Alzheimer’s disease (AD), Parkinson’s disease (PD), and Huntington’s disease (HD)) are characterised by abnormal protein metabolism1, 2 and mitochondrial impairment and dysfunction also accrues with age in these diseases3. We have used an in vitro model that recapitulates the damage to mitochondria that occurs during neurodegenerative illnesses to carry out a high-throughput screen of 7500 genes. We have recently completed a secondary screen and identified thirty druggable genes (can be pharmacologically manipulated) whose role in neurodegenerative disease we wish to investigate further. These genes fall into three broad classes, those that regulate: protein degradation pathways; free radical levels; DNA repair and transcription.
Therapeutic targets will be characterised and their therapeutic potential assessed in collaboration with Takeda UK using human neuronal cells derived from the induced pluripotent stem cells (IPSC) of patients. We already have IPS cells from patients carrying G51D, A53T and AST18 (triplication) mutations synuclein gene. Other human disease models are now available and in addition we have established pharmacological, lentiviral, molecular and biochemical assays5, 6.
There is very strong translational focus, by the end of the PhD genes that can be targeted to mediate neuroprotection would have been characterised. Furthermore, senior scientists at Takeda Cambridge will support the student and it is hoped that pharmacological compounds that can manipulate specific gens/ signalling pathways will be available.
This project offers a unique opportunity for the student to learn molecular (e.g. lentiviral construction, cloning, siRNA/miRNA mediated gene knockdown, qPCR) imaging (confocal, fluorescent microscopy), biochemical (measurements of proteins, RNA and DNA, assays of mitochondrial function) and stem cell biology (culturing of neurons derived from human IPS cells carrying autosomal dominant mutations in genes implicated in Parkinson’s disease). In addition they will collaborate with an industrial partner and gain insight into a drug development program. The student may also gain experience of using genomic analyses7 and will assess the therapeutic and regenerative potential of the compounds in models of Alzheimer’s and Parkinson’s disease.
1. Jucker M, Walker LC. Pathogenic protein seeding in Alzheimer disease and other neurodegenerative disorders. Ann Neurol. 2011 Oct;70(4):532-40. doi: 10.1002/ana.22615. Review.
2. Howarth J.L, Kelly S, Glover C.P.J, Hobson R.J, Chappell, J. Paul. Gallo J.M Cheetham M.E and Uney J.B. Hsp40 molecules that target to the Ubiquitin-Proteasome System decrease inclusion formation in models of Polyglutamine disease Molecular Therapy 2007 Jun;15(6):1100-5 PMID: 17426712
3. Rubinsztein DC, DiFiglia M, Heintz N, Nixon RA, Qin ZH, Ravikumar B, Stefanis L, Tolkovsky A. Autophagy and its possible roles in nervous system diseases, damage and repair. Autophagy. 2005 Apr;1(1):11-22. Epub 2005 Apr 30. Review.
4. Hasson, S et al & Youle, RJ. High-content genome-wide RNAi screens identify regulators of parkin upstream of mitophagy. Nature. 2013Nov 24. doi: 10.1038/nature12748
5. Howarth JL, Lee YB, Uney JB Using viral vectors as gene transfer tools (Cell Biology and Toxicology Special Issue: ETCS-UK 1 day meeting on genetic manipulation of cells). Cell Biol Toxicol. 2010 Feb;26(1):1-20.
6. Scott H, Howarth J, Lee YB, Wong LF, Bantounas I, Phylactou L, Verkade P, Uney JB. MiR-3120 is a mirror microRNA that targets heat shock cognate protein 70 and auxilin messenger RNAs and regulates clathrin vesicle uncoating. J Biol Chem. 2012 Apr 27;287(18):14726-33. Epub 2012 Mar 5.
7. iCLIP identifies novel roles for SAFB1 in regulating RNA processing and neuronal function Rivers CA et al UneyJB. BMC Biology In press.