Clinical diagnosis of Parkinson's disease (PD), multiple system atrophy (MSA) and dementia with Lewy bodies (DLB) is challenging, especially in the early stages. Each disease is associated with distinct conformers of misfolded alpha-synuclein (maS) which form typical protein aggregates in the brain and represent key disease biomarkers. Thus, detection and characterization of intracerebral maS aggregates allow a definite diagnosis. The recent development of ultrasensitive assays enabled the detection of maS and other potential new biomarkers in peripheral tissues, although with several limitations. Here, the investigators propose to combine the expertise of leading and young researchers in the field of neurology, structural and molecular biology, biophysics and machine learning to perform ultrasensitive and multi-omics analyses of olfactory mucosa (OM), blood and urine of PD, MSA and DLB patients for detecting and characterizing key peripheral biomarkers allowing accurate disease recognition.

OM, blood and urine likely contain key specific biomarkers of PD, MSA and DLB detectable with specialized ultrasensitive techniques, perhaps in the early disease stages. Therefore, by combining for the first time the RT-QuIC, Simoa SR-X, ELISA and NTA analyses of OM, blood and urine and by exploiting TEM and solution/solid-state NMR analyses, the investigators will attempt at identifying specific fingerprints of alpha-synucleinopathies usefult to stratify living patients. Supported by statistical analyses and customized machine learning algorithms that will combine clinical (including the analysis of olfactory functions) and experimental data, the investigators project could significantly impact the clinical diagnosis of alpha-synucleinopathies.

The investigators project will combine innovative and ultrasensitive diagnostic approaches to evaluate whether OM, blood and urine might serve as non-invasive and easy-to-get samples for biomarkers detection, allowing recognition and stratification of alpha-synucleinopathies. Multidisciplinary collaboration between researchers with specific and complementary skills is required to achieve the goals of the project and to have an impact on future scientific use, clinical applications and wellbeing of patients suffering from these diseases. The investigators project will combine cutting-edge technologies to improve the power of multidimensional biomarker patterns for the clinical diagnosis of PD, MSA and DLB by using easily and periodically collectible tissues. In particular, the investigators will determine unique insights into several key aspects of alpha-synucleinopathies by assessing whether: (1) the OM, blood and urine of PD, MSA and DLB patients contain peripheral maS exploitable to improve the patiens clinical diagnosis/recognition; (2) the peripheral distribution of maS in OM, blood and urine differs between diseases or even among tissues collected from the same patient; (3) the biochemical, morphological and structural features of the RT-QuIC generated aggregates enable discrimination of these diseases; (4) the levels of some key proteins (e.g. NfL, alpha-synuclein, synapsin-3) or structures (e.g. EVs), known to be altered in the blood, are also modified in the OM or urine and allow disease recognition, especially when analyzed in combination with the other ultrasensitive assays; (5) the olfactory impairment can be exploited as indicator of evolving alpha-synucleinopathy and whether it correlates with maS accumulation in OM. The investigators research is transformative and rapidly translatable into clinical practice. The possibility to accurately identify PD, MSA and DLB will bring important advancements in patient selection for emerging pharmacological treatments and clinical trials (better care for people). In addition, the possibility to periodically collect OM, blood and urine samples could consent to monitor disease progression and to evaluate the therapeutic effects in patients under pharmacological treatments. The Investigators findings could also contribute to further investigating whether PD, MSA and DLB are caused by distinct conformers of maS and verify if these latter possess differential tropism for peripheral tissues, eventually unveiling different molecular pathways involved in their pathogenesis. Finally, the investigators approach could have a major impact on counseling and prevention strategies for patients in the prodromal stages of alpha-synucleinopathies, representing a key goal on the path to disease-modifying and neuroprotective therapies before full-blown phenotypes have manifested.

Inclusion Criteria:

• PD (Postuma et al., 2015), MSA (Palma et al., 2018), DLB (McKeith et al., 2017), OND including acquired and hereditary ataxic syndrome with known etiology, progressive supranuclear palsy, corticobasal degeneration, and motor neuron disease.

Exclusion Criteria:

• na

PD (n=20), MSA (n=20), DLB (n=20), OND (n=10, including acquired and hereditary ataxic syndrome with known etiology, progressive supranuclear palsy, corticobasal degeneration, and motor neuron disease) and HS (n=10).

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