1. Design and characterisation of peptides for diagnostic and therapeutic applications in Parkinson's disease
- Author
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Watt, Kathryn, Mason, Jody, Williams, Robert, and James, Tony
- Abstract
Parkinson's Disease (PD) is the second most common neurodegenerative disease, accounting for approximately 15% of all dementias. However, despite its prevalence, there is currently no disease-modifying or neuroprotective treatments available. All of the currently available treatments aim to ease the symptoms associated with the disease, but do not slow or change the disease pathway. There are several reasons for the lack of disease-modifying treatments, and these include the incomplete understanding of the disease; the current incapacity to identify individuals with an early form of the disease when drug intervention may be more impactful, and the inability to effectively diagnose or monitor disease progression at a biological level. a-synuclein (aS) is a key protein in PD pathology, with several single-point mutations having been identified in familial cases of PD. Herein, aS aggregation properties are studied in the presence of DMPS lipid vesicles for wild-type aS and five of the most predominant single point missense mutants associated with early-onset PD. Experiments presented here highlight significant differences between aggregation rates, the number of aggregates produced, and overall fibril morphologies of aS in the presence of lipid vesicles. These findings have important implications regarding the interplay between the lipids required for aS function and the individual point mutations known to accelerate PD and related diseases. Following this, we build on previous work which identified the peptide 4554W via a semi-rational intracellular peptide-library screen, by expanding this screen to identify peptides that target five of the most prominent early-onset familial variants. All of the identified peptides reduced aggregation of the respective target, with several also identified to be effective at reducing aggregation when incubated with other variants. In addition, our results demonstrate that the previously optimised peptide, 4554W(N6A), is highly effective against not only WT aS, but also several of the single-point mutant forms, and hence is a suitable baseline for further work towards a PD therapeutic. Finally, to explore the possibility of developing a method of early diagnosis, or tools to monitor disease progression, several fluorophore-labelled peptides were designed, synthesised, and evaluated for their potential to act as chemical tools that can assist in the identification of different oligomeric species. Labelling the 4554W peptide at position K1 with an environmentally-sensitive fluorophore was found to result in the most effective probe across all experiments. This probe was found to be more sensitive to pre-fibrillar species in the aS aggregation pathway than the widely used fluorescent probe ThT. Moreover, the probe was found to produce a differential response in the presence of various aS oligomer samples, indicating that it may be useful as an aS-oligomer probe.
- Published
- 2022