The long term goal of our research proposal is to define in vitro and in vivo the membrane, cytoplasmic and nuclear signal transduction mechanisms by which cytokines activate mesangial and other glomerular cells. Expression of growth factors and immune-modulatory cytokines and their receptors is upregulated in renal tissue of patients and experimental animals with proliferative glomerular diseases. This proposal will explore the role of tyrosine phosphorylation/dephosphorylation mechanisms in activating mesangial cells. Our hypothesis is that the sustained activation of certain signals is responsible for changes in behavior of glomerular cells during disease. Thus, understanding the regulation of cell signaling and gene expression transduced by activated cytokine receptors provides the best chance to identify the most relevant targets for therapeutic intervention. We propose to carry out in vivo and in vitro studies to define mechanisms and pathways that platelet-derived growth factor (PDGF) and immune-modulary cytokines such as interferon- gamma, interleukin-1 (IL-1), IL-6 and tumor necrosis factor utilize to elicit their biologic effects in cultured mesangial cells and in experimental proliferative glomerulonephritis. Signals to be studied include tyrosine phosphorylation of membrane and cytoplasmic proteins that associate with the PDGF receptor or non-receptor tyrosine kinases stimulated by other cytokines. Signals to be explored include phosphatidyl inositol 3 kinase Janus kinases (JAKs). Immunoprecipitation and immunoblotting techniques will be used to identify and characterize tyrosine-phosphorylated proteins. Cytoplasmic and nuclear transcription factors that recognize specific DNA sequences present on target genes will be identified by gel retardation using labeled specific oligonucleotides. Activation of cytoplasmic transcription factors including members of the signal transducers and activators of transcription (STATS) will be explored. To define the mechanisms of cytokine interaction, we will periodically examine the effect of selected relevant cytokines interaction in glomerular cells as well as after in vivo administration. To deactivate signals, gene therapy approach will be utilized to deliver tyrosine phosphatases to the kidney using retroviral vectors. These studies should define signals involved in mediating glomerular pathology and establish effective therapeutic modalities to treat inflammatory forms of glomerulonephritis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK043988-08
Application #
2900245
Study Section
Pathology A Study Section (PTHA)
Program Officer
Hirschman, Gladys H
Project Start
1991-05-15
Project End
2001-03-31
Budget Start
1999-04-01
Budget End
2000-03-31
Support Year
8
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Texas Health Science Center San Antonio
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
800772162
City
San Antonio
State
TX
Country
United States
Zip Code
78229
Block, Karen; Gorin, Yves; Abboud, Hanna E (2009) Subcellular localization of Nox4 and regulation in diabetes. Proc Natl Acad Sci U S A 106:14385-90
Block, Karen; Ricono, Jill M; Lee, Duck-Yoon et al. (2006) Arachidonic acid-dependent activation of a p22(phox)-based NAD(P)H oxidase mediates angiotensin II-induced mesangial cell protein synthesis and fibronectin expression via Akt/PKB. Antioxid Redox Signal 8:1497-508
Kim, Nam-Ho; Rincon-Choles, Hernan; Bhandari, Basant et al. (2006) Redox dependence of glomerular epithelial cell hypertrophy in response to glucose. Am J Physiol Renal Physiol 290:F741-51
Gorin, Yves; Block, Karen; Hernandez, James et al. (2005) Nox4 NAD(P)H oxidase mediates hypertrophy and fibronectin expression in the diabetic kidney. J Biol Chem 280:39616-26
Gorin, Yves; Ricono, Jill M; Wagner, Brent et al. (2004) Angiotensin II-induced ERK1/ERK2 activation and protein synthesis are redox-dependent in glomerular mesangial cells. Biochem J 381:231-9
Ghosh Choudhury, Goutam; Abboud, Hanna E (2004) Tyrosine phosphorylation-dependent PI 3 kinase/Akt signal transduction regulates TGFbeta-induced fibronectin expression in mesangial cells. Cell Signal 16:31-41
Gorin, Yves; Ricono, Jill M; Kim, Nam-Ho et al. (2003) Nox4 mediates angiotensin II-induced activation of Akt/protein kinase B in mesangial cells. Am J Physiol Renal Physiol 285:F219-29
Simon, Matthias; Feliers, Denis; Arar, Mazen et al. (2002) Cloning of the 5'-flanking region of the murine bone morphogenetic protein-7 gene. Mol Cell Biochem 233:31-7
Gorin, Y; Kim, N H; Feliers, D et al. (2001) Angiotensin II activates Akt/protein kinase B by an arachidonic acid/redox-dependent pathway and independent of phosphoinositide 3-kinase. FASEB J 15:1909-20
Choudhury, G G; Grandaliano, G; Jin, D C et al. (2000) Activation of PLC and PI 3 kinase by PDGF receptor alpha is not sufficient for mitogenesis and migration in mesangial cells. Kidney Int 57:908-17

Showing the most recent 10 out of 45 publications