Functional Proteome analysis of the molecular mechanism of (myo)fibroblast differentiation
Research output: Thesis › Doctoral Thesis (compilation)
Tissue remodeling is observed in several physiological conditions such as embryogenesis, senescence, wound healing and pregnancy, as well as in several pathological conditions such as fibrosis and tumorgenesis. Fibroblasts play a key role in this tissue remodeling due to their capability of altering the turnover of the extracellular matrix. During the remodeling process, the fibroblasts are believed to differentiate into fibroblast-smooth muscle-like cells called myofibroblasts. This differentiation process is mediated by a well-orchestrated interplay between the extracellular matrix and growth factors such as platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-beta ). In this thesis, the overall aim was to investigate the mechanism for fibroblast activation by employing various proteomic technologies. We present a mechanistic explanation as to how TGF-beta induces alternative splicing of a number of extracellular matrix components that have been found to be essential for the differentiation process. This thesis also shows how specific carbohydrate moieties can modulate the PDGF-induced cellular response. By isolating fibroblast populations from different bronchial locations, the in vitro results could be compared with the bronchial-derived fibroblast populations by proteome profiling. To achieve these goals, application of a number of proteomic tools was necessary such as two-dimensional gel electrophoresis, multidimensional chromatography and mass spectrometers utilizing both laser desorption and electrospray ionization. A database was also constructed to store and analyze the data derived from the studies. In summary, the results showed that myofibroblast differentiation was dependent upon a coordinate interplay between the extracellular matrix components and growth factors. These growth factors act locally on the fibroblasts to induce a tissue remodeling similar to that observed in diseases such as asthma.
|Research areas and keywords||
Subject classification (UKÄ) – MANDATORY
|Award date||2003 Oct 3|
|Publication status||Published - 2003|
Defence details Date: 2003-10-03 Time: 09:15 Place: GK salen, BMC External reviewer(s) Name: Andrén, Per Title: Dr Affiliation: Biological and Medical Mass Spectrometry Research Laboratory, Department of Pharmaceutical Biosciences, Uppsala. --- Article: I. Johan Malmström, Kristoffer Larsen, Lennart Hansson, Cleas-Göran Löfdahl, Ole Nöregaard-Jensen, György Marko-Varga and Gunilla Westergren-Thorsson. Proteoglycan and Proteome profiling of Central Human Pulmonary Fibrotic Tissue Utilizing Miniaturized Sample Preparation: A Feasibility study. Proteomics. Apr: 2(4):394-404 (2002). Article: II. Johan Malmström, Ellen Tufvesson, Claes-Göran Löfdahl, Lennart Hansson, György Marko-Varga and Gunilla Westergren-Thorsson. Activation of Platelet-Derived Growth Factor Pathway in Human Asthmatic Pulmonary-Derived Mesenchymal Cells. Electrophoresis Jan 24 (1-2) 276-85 (2003). Article: III. Johan Malmström and Gunilla Westergren-Thorsson. Heparan sulphate upregulates platelet-derived growth factor receptors on human lung fibroblasts. Glycobiology. 8 (12), 1149-1156. (1998). Article: IV. Ellen Tufvesson, Johan Malmström, György Marko-Varga and Gunilla Westergren-Thorsson. Biglycan isoforms with differences in polysaccharide substitution and core protein in human lung fibroblasts. European Journal of Biochemistry, 269, 3688-3696 (2002). Article: V. Johan Malmström, Kristoffer Larsen, Lars Malmström, Ellen Tufvesson, Ken Parker, Jason Marchese, Brian Williamson, Dale Patterson, Steve Martin, Peter Juhasz, Gunilla Westergren-Thorsson and György Marko-Varga. Nano-Capillary Liquid Chromatography Interfaced to Tandem MALDI and ESI mass Spectrometry: Mapping the nuclear proteome of human fibroblasts. In press Electrophoresis (2003). Article: VI. Johan Malmström, Kristoffer SA Larsen, Lars Malmström, Ellen Tufvesson, Ken C Parker, Jason Marchese, Brian L Williamson, Steve J Hattan, Dale H Patterson, Steve A Martin, Armin Graber, Peter Juhasz, György Marko-Varga and Gunilla Westergren-Thorsson. Quantitative proteomic analysis of Transforming growth factor stimulated human lung fibroblasts: Serine and arginine-rich splicing factors as a nuclear targets for TGF-beta dependent induction of alternative splicing. Submitted to Journal of Biological Chemistry.