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Title: A study of the effects of Ki-Ras inhibition by antisense deoxyoligonucleotides in a rat model of renal fibrosis
Author: Wang, Joe
Awarding Body: King's College London (University of London)
Current Institution: King's College London (University of London)
Date of Award: 2012
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Ras monomeric GTPases are key molecules in the signalling pathway of renal fibrogenesis and exist in three isoforms (Kirsten, Harvey and Neural). The aim of this thesis was to determine whether inhibition of Ki-Ras expression by ASOs could ameliorate the process of renal fibrosis. All isoforms of Ras were expressed in rat renal fibroblasts (primary culture and cell line (NRK-49F)) and in a rat epithelial cell line (NRK-52E). In vitro administration of Ki-Ras ASOs resulted in a specific knockdown of the isoform mRNA in all cells. An associated rise in Ha-Ras mRNA was noted in fibroblast cells but absent in epithelial cells. ASOs administration in non-diseased male Wistar rats resulted in specific renal Ki- Ras mRNA knockdown and an associated decrease in Ras protein expression. Oligo accumulation was seen in the renal proximal tubular cells with extra-renal deposition seen in the liver, heart and wound-tissue. Significant hepatic Ki-Ras mRNA knockdown and inflammation was also demonstrated in ASO treated subjects. In a rat model of unilateral ureteric obstruction (UUO) a significant rise in both Ki- Ras and N-Ras mRNA expression in the obstructed kidneys compared to nonobstructed kidney samples was demonstrated. Ki-Ras ASOs administration to models of UUO significantly reduced the isoform mRNA expression, when compared to vehicle-only and scrambled-oligos (SO), and resulted in a significant decrease in renal fibrosis scores, based on trichrome and picrosirius red staining, and interstitial collagen I and !-SMA expression, based on immunohistological staining and western blotting. ASO administration resulted in an increase in interstitial cell expression of FSP-1, Ki- 67 and ED-1. These findings were absent in vehicle-only and control oligo groups. In summary, administration of ASOs specifically and significantly reduce Ki-Ras mRNA expression and ameliorate fibrosis in a rat model of renal fibrosis in the presence of increased interstitial cell proliferation and inflammation.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available