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Principal Investigator: Tal Somekh
Organization: ERISYON INC
Fiscal Year: 2020
Award: $247,876
Funding agency: National Institute of Neurological Disorders and Stroke
ABSTRACT Parkinson’s disease (PD) is a neurodegenerative disorder that is typically
diagnosed when 60% of a brain region’s dopaminergic neurons have already degenerated. A
strong contender for the elusive molecular biomarker is a neuronal protein - α-synuclein and its
post-translationally modified phosphorylated isoform (pSer-129). Along with other C-terminal
modifications, this modification has been found as the insoluble aggregated and fibrillar form in
the brain tissue of deceased Parkinson’s patients. The progression and disease severity has
been correlated to its relative levels in patient’s cerebrospinal fluids (CSF) as well.
However, current technologies, such as immuno-assays and mass-spectrometry lack the
necessary sensitivity and capability to accurately quantify both the phosphorylated and
unmodified form of the protein simultaneously and in clinically limited samples, such as the
cerebrospinal fluid. This technological limitation has hampered the development of a robust
clinical assay and validation of these implicated neuronal biomarkers.
Fluorosequencing, a new single molecule protein sequencing technology, is a massively
parallelized platform for identifying and quantifying individual proteins in a complex mixture. The
important and distinguishing feature of the single molecule platform is its inherent ability to
absolutely quantify and discriminate the different species of peptides, including the species
differing in the positions and the occupancy of phosphorylated residues.
Through the grant phase I and II, we propose to utilize this platform technology to develop a
clinical assay for measuring the abundances of the different species of α-synuclein from
extremely limited biological samples and correlating the measurements with patient outcomes.
In phase I, we will spike-in modified and unmodified alpha-synuclein species at different
proportions into CSF, establishing assay reproducibility and replicability (aim 1) and obtain limits
of detection and discrimination between the different α-synuclein phosphorylated species (aim
2). Completing these objectives will establish an optimized work-flow and the determination of
the sample requirements necessary for measuring the levels in patient samples. Successful
completion of the two phases would result in a commercialization path towards a laboratory
developed test or diagnostic assay.
Terms: <Address><Age><Amino Acid Sequence Determinations><Animal Model><Animal Models and Related Studies><Assay><Bioassay><Biologic Assays><Biological><Biological Assay><Biological Markers><Brain><Brain Nervous System><Brain region><C-terminal><Cerebrospinal Fluid><Clinical><Cognitive Discrimination><Complex Mixtures><DA Neuron><Degenerative Neurologic Diseases><Degenerative Neurologic Disorders><Detection><Development><Diagnosis><Diagnostic><Diagnostic tests><Discrimination><Disease><Disease Progression><Disorder><Dopamine neuron><Encephalon><Goals><Grant><Human><Immunoassay><Individual><Isoforms><L-Serine><Laboratories><Liquid substance><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Measurement><Measures><Methodology><Modern Man><Modification><NAC precursor><Nerve Cells><Nerve Unit><Nervous System Degenerative Diseases><Neural Cell><Neural Degenerative Diseases><Neural degenerative Disorders><Neurocyte><Neurodegenerative Diseases><Neurodegenerative Disorders><Neurologic Degenerative Conditions><Neurons><PARK1 protein><PARK4 protein><Paralysis Agitans><Parkinson><Parkinson Disease><Parkinson's disease><Parkinsons disease><Pathology><Patient outcome><Patient-Centered Outcomes><Patient-Focused Outcomes><Patients><Peptide Sequence Determination><Peptides><Performance><Phase><Phosphorylation><Position><Positioning Attribute><Post-Translational Modification Protein/Amino Acid Biochemistry><Post-Translational Modifications><Post-Translational Protein Modification><Post-Translational Protein Processing><Posttranslational Modifications><Posttranslational Protein Processing><Primary Parkinsonism><Process><Property><Protein Isoforms><Protein Modification><Protein Phosphorylation><Protein Sequence Determinations><Protein Sequencing><Protein Sequencing Molecular Biology><Proteins><Reagent><Reporting><Reproducibility><SNCA><SNCA protein><Sampling><Serine><Severity of illness><Site><Source><Study models><Technology><Testing><Validation><Variant><Variation><Work><a-syn><a-synuclein><ages><alpha synuclein><alphaSP22><analytical method><bio-markers><biologic marker><biomarker><biomarker development><biomarker discovery><brain tissue><cerebral spinal fluid><clinical applicability><clinical application><clinical biomarkers><clinical relevance><clinically relevant><clinically useful biomarkers><commercialization><degenerative diseases of motor and sensory neurons><degenerative neurological diseases><design><designing><developmental><diagnostic assay><disease severity><dopaminergic neuron><fluid><insoluble aggregate><instrumentation><liquid><model of animal><model organism><molecular biomarker><molecular marker><neurodegenerative illness><neuronal><non A-beta component of AD amyloid><non A4 component of amyloid precursor><novel><protein aggregate><protein aggregation><protein biomarkers><protein complex><protein markers><single molecule><spinal fluid><synucleinopathy><α-syn><α-synuclein>