|Title:||The Role of miR-200b in Renal Fibrosis and Its in vivo Delivery Systems|
|Authors:||Tang, Owen Miao Yi|
|Publisher:||University of Sydney.|
Kolling Institute of Medical Research.
Northern Clinical School.
|Abstract:||Irreversible renal fibrosis represents the final common pathway of almost all chronic renal diseases and ultimately leads to end stage renal failure. Excessive accumulation of extracellular matrix proteins distinctively characterises this fibrotic process, which epithelial to mesenchymal transition (EMT) is thought to play a major role in. This thesis explored the role of miR-200b in preventing renal fibrosis with the focus of its role in EMT. miR-200b was found to be an important player in maintaining the characteristics of epithelial cells by regulating transcription factors Zeb1 and Sip1 in proximal tubular cells. Furthermore, a direct targeting and degradation mechanism of fibronectin mRNA by miR-200b was identified. Thus, miR-200b is shown to be a master regulator of EMT and over-expression of this miRNA may have therapeutic implication in patients with chronic renal diseases. Followed by the discovery of molecular mechanisms underlying the protective effects of miR-200b in renal fibrosis, the focus of the study was shifted to the various delivery methods aiming targeted expression of miR-200b in renal proximal tubules in vivo. The efficacy of gene delivery methods such as lentivirus mediated gene transfer, hydrodynamic injection, ultrasound microbubble mediated gene transfer and nanoparticles mediated gene uptake were investigated. Expression of the transgene was successfully detected in the kidney in lentivirus and ultrasound microbubble mediated gene transfer. Optimisation of each method is still in process aiming higher transfer efficiency. In the last chapter, the role of novel protein target PMEPA1 regulated by both miR-200b and TGFβ was also investigated using in vitro fibrosis model. However, both over-expression and loss of function studies demonstrated no significant changes in the expression level of E-Cadherin, fibronectin and collagen IV suggesting its involvement in pathways other than EMT.|
|Type of Work:||PhD Doctorate|
|Type of Publication:||Doctor of Philosophy Ph.D.|
|Appears in Collections:||Sydney Digital Theses (Open Access)|
|TANG Owen - Final thesis.pdf||Thesis||4.53 MB||Adobe PDF|
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