Multimarker Parkinson's Diagnotic test kit for monitoring disease progression based on electroanalytical detection of protein changes in blood.

In England alone, dementia affects more than 600,000 people with Parkinson's pathology constituting a significant and growing part of this. These numbers are increasing as the mean population age grows and represent a very substantial burden on the welfare system, the NHS and families. Parkinson's disease is a heterogeneous disease characterised by progressive neuronal loss, causing a decline in movement and other functions. Up to 80% of long-term survivors with Parkinson's develop dementia whereas a significant percentage reach early motor disability such as postural instability and falls. Currently there is no disease-modifying therapy, which in part is hindered by the lack of an objective marker that stratifies these patient subgroups and objectively measures their disease progression. Such a biomarker is urgently needed to facilitate the development of clinical trials that aim to slow down neurodegeneration and importantly to also inform prognosis and better plan care. Based on our preliminary work in serum samples, our prediction is that such a biomarker can be developed with technology that enables multiplexed sampling and quantitation of several disease-specific reactive protein changes that are present in low abundance in the peripheral circulation. This approach requires the identification of innovative biomarker-candidates and the availability of ultrasensitive, label-free protein detection methods. The current methods of screening proteins involve expensive and highly specialized pieces of equipment or are prone to low levels of sensitivity and/or complicated analytical procedures associated with significant (up to 200%) analytical error. Electrical detection methodologies are portable, highly sensitive, cheap, high throughput (measurement time of minutes - particularly important if many samples are being screened) and multiplexable (multiple proteins detected simultaneously giving, in relevant cases, a "fingerprint" of health). The interfacing of man-made electronics with biological receptor molecules can enable the specific and calibrated detection of markers of disease. Devices built around these principles have already had a profound impact on clinical diagnostics and the quality of life of those unfortunate enough to live with chronic diseases such as diabetes. An assessment of protein levels in biological fluid (urine, saliva, blood serum, spinal fluid) constitutes a critical reflection of current health and may be reflective of the disease progression. We have already shown that one of the body's responses to the small protein (alpha-synuclein) that accumulates in the brains of patient with Parkinson's disease is to generate anti-alpha-synuclein antibodies, which we have measured and correlated with the disease stage using electrochemistry. We have also shown that circulating microvesicles in the serum of patients have distinct bioactivity and protein composition. We propose to measure these microvesicle-associated proteins by integrated microfluidic multiplexed devises and in combination with our earlier data on auto-antibodies develop a multi-parameter kit to monitor disease progression. We are uniquely positioned to develop this technology and provide proof of concept in two extensively characterized longitudinal patient cohorts. Specifically, we will ask whether a combination of protein markers reflect the rate of progression to dementia or severe movement disability in carefully selected patient samples by correlating serial serum levels with detailed clinical assessments. In summary, we are seeking to solve a profound clinical challenge in the area of neurodegeneration by developing a unique multi-parameter receptor chemistry device that we believe will have unprecedented application and potency.

Parkinson's disease (PD) is the second most common neurodegenerative disease with increasing prevalence. It is clinically heterogeneous disease with variable rates of progression to dementia or motor disability. Currently there is no cure or objective markers of disease progression. Thus, there is an urgent unmet need for a biomarker that can be easily used in the clinic to stratify patients for clinical trials of disease modifying therapies and inform prognosis. Our prediction is that such a biomarker can be developed with technology that enables multiplexed sampling and quantitation of several disease-specific reactive protein changes that are present in low abundance in the peripheral circulation. The "standard" approach to quantifying proteins in patient samples has been ELISA. This is a multistep, laborious analysis that, though very sensitive, can be associated with significant error (two antibodies must be utilized per target, one of which must be associated with a reproducibly amplifying enzyme) and background noise. When coupled to the native (non-disease related) scatter in background marker levels amongst a real population an ability to reliably differentiate between disease and control becomes more difficult still. Instead, we focus on multiplexed strategies in which several markers are assayed sensitively and simultaneously. In preliminary work, we have been able to show the utility of electrochemistry in tracking PD disease status by measuring autoantibodies against alpha-synuclein. We have also identified proteins in circulating microvesicles that are specifically enriched in PD. We have developed, and continue to develop, entirely new impedance and capacitance analytical methods, which are readily integrated into designed microfabricated chips. We have also been able to demonstrate that surface chemical methods can be employed in facilitating the reliable, and entirely unlabeled, single step detection of markers in patient samples. By combining expertise in Neurology and Chemistry, this proposal seeks to extend these innovative developments to (1) integrate new capacitative and sample pre-treatment methods with the simultaneous accurate quantification of multiple promising serum markers and (2) provide proof of concept using two patient cohorts, that a multi-parameter kit can provide a simple and cost-effective biomarker to monitor disease progression in PD.
The work will be published in high impact peer-reviewed journals and discussed at conferences in the UK and overseas. The project team members have a strong track record in the effective execution of multi-disciplinary projects and it will be under this umbrella that employed postdocs will work. We would fully expect that they would find the project demanding, highly stimulating, rewarding and productive. The analytical, planning, communication and presentational skills of these young researchers, evident at periodic group and cross-group meetings, will be polished through the duration of the project. The PI and the Co-I have experience in the commercialisation of laboratory research, and are, accordingly, familiar with issues of scale up, cost and intellectual property. Several aspects of the project benefit from underlying patent protection. It is envisaged that IP will be generated through the project. In previous months a number of large Diagnostic/Pharma companies have expressed considerable interest in precursors to the project. We will continue to be engaged in discussions with the commercial sector throughout its duration.