Targeting the earliest steps in the aggregation process of the intrinsically unfolded protein alpha-Synuclein, the most validated target in Parkinson’s disease.

α-Synuclein is a small presynaptic protein whose aggregation and fibrillation are considered hallmarks of Parkinson’s disease. Many studies have focused on the fibrillation process of α-synuclein, however early phase small oligomers are considered the main toxic species. The spread of disease is believed to happen by the transmission of small oligomers or pre-fibrils between cells to infect healthy neurons.
In the Auer lab a novel, unique micro-bead based confocal fluorescence assay has been developed, named ASYN-CONA, to investigate the earliest oligomerization steps of α-synuclein. The assay relies on two differently labelled α-synucleins, a green one attached to microbeads and the red α-synuclein incubated with the beads in solution. Aggregation is induced on bead and followed by automated and quantitative detection of the red fluorescence emission imaged as “halos” or “rings” formed around the microbeads by confocal scanning on the Perkin Elmer Opera instrument. As this bead-based assay measures the initial linear aggregation phase of α-synuclein, rather than the formation of later stage fibrils, monitored in other assays, it opens new opportunities for the identification of early stage aggregation inhibitors.
The PhD project will involve various translational and basic science steps.
Translational and drug discovery projects aspects: (a) Using the proprietary small molecule, fragment and phage libraries of the Auer lab, new binders and inhibitors will be identified. (b) Any hit compounds will be further explored in induced pluripotent stem cell (iPSC) models of the Kunath lab (Devine et al. 2011), included CRISPR-engineered of Parkinson’s (Singh Dolt et al., 2017).
Basic Science projects aspects: (c) The protein biochemical and structural characterisation as well as the evaluation of fibril forming potential of the oligomers formed on bead is required for a proper understanding of the neurological relevance of on-bead aggregates and the inhibitors found in the assay. A combination of biophysical techniques, mainly single molecule spectroscopy and biochemical and cellular imaging will be applied to produce mechanistic and biological evidence.
This project will be performed in close collaboration between Kunath and Auer labs who have worked together for over 5 years, including co-supervision of a PhD student. This will give the student an excellent opportunity to cover all aspects of an early drug discovery project, including chemical synthesis and analysis, fluorescence labelling, protein expression, purification and mutation, confocal fluorescence spectroscopy and imaging, as well as experience in testing hit compounds in human neuronal models in the Kunath lab. In addition to multiple opportunities for high quality publications, the student will acquire interdisciplinary knowledge in one of sciences most active fields of targeting intrinsically unfolded proteins. Career progression in academia and industry will be possible after completing the PhD.

Further Information about various assay and screening platforms developed and run by the Auer lab can be found on: https://sites.google.com/view/the-auer-lab-uoe/home.
The manuscript about ASYN-CONA assay is currently under review. The related UPS-CONA assay technique provides a good overview of technology as well as hard and software used (BMC Biology (2018) 16:88 https://doi.org/10.1186/s12915-018-0554-z).
Kunath Lab: http://www.crm.ed.ac.uk/research/group/mechanisms-neurodegeneration.