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Title: Drug target development and analysis of genome stability in cancer cells lacking the BAF180 subunit of the PBAF remodelling complex
Author: Hopkins, Suzanna
ISNI:       0000 0004 6350 1814
Awarding Body: University of Sussex
Current Institution: University of Sussex
Date of Award: 2017
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In eukaryotes, DNA is packaged into a highly condensed structure, known as chromatin. Several complexes facilitate the remodelling of chromatin, for example, INO80, NURD and SWI/SNF, which attach to tightly bound chromatin, allowing its relaxation by nucleosome sliding, unwrapping, histone eviction and exchange of histone variants. The activities carried out by these chromatin remodelling complexes are thought to be integral in the prevention of cancer cell formation. Recently, whole exome sequencing has identified frequent mutations in subunits of the SWI/SNF chromatin remodelling complex, at a frequency that rivals p53. Strikingly, the BAF180 (PBRM1) subunit of the PBAF variant of SWI/SNF remodelers is mutated in over 40% of clear cell renal cell carcinoma (ccRCC), a cancer with typically poor prognosis and limited treatment options to date. This work embodies four main results chapters that aim to identify novel synthetic lethal gene candidates with BAF180, with a view to targeting these gene candidates with chemotherapeutic drugs. In the first chapter we work through a short list of hypothesis driven potential synthetic lethal candidates and identify the genes KAT2A, RNF4, EZH2 and BAP1 as potential synthetic lethal partners for BAF180. Chapter two describes the development of both stable shRNA and CRISPR/Cas9-derived BAF180-deficient cell lines that were used both in this study as well as for other ongoing projects. The third chapter outlines the set-up of a high-throughput synthetic lethal siRNA (HTS) screen and determines potential synthetic lethal interactions identified here. The final chapter examines various PARP genes, identified as hits in HTS screening, to further explore the interaction between PARP and BAF180. We find that PARP1 and PARP3 are synthetic lethal with BAF180 and treatment with various siRNA's and PARP inhibitors in BAF180 deficient mammalian cells results in specific cell death. A phenotype that could be clinically exploited for treatment of ccRCC.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: QD0415 Biochemistry