Dr F Manson, Dr Lisa Swanton
Applications accepted all year round
Self-Funded PhD Students Only
About the Project
Up to half of human diseases are linked to protein misfolding. These include more than 15 retinal dystrophies caused by mutations in retinal pigment epithelium (RPE) proteins such as bestrophin-1. Building on our preliminary data showing the expression, localisation and function of mutant bestrophin-1 proteins can be rescued by small molecule chaperones and proteostasis regulators, we propose to extend this approach to other retinal diseases caused by misfolding of mutant RPE proteins. We will transfect RPE cell lines and RPE generated from induced pluripotent stem cells (iPSC-RPE) to study disease-causing mutant RPE proteins that have reported defects in trafficking, expression, and function. These include BEST1 (bestrophin-1), C1QTNF5, EFEMP1 (fibulin 3), FBLN5 (fibulin 5) and RDH5.
We will first define the folding, localisation and/or functional defects of the mutant proteins in order to provide cell models for the corresponding diseases. This will include immunofluorescence co-localisation with subcellular markers, cycloheximide chase experiments to study protein secretion, and assays for protein modification in the ER. Where appropriate, specific functional assays will be established. In the next phase of the project the fate of and cellular response to mutant RPE proteins will be determined. This will include assaying for autophagy, activation of the unfolded protein response pathway, and cell growth kinetics. Lastly, we will screen a range of chemical chaperones and proteostasis regulators to identify those that can restore the expression, localisation and function of the mutant proteins.
This strategy targets common pathways that can be manipulated to prevent disease development for a large number of diseases and is an important step towards in vivo testing and human clinical trials.
In addition to training opportunities provided by the academic and researcher development programme, specific training will be provided in a wide variety of transferrable laboratory skills. These include a wide range of molecular and cell biology techniques including PCR, cloning, immunoblotting, cell culture, immunofluorescent and confocal microscopy. Functional assays will be developed and will depend on the protein under study.
Candidates are expected to hold a minimum upper-second (or equivalent) undergraduate degree in a related biomedical/biological science. Previous laboratory experience is essential. Previous experience in molecular or cell biology, or a Masters qualification or relevant industrial laboratory experience, would be an advantage. International students must meet the faculty’s minimum entry requirements for postgraduate programmes.
This 3-year full-time PhD is open to candidates able to provide evidence of self-arranged funding/ sponsorship and is due to commence from January 2017 onwards.
Any enquiries relating to the project and/or suitability should be directed to Dr Mason ([Email Address Removed]). Applications are invited on an on-going basis but early expression of interest is encouraged.
Funding Notes
This project has a Band 3 fee. Details of our different fee bands can be found on our website (https://www.bmh.manchester.ac.uk/study/research/fees/). For information on how to apply for this project, please visit the Faculty of Biology, Medicine and Health Doctoral Academy website (https://www.bmh.manchester.ac.uk/study/research/apply/).
Informal enquiries may be made directly to the primary supervisor.
References
1. Naik S, Zhang N, Gao P, Fisher MT. On the design of broad based screening assays to identify potential pharmacological chaperones of protein misfolding diseases. Current topics in medicinal chemistry. 2012;12(22):2504-22.
2. Zhang T, Baehr W, Fu Y. Chemical chaperone TUDCA preserves cone photoreceptors in a mouse model of Leber congenital amaurosis. Invest Ophthalmol Vis Sci. 2012;53(7):3349-56.
3. Davidson AE, Millar ID, Burgess-Mullan R, Maher GJ, Urquhart JE, Brown PD, Black GC, Manson FD. Functional characterization of bestrophin-1 missense mutations associated with autosomal recessive bestrophinopathy. Invest Ophthalmol Vis Sci. 2011;52(6):3730-6.
4. Shu X, Tulloch B, Lennon A, Vlachantoni D, Zhou X, Hayward C, Wright AF. Disease mechanisms in late-onset retinal macular degeneration associated with mutation in C1QTNF5. Hum Mol Genet. 2006;15(10):1680-9.
5. Liden M, Romert A, Tryggvason K, Persson B, Eriksson U. Biochemical defects in 11-cis-retinol dehydrogenase mutants associated with fundus albipunctatus. J Biol Chem. 2001;276(52):49251-7.
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