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Title: Alternative splicing of fibronectin and effect of EDA+FN spliced variant on human podocyte in culture
Author: Madne, Tarunkumar H.
ISNI:       0000 0004 6057 6757
Awarding Body: St George's, University of London
Current Institution: St George's, University of London
Date of Award: 2017
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Chronic kidney disease (CKD) is one of the major global health problems which may result in organ dysfunction. Podocytes are specialised cell type which plays a critical role in filtration. The interaction and attachment of podocytes to the Glomerular basement membrane (GBM) is critical for maintenance and functioning of filtration barrier. GBM is dynamic in nature and contains different proteins. Healthy maintenance of GBM is essential for filtration. Fibronectin (Fn) is identified as one of the components of the GBM. Fn is an established marker of fibrosis and involved in cellular functions such as adhesion, migration, growth, differentiation, signalling etc. EDA+Fn (Extra domain A) is documented as pathological alternative spliced variant of Fn. In our studies, we are studying the podocyte production, splicing and interaction of EDA+Fn in physiology and pathology. I have investigated the TGFpi mediated alternative splicing of EDA+Fn and underlying signalling mechanism in human podocytes culture and effect of EDA+Fn on podocytes phenotype and responses. Experiments were conducted on transformed human podocytes. Human recombinant TGFpi was used as a primary agonist. Fn spliced variants were measured by RT-PCR, Western blotting (WB) and Immunofluorescence (IF). Underlying intracellular signalling pathways were studied by using specific chemical inhibitors. The biological activity of EDA+Fn was assessed by TLR4 active HEK Blue cells. Effects of Fn spliced variants on podocyte phenotype and responses were studied by growing the podocytes on EDA+Fn and EDA-Fn coated dishes. TGFpi induces the production and alternative splicing of Fn in human podocytes which is regulated by PI3K/Akt and P38 MAP kinase signalling pathways. EDA+Fn was found to be biologically active as it activates the TLR4 pathway. EDA+Fn significantly alters the podocyte phenotype and responses. Targeting alternative splicing of Fn in human podocyte could be the influential remedial approach for the renal diseases.
Supervisor: Not available Sponsor: Southwest Thames Institute for Renal Research, St Helier Hospital
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available