonic acid, that can regulate GMC matrix synthesis and proliferation. This application examines the intracellular signaling mechanisms regulating the expression and actions of PGHS, which catalyses prostaglandin production from arachidonic acid. PGHS is of particular interest, being the principal target of NSAIDS. A combination of in vitro and in vivo studies will be used to test the hypothesis that PGHS-2 expression is regulated by mitogen activated protein kinases (MAPKs), and that over expression of PGHS-2 has anti-apoptotic effects mediated through differential gene expression.
Three Specific Aims have been outlined.
Specific Aim 1 will evaluate the hypothesis that activation of MAPKs represents the convergence point of divergent stimuli of PGHS-2 expression, and this is negatively regulated by dual-specificity phosphatases. Here, the effects on PGHS-2 expression after adenovirus mediated transfer of genes encoding wild type and mutant ERK, JNK and p38 MAPKs, and dual-specificity phosphatases will be studied in kidney cells.
Specific Aim 2 will examine the hypothesis that PGHS-2 expression leads to anti-apoptotic effects in several cell types. Here the effect of adenovirus-mediated transfer of PGHS-2 gene into primary cells on apoptotic parameters will be studied. The mechanism of the anti-apoptotic effect will be studied by identifying genes controlled by PGHS-2 expression using cDNA micro-arrays.
In Specific Aim 3, the in vivo relevance of PGHS-2 expression and relation to the MAPK cascade will be evaluated using two rat models of glomerulonephritis (GN). Here, PGHS-2 expression and prostaglandin production will be assessed in rat glomeruli and correlated with MAPK activation and the histopathology of GN at defined time points of the disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK041684-12
Application #
6177267
Study Section
Pathology A Study Section (PTHA)
Program Officer
Scherbenske, M James
Project Start
1989-07-01
Project End
2003-06-30
Budget Start
2000-07-01
Budget End
2001-06-30
Support Year
12
Fiscal Year
2000
Total Cost
$235,836
Indirect Cost
Name
Medical College of Wisconsin
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
073134603
City
Milwaukee
State
WI
Country
United States
Zip Code
53226
Alexanian, Anna; Miller, Bradley; Roman, Richard J et al. (2012) 20-HETE-producing enzymes are up-regulated in human cancers. Cancer Genomics Proteomics 9:163-9
Sorokin, Andrey (2011) Endothelin signaling and actions in the renal mesangium. Contrib Nephrol 172:50-62
Yang, Chen; Sorokin, Andrey (2011) Upregulation of fibronectin expression by COX-2 is mediated by interaction with ELMO1. Cell Signal 23:99-104
Rufanova, Victoriya A; Alexanian, Anna; Ostendorf, Tammo et al. (2010) Endothelin signaling via guanine exchange factor C3G in renal glomerular mesangial cells. Can J Physiol Pharmacol 88:808-16
Chahdi, Ahmed; Sorokin, Andrey (2010) The role of beta(1)Pix/caveolin-1 interaction in endothelin signaling through Galpha subunits. Biochem Biophys Res Commun 391:1330-5
Pavlov, Tengis S; Chahdi, Ahmed; Ilatovskaya, Daria V et al. (2010) Endothelin-1 inhibits the epithelial Na+ channel through betaPix/14-3-3/Nedd4-2. J Am Soc Nephrol 21:833-43
Rufanova, Victoriya A; Alexanian, Anna; Wakatsuki, Tetsuro et al. (2009) Pyk2 mediates endothelin-1 signaling via p130Cas/BCAR3 cascade and regulates human glomerular mesangial cell adhesion and spreading. J Cell Physiol 219:45-56
Moriyama, Takahito; Sorokin, Andrey (2009) BK virus (BKV): infection, propagation, quantitation, purification, labeling, and analysis of cell entry. Curr Protoc Cell Biol Chapter 26:Unit 26.2
Alexanian, Anna; Rufanova, Victoriya A; Miller, Bradley et al. (2009) Down-regulation of 20-HETE synthesis and signaling inhibits renal adenocarcinoma cell proliferation and tumor growth. Anticancer Res 29:3819-24
Rufanova, Victoriya A; Lianos, Elias; Alexanian, Anna et al. (2009) C3G overexpression in glomerular epithelial cells during anti-GBM-induced glomerulonephritis. Kidney Int 75:31-40

Showing the most recent 10 out of 57 publications