Objective: This research line aims to perform a compound screening on the prophylactic capacity of a high number of nutraceuticals and drugs to prevent dopaminergic neuron (DOPAn) demise in our new slow progressive in vitro degenerative model of Parkinson’s Disease (PD).

Background: PD is the second most common neurodegenerative disease in the world and is expected to double in 2040 due to population aging.

Currently, diagnosis depends on the presence of motor symptoms, which manifest when 50% or more of DOPAns in the substantia nigra are dead, even if it is recognized that the degeneration process starts decades before the onset of symptoms.

Inflammation is gaining attention as a PD trigger. From our previous data we identified TNF-α as the determinant of DOPAn demise in the pre-diagnostic phase of the disease.

To address the most effective way to prevent progression of the disease need a solid, reproducible, easy-to-handle and rapid in vitro model of slow-dying DOPAn, to mimic earlier stages of PD.

Method: This was done by use of:

-SH-SY5Y (SH) human cell line was differentiated into DOPAn-SH by adding arachidonic acid (RA) and phorbol 12-myristate 13-acetate (PMA) [1].

-The slow degeneration in DOPAn-SH was obtained by low doses of rotenone (rot).

-Anti–inflammatory compounds to test were selected through literature research.

-Treatments were organized based on a priority approach.

Results: SH cells were successfully differentiated into DOPAn (immunofluorescence, qRT-PCR validation) and addition of rot induced a time dependent reduction of DOPAn-SH viability (70% at 2 days, p<0.001; 30% at 6 days, p<0.001). Among the 50 compounds selected, the anti-TNF-α antibody infliximab prevents DOPAn-SH cell demise (% cell viability: 101%, not statistically significant vs non treated cells; p<=0.001 vs rot treated cells), confirming the leading role of TNF-α as damage trigger. Curcumin showed a dose-dependent protection with higher efficacy at 5 µM concentration (cell viability: 39% at 20 µM, 53% at 10 µM, 75% at 5 µM vs 52% in rot treated cells), a result capable of improvement.

Conclusion: In conclusion DOPAn-SH is a powerful tool for drug repurposing and nutraceuticals screening to use before moving to the more complex systems available in the laboratory. Therefore, we confirmed the detrimental role of TNF-α and the potential of nutraceuticals in fighting it.

References: [1] Barth, M., Toto Nienguesso, A., Navarrete Santos, A. et al. Quantitative proteomics and in-cell cross-linking reveal cellular reorganisation during early neuronal differentiation of SH-SY5Y cells. Commun Biol 5, 551 (2022). https://doi.org/10.1038/s42003-022-03478-7.

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MDS Abstracts - https://www.mdsabstracts.org/abstract/the-challenge-to-prevent-parkinsons-disease-with-a-new-in-vitro-approach/