Oxford Wolfson Marriott Graduate Scholarship: Investigating changes in RNA-binding proteins in hypoxia as a means of increasing radiosensitivity
Primary Supervisor: Professor Ester Hammond
Second Supervisor: Professor Geoff Higgins
Project Overview
This funded project offers an Oxford Wolfson Marriott Graduate Scholarship, providing a living stipend at UKRI standard rate. Please note, the studentship covers fees at the UK Home rate only. International (Overseas fee status) students need to be aware that they would need to pay the shortfall between Home and International fees.
Hypoxia (the condition of insufficient oxygen) is a common feature of the tumour microenvironment leading to therapy resistance and poor patient prognosis. Hypoxic cells undergo transcriptional stress, which we have recently shown to involve R-loop accumulation, nucleolar reorganisation and repression of rRNA synthesis. Furthermore, our previous study demonstrated that increasing R-loop accumulation in hypoxia led to significant changes in expression of RNA binding proteins. These data led us to hypothesis that the study of RNA-binding proteins in the context of hypoxia-induced transcriptional stress could identify both novel biology and therapeutic targets. Therefore, we set out to identify novel RNA-binding and/or R-loop-binding proteins that regulate the transcriptional stress response. We successfully isolated RNA-binding proteins from cells and generated samples for mass spectrometry to quantify changes in the RNA-bound proteome between normoxic and hypoxic cells. Importantly, hypoxic samples were also prepared with cells overexpressing RNase H1, an endonuclease that degrades the RNA strand of R-loops, to investigate RNA-bound proteome differences between hypoxic cells with and without significant R-loop levels. Approximately 70 proteins were identified which showed changes in RNA binding activity in hypoxic conditions, of these 20 also changed in an RNase H1 dependent manner suggesting a role at R-loops. The aim of this project is to further define the roles of specific RNA-binding and/or R-loop-binding proteins in hypoxia and identify potential targets for novel therapeutic strategies.
Training Opportunities
The student can expect training in molecular/cell biology, biochemistry and proteomics. In addition, preclinical testing of promising candidates and understanding of the challenges to successful clinical translation. The student is expected to spend time in both the Hammond and Mardakheh labs, the latter in particular for biochemistry/proteomics training.
References
Ma, T.S., Worth, K.R., Maher, C., Ng, N., Beghè, C., Gromak, N., Rose, A.M. and Hammond, E.M., 2023. Hypoxia-induced transcriptional stress is mediated by ROS-induced R-loops. Nucleic Acids Research, 51(21), pp.11584-11599.