Parkinson’s disease: A transgenic cell model for studying α-synuclein aggregation and seeding

PLOPER, D; GUAYAN, ML; GONZALEZ LIZARRAGA, MF; TERÁN, MM; TOMAS GRAU, RH; AVILA, CÉSAR L.; SOLIZ SANTANDER, SE; MELLA LOPEZ, R; VILLACÉ LOZANO, P; SALADO, C; CHEHÍN, R

Rosario

Congreso; L Reunión Anual de la Sociedad Argentina de Biofísica; 2022

Sociedad Argentina de Biofísica

Amyloid aggregation of a-synuclein (α-Syn) in dopaminergic neurons is a central hallmarkof Parkinson’s disease (PD). Toxic aggregation of this protein within dopaminergic neuronsis thought to play a key role in the pathogenesis of the disease, and transfer of theseaggregates between cells has been suggested as the primary mechanism of diseasespreading. Here, we present an improved transgenic model for studying α-Synaggregation and seeding in cultured cells. In an SHSY5Y stably overexpressing α-SyntagRFP, we show that treatment with exogenous recombinant human α-Syn preformedfibrils (PFF triggers an increase of the endogenous α-Syn-tagRFP puncta that are positivefor Thioflavin S (ThS), an amyloid specific probe, and phospho-α-Syn (S129), a hallmarkof toxic α-Syn species, suggesting that exogenous fibrils are able to act as seeds for theendogenous protein. In addition, lysosomal biogenesis is also increased. The increase ofthese parameters is specific for α-Syn amyloid fibrils, as treatment with amyloid fibrilsformed with other proteins had no effect on the model. Finally, as proof of concept, weshow how this model can serve to identify novel inhibitors of α-Syn protein aggregation,phosphorylation, seeding and uptake