The major objective of this research program is to provide the applicant with new and intensive training in cell and molecular biology to expand her research tools with the purpose of becoming an independent investigator. To achieve this goal the applicant will train in the rich scientific environment of the University of Florida, under the sponsorship of Harry Nick, Ph. D. and co-sponsorship of Anupam Agarwal, M.D. The long-term goals of this proposal are to determine the cytoprotective actions and molecular mechanisms of heme oxygenase-1 gene expression in response to transforming growth factor-beta1 (TGF-beta1) in the kidney. Increased expression of TGF-beta1 in glomeruli and tubulointerstitium leads to increased cell proliferation and deposition of extracellular matrix, resulting in progressive kidney fibrosis and, ultimately, end stage renal disease. TGF-beta1, however, stabilizes and attenuates tissue injury through the activation of cytoprotective proteins. One such cytoprotective protein is heme oxygenase-1, which has antioxidant, anti-inflammatory and anti-apoptotic properties. Induction of heme oxygenase-1 occurs as an adaptive and beneficial response to renal injury.
Aims I and II of this proposal will evaluate the cellular effects of TGF-beta1-mediated heme oxygenase-1 induction on cell proliferation, deposition of extracellularmatrix, and apoptosis in the absence or presence of Smad proteins, molecules involved in TGF-beta1 signalling.
Aims III and IV will identify the transcriptional mechanisms involved in controlling TGF-beta1-mediated heme oxygenase-1 induction. Further understanding of the cytoprotective actions and molecular mechanisms of heme oxygenase-1 gene expression in response to TGF-beta1 will be important in designing interventional strategies in the pathogenesis of renal injury. The proposed studies will be the basis for training the applicant in state-of-the-art cell and molecular biology techniques such as recombinant DNA, transfection, northern and immunoblot analyses, promoter-deletion analysis, electrophoretic mobility shift assays and nuclear run on assays, which will greatly enhance the applicant?s research career development towards an independent researcher.
