The signaling mechanisms underlying kidney fibrosis continue to be of significant concern and interest. This proposal presents a strategy for extending our previous studies that examined how TGF-2 stimulates accumulation of type I collagen in a cellular model of renal fibrogenesis. In the previous grant period, we demonstrated that numerous signaling pathways interact with TGF-2/Smad signaling, focusing particularly on the role of the extracellular signal-regulated kinase (ERK) MAP kinase. We determined that the Smad3 and ERK pathways together promote TGF-2-stimulated collagen expression in renal mesangial and tubular epithelial cells. ERK serves a critical role in supporting Smad3-mediated collagen gene transcription, but does not activate the COL1A2 promoter in the absence of Smad activation;it is thus necessary but not sufficient for collagen expression. ERK phosphorylates serines and threonines in the linker region (LR) domain of Smad3. Additional Preliminary Studies indicate that blocking 1v23 integrin or PI3-kinase activity inhibits ERK activation and the collagen response. For this renewal, we propose the HYPOTHESIS that an adhesion-dependent signaling pathway from 1v23-integrin to ERK promotes renal cell type I collagen expression through phosphorylation of linker-region serine and/or threonine residues of Smad3;and PI3K, which acts at several points along the pathway leading to ERK, is a promising target for blunting the collagen response to TGF-2 both in vitro and in vivo. To test this hypothesis, we will pursue the following AIMS: First, we will determine the molecular mechanisms by which 1v23 integrin promotes TGF-2-stimulated renal cell fibrogenesis, establishing the roles of Rac1 and PI3K in this process. Second, we will determine the mechanism(s) through which ERK and Smad3 synergize to promote TGF-2-stimulated collagen expression by examining how specific phosphorylation events affect Smad3 signaling of collagen I expression and which of these phosphorylations are dependent upon PI3K and/or ERK. Third, we will determine how 1v23, PI3K and ERK interact to modulate the Smad3 LR in the renal fibrosis models of adriamycin nephropathy and selective podocyte ablation. These studies should provide new insights into integrin regulation of Smad signaling, address unsettled issues regarding the role of the Smad3 LR, and offer potential approaches to interrupting the fibrogenic process in kidneys. SIGNIFICANCE: Despite extensive research, little is known about how TGF-2 signaling is regulated to cause renal fibrosis. Our studies of non-canonical signaling pathways and the role of Smad3-LR will address key issues regarding the specificity of signals in fibrogenesis. By elucidating the signaling events underlying kidney fibrosis we hope to better understand fibrogenic mechanisms, how to interrupt them and, ultimately, how to treat them.
Chronic, progressive kidney disease, which costs the nation many billions of dollars in health care each year, involves the accumulation of scar tissue components such as type I collagen. Transforming growth factor (TGF)-2 is a protein that plays a central role in scarring of the kidney. Although we have made much progress in understanding how TGF-2 works, we have not learned how to regulate its actions in order to prevent or minimize kidney scarring. This proposal requests renewal of support for a project presently in its 14th year, seeking to understand how TGF-2 stimulates synthesis of type I collagen. By defining the molecular events inside the cell that lead to scarring, we hope to identify targets to prevent such scarring and decrease the incidence of chronic kidney failure.
|Schnaper, H William (2014) Remnant nephron physiology and the progression of chronic kidney disease. Pediatr Nephrol 29:193-202|
|Liu, Xiaoying; Hubchak, Susan C; Browne, James A et al. (2014) Epidermal growth factor inhibits transforming growth factor-?-induced fibrogenic differentiation marker expression through ERK activation. Cell Signal 26:2276-83|
|Hanna, Christian; Hubchak, Susan C; Liang, Xiaoyan et al. (2013) Hypoxia-inducible factor-2* and TGF-* signaling interact to promote normoxic glomerular fibrogenesis. Am J Physiol Renal Physiol 305:F1323-31|
|Chen, Guang; Chen, Xing; Sukumar, Aravin et al. (2013) TGF* receptor I transactivation mediates stretch-induced Pak1 activation and CTGF upregulation in mesangial cells. J Cell Sci 126:3697-712|
|Browne, J A; Liu, X; Schnaper, H W et al. (2013) Serine-204 in the linker region of Smad3 mediates the collagen-I response to TGF-* in a cell phenotype-specific manner. Exp Cell Res 319:2928-37|
|Hodgkins, Kavita S; Schnaper, H William (2012) Tubulointerstitial injury and the progression of chronic kidney disease. Pediatr Nephrol 27:901-9|
|Finer, Gal; Schnaper, H William; Kanwar, Yashpal S et al. (2012) Divergent roles of Smad3 and PI3-kinase in murine adriamycin nephropathy indicate distinct mechanisms of proteinuria and fibrogenesis. Kidney Int 82:525-36|
|Basu, Rajit K; Hubchak, Susan; Hayashida, Tomoko et al. (2011) Interdependence of HIF-1ýý and TGF-ýý/Smad3 signaling in normoxic and hypoxic renal epithelial cell collagen expression. Am J Physiol Renal Physiol 300:F898-905|
|Schnaper, H William; Hubchak, Susan C; Runyan, Constance E et al. (2010) A conceptual framework for the molecular pathogenesis of progressive kidney disease. Pediatr Nephrol 25:2223-30|
|Schnaper, H William; Jandeska, Sara; Runyan, Constance E et al. (2009) TGF-beta signal transduction in chronic kidney disease. Front Biosci 14:2448-65|
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