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Title: The biological role of factor-inhibiting hypoxia-inducible factor
Author: Khan, Moddasar N.
ISNI:       0000 0004 2687 0207
Awarding Body: Imperial College London
Current Institution: Imperial College London
Date of Award: 2009
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Factor Inhibiting Hypoxia-inducible Factor (FIH) is an asparaginyl hydroxylasewhich regulates the transcription factor Hypoxia-Inducible Factor (HIF), viahydroxylation of a conserved asparagine residue of the HIF-alpha subunits (twoisoforms of which are well established, HIF-1 and HIF-2alpha - HIF-3alpha isless well characterized currently). Other targets of FIH have also been reported. Little is known about both FIH expression and function. This thesis willinvestigate the expression of FIH in rodents, in cell lines and in renal cancertissue samples. In addition, RNAi technology was used to study FIH function.Clear cell renal cell carcinoma (CCRCC) is commonly associated withinactivation of tumour suppressor von-Hippel Lindau protein (VHL) andconstitutive activation of HIF. The main question I have addressed in this thesisis whether FIH decreases HIF activation in this setting. To address this I inhibitedFIH using several approaches. Specifically, using hypoxia, dimethyloxalylglycine(DMOG) and RNA interference (RNAi). Each of these increased the expressionof HIF target genes in two different CCRCC cell lines, RCC10 and RCC4. Investigating three different CCRCC cell lines, FIH inhibition decreased numbers of RCC4 and RCC10 cells growing in culture, which is likely to be due to an increase in expression of pro-apoptotic, FIH-regulated HIF target genes. Interestingly, 786-O cells exclusively express the HIF-2alpha isoform. Attenuation of FIH in this setting did not affect expression of HIF target genes, nor affect growth in culture. To determine if this was due to lack of FIH or may be due tolack of HIF-1alpha, I introduced HIF-1alpha via a viral vector. Following this, sensitivity to FIH was clearly demonstrated. This implies either specific 'protection' of HIF-2alpha in 786-O or more general FIH selectivity for the HIF-1alpha isoform. My findings contrast with two reports suggesting that FIH expression is suppressed in CCRCC. My findings give insight into how FIH asparagine hydroxylation regulates HIF, in particular in renal cancer and makes this enzyme a potential target for therapeutic inhibition, in the majority of renal cancers.
Supervisor: Maxwell, Patrick ; Kiriakidis, Serafim Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral