Study Rationale:
The amplification of small quantities of misfolded protein aggregates from clinical samples offers an attractive route towards developing sensitive diagnostics for a range of protein misfolding diseases, including Parkinson’s. The development of seed amplification assays (SAAs) targeting alpha-synuclein in Parkinson’s disease represents a major advancement, yet, current alpha-synuclein SAAs often provide only binary (positive/negative) outcomes, lacking the resolution required to monitor disease progression or evaluate the effectiveness of therapeutic interventions. To address this challenge, we have recently leveraged the power of microfluidics to develop a digital seed amplification assay (dSAA), allowing for direct quantification of seed burden in complex biological solutions.
Hypothesis:
In the current implementation, quantification of CSF seed numbers is readily available using the dSAA platform, however, a direct correlation between seeds found in in different brain regions and their correlation with PD progression is still lacking.
Study Design:
We will employ dSAA optimisation steps, including different solution conditions (substrate, pH, ionic strength) to provide a fully reconstructed image of aSyn CSF seed load, CNS soluble aggregates and finally LB/LN load and Braak staging by further optimisation of the dSAA protocols will be implemented to provide high accuracy discrimination of dSAA data at varying disease progression stages.
Impact on Diagnosis/Treatment of Parkinson’s disease:
Through this biomarker discovery approach we will be able to provide a novel method for fingerprinting the correlation between CSF and brain seeds as a means to not only determine PD vs healthy patients, but go beyond the state of the art in providing access to early diagnosis of PD.
Next Steps for Development:
With the support of this grant, we will build a world class team and allow the generation of custom-made optical platform which will be dedicated to moving forwards this ambitious, challenging but exciting technology development project on short timescales towards a true dSAA platform for practical use.