CELLULAR REGULATION OF PROSTAGLANDIN SYNTHESIS

Summary

Principal Investigator: A Sorokin
Abstract: Renal glomerular mesangial cells (GMC) are important participants in glomerular injury. GMC apoptosis is the major mechanism for resolution of glomerular hypercellularity in experimental mesangial proliferative nephritis. Resistance of GMC to apoptosis may account for progression of injury to end stage disease since cell proliferation is not counteracted by extra cells deletion. The long term objectives of this application are to elucidate mechanisms of resistance of GMC to apoptosis. This proposal will examine the hypothesis that cyclooxygenase 2 (Cox-2), which generate prostaglandins from arachidonic acid, protects GMC from apoptosis via up-regulation of genes, that inhibit activation activity of nitric oxide synthases (NOS) and regulate efflux preliminary data indicating that 1) Cox-2 inhibits apoptosis of pro-apoptotic Bcl-2 family proteins, control of xenobiotics. The hypothesis is based on our in several types of cells including GMC;2) Cox-2 up-regulates expression of Protein Inhibitor of NOS (PIN), and multidrug resistance P-glycoprotein (MDR1);3) apoptotic agents increase cellular NO production. The Bim-dependent Bax activation will be measured by immunofluorescence. The activity of NOS and generation of NO will be evaluated by measurement of arginine to citrullin conversion and by quantification of NO oxidation products with Sievers analyzer in GMC induced to apoptosis in the presence and absence of Cox-2 expression. We will identify products of cyclooxygenase activity in isolated glomeruli and in GMC infected with recombinant adenovirus encoding Cox-2 by Mass Spectrometer coupled with Liquid Chromatography (LC/MS) in the presence and absence of selective Cox-2 inhibitors. To evaluate the role of cyclooxygenase activity in the regulation of gene expression and anti-apoptotic action of Cox-2 we will test the ability of Cox-2 products (identified by LC/MS) to mimic Cox-2 effects and will evaluate the anti-apoptotic properties of Cox-2 mutants which lack the cyclooxygenase activity, but retain peroxidase activity. We will also investigate the effects of blocking the Cox-2 activity on progression of proliferative glomerulonephritis, hypertension and diabetic nephropathy in in vivo experimental models. The significance of this study is that it will provide new insights into the response of GMC to injury and help to understand why elimination of proliferating GMC by apoptosis in the course of progressive renal injury is often inefficient.
Funding Period: 1989-07-01 - 2010-11-30
more information: NIH RePORT

Top Publications

  1. pmc 20-HETE-producing enzymes are up-regulated in human cancers
    Anna Alexanian
    Kidney Disease Center, Department of Medicine and Physiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    Cancer Genomics Proteomics 9:163-9. 2012
  2. ncbi Endothelin signaling and actions in the renal mesangium
    Andrey Sorokin
    Kidney Disease Center, Division of Nephrology, Department of Medicine, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
    Contrib Nephrol 172:50-62. 2011
  3. pmc Upregulation of fibronectin expression by COX-2 is mediated by interaction with ELMO1
    Chen Yang
    Division of Nephrology and Kidney Disease Center, Department of Medicine, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA
    Cell Signal 23:99-104. 2011
  4. ncbi Endothelin signaling via guanine exchange factor C3G in renal glomerular mesangial cells
    Victoriya A Rufanova
    Department of Medicine, Division of Nephrology and Kidney Disease Center, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA
    Can J Physiol Pharmacol 88:808-16. 2010
  5. pmc Endothelin-1 inhibits the epithelial Na+ channel through betaPix/14-3-3/Nedd4-2
    Tengis S Pavlov
    Department of Physiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    J Am Soc Nephrol 21:833-43. 2010
  6. pmc The role of beta(1)Pix/caveolin-1 interaction in endothelin signaling through Galpha subunits
    Ahmed Chahdi
    Division of Nephrology and Kidney Disease Center, Department of Medicine, Medical College of Wisconsin, Milwaukee, WI 53226, United States
    Biochem Biophys Res Commun 391:1330-5. 2010
  7. pmc Down-regulation of 20-HETE synthesis and signaling inhibits renal adenocarcinoma cell proliferation and tumor growth
    Anna Alexanian
    Kidney Disease Center, Department of Medicine and Physiology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA
    Anticancer Res 29:3819-24. 2009
  8. pmc BK virus (BKV): infection, propagation, quantitation, purification, labeling, and analysis of cell entry
    Takahito Moriyama
    Medical College of Wisconsin, Milwaukee, Wisconsin, USA
    Curr Protoc Cell Biol . 2009
  9. pmc Pyk2 mediates endothelin-1 signaling via p130Cas/BCAR3 cascade and regulates human glomerular mesangial cell adhesion and spreading
    Victoriya A Rufanova
    Division of Nephrology, Department of Medicine, Kidney Disease Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA
    J Cell Physiol 219:45-56. 2009
  10. pmc C3G overexpression in glomerular epithelial cells during anti-GBM-induced glomerulonephritis
    Victoriya A Rufanova
    Division of Nephrology and Kidney Disease Center, Department of Medicine, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
    Kidney Int 75:31-40. 2009

Scientific Experts

  • A Sorokin
  • Victoriya A Rufanova
  • Anna Alexanian
  • Ahmed Chahdi
  • Bradley Miller
  • Takahito Moriyama
  • Chen Yang
  • Richard J Roman
  • Tengis S Pavlov
  • Jacob Huh
  • Vladislav Levchenko
  • Tammo Ostendorf
  • Alexander Staruschenko
  • Dirk Bokemeyer
  • Simon Prosser
  • Oleh Pochynyuk
  • Alain Vandewalle
  • Daria V Ilatovskaya
  • Adam Lerner
  • Ann McGinty
  • Elena Sorokina
  • Averia Flasch
  • Elias Lianos
  • Tetsuro Wakatsuki
  • Mukut Sharma
  • Andrejs Liepins
  • Christopher Andrekopoulos
  • Balaraman Kalyanaraman
  • Vimal A Patel
  • Jacek Zielonka

Detail Information

Publications16

  1. pmc 20-HETE-producing enzymes are up-regulated in human cancers
    Anna Alexanian
    Kidney Disease Center, Department of Medicine and Physiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    Cancer Genomics Proteomics 9:163-9. 2012
    ....
  2. ncbi Endothelin signaling and actions in the renal mesangium
    Andrey Sorokin
    Kidney Disease Center, Division of Nephrology, Department of Medicine, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
    Contrib Nephrol 172:50-62. 2011
    ..Plentiful evidence suggests an essential role of ET-1-signaling and action in the renal mesangium for renal biology and pathobiology...
  3. pmc Upregulation of fibronectin expression by COX-2 is mediated by interaction with ELMO1
    Chen Yang
    Division of Nephrology and Kidney Disease Center, Department of Medicine, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA
    Cell Signal 23:99-104. 2011
    ..The interaction of ELMO1 with COX-2 could play an important role in the development and progression of renal glomerular injury...
  4. ncbi Endothelin signaling via guanine exchange factor C3G in renal glomerular mesangial cells
    Victoriya A Rufanova
    Department of Medicine, Division of Nephrology and Kidney Disease Center, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA
    Can J Physiol Pharmacol 88:808-16. 2010
    ....
  5. pmc Endothelin-1 inhibits the epithelial Na+ channel through betaPix/14-3-3/Nedd4-2
    Tengis S Pavlov
    Department of Physiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    J Am Soc Nephrol 21:833-43. 2010
    ..Taken together, these results suggest that the inhibitory effects of chronic ET-1 on ENaC result from betaPix interacting with the 14-3-3/Nedd4-2 pathway...
  6. pmc The role of beta(1)Pix/caveolin-1 interaction in endothelin signaling through Galpha subunits
    Ahmed Chahdi
    Division of Nephrology and Kidney Disease Center, Department of Medicine, Medical College of Wisconsin, Milwaukee, WI 53226, United States
    Biochem Biophys Res Commun 391:1330-5. 2010
    ....
  7. pmc Down-regulation of 20-HETE synthesis and signaling inhibits renal adenocarcinoma cell proliferation and tumor growth
    Anna Alexanian
    Kidney Disease Center, Department of Medicine and Physiology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA
    Anticancer Res 29:3819-24. 2009
    ..We examined the ability of inhibitors of the synthesis or actions of 20-HETE, metabolite of arachidonic acid, to inhibit proliferation of human renal carcinoma cell lines...
  8. pmc BK virus (BKV): infection, propagation, quantitation, purification, labeling, and analysis of cell entry
    Takahito Moriyama
    Medical College of Wisconsin, Milwaukee, Wisconsin, USA
    Curr Protoc Cell Biol . 2009
    ..This unit describes the handling of BKV, BKV propagation, determination of titer and ability to infect cells, as well as purification and labeling of BKV in order to analyze BKV cell entry...
  9. pmc Pyk2 mediates endothelin-1 signaling via p130Cas/BCAR3 cascade and regulates human glomerular mesangial cell adhesion and spreading
    Victoriya A Rufanova
    Division of Nephrology, Department of Medicine, Kidney Disease Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA
    J Cell Physiol 219:45-56. 2009
    ..Our data suggest that ET-1 stimulated the GTPase Rap1 (but neither RhoA nor Ras) by a mechanism involving Pyk2 activation and recruitment of the p130Cas/BCAR3 complex in GMC...
  10. pmc C3G overexpression in glomerular epithelial cells during anti-GBM-induced glomerulonephritis
    Victoriya A Rufanova
    Division of Nephrology and Kidney Disease Center, Department of Medicine, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
    Kidney Int 75:31-40. 2009
    ..We found that C3G was overexpressed in accelerated anti-GBM antibody-induced glomerulonephritis and suggest that this modulates glomerular epithelial cell morphology and behavior...
  11. pmc Repression of BK virus infection of human renal proximal tubular epithelial cells by pravastatin
    Takahito Moriyama
    Division of Nephrology and Kidney Disease Center, Department of Medicine, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    Transplantation 85:1311-7. 2008
    ....
  12. pmc Endothelin-1 couples betaPix to p66Shc: role of betaPix in cell proliferation through FOXO3a phosphorylation and p27kip1 down-regulation independently of Akt
    Ahmed Chahdi
    Kidney Disease Center, Department of Medicine, Division of Nephrology, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    Mol Biol Cell 19:2609-19. 2008
    ..Our results identify p66Shc and FOXO3a as novel partners of beta(1)Pix and represent the first direct evidence of beta(1)Pix in cell proliferation via Erk/p66Shc-dependent and Akt-independent mechanisms...
  13. ncbi CrkII associates with BCAR3 in response to endothelin-1 in human glomerular mesangial cells
    Victoriya A Rufanova
    Department of Medicine Kidney Disease Center, Division of Nephrology, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, Wisconsin 53226, USA
    Exp Biol Med (Maywood) 231:752-6. 2006
    ....
  14. ncbi Cyclooxygenase 2 rescues LNCaP prostate cancer cells from sanguinarine-induced apoptosis by a mechanism involving inhibition of nitric oxide synthase activity
    Jacob Huh
    Department of Medicine and Free Radical Research Center, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53266, USA
    Cancer Res 66:3726-36. 2006
    ....
  15. ncbi Cyclooxygenase-2 rescues rat mesangial cells from apoptosis induced by adriamycin via upregulation of multidrug resistance protein 1 (P-glycoprotein)
    Bradley Miller
    Department of Medicine, Cardiovascular Research Center, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA
    J Am Soc Nephrol 17:977-85. 2006
    ..These data suggest that COX-2 protects GMC from ADR-mediated apoptosis via transcriptional upregulation of MDR-1 and that induced COX-2 expression would lessen ADR nephrotoxicity...
  16. ncbi Endothelin 1 induces beta 1Pix translocation and Cdc42 activation via protein kinase A-dependent pathway
    Ahmed Chahdi
    Department of Medicine, Division of Nephrology, Kidney Disease Center and Cardiovascular Research Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA
    J Biol Chem 280:578-84. 2005
    ..Our results have provided evidence that stimulation of PKA pathway by ET-1 or cAMP analog results in beta(1)Pix phosphorylation, which in turn controls beta(1)Pix translocation to focal complexes and Cdc42 activation...