A novel cellular model to screen for inhibitors α-Synuclein aggregation and phosphorylation

GONZÁLEZ LIZARRAGA, MARÍA FLORENCIA; PLOPER, DIEGO; SOCIAS, SERGIO BENJAMÍN; TOMAS-GRAU, RODRIGO; SEQUEIRA, SABRINA; MICHEL, PATRICK P.; MELLA LÓPEZ, ROSA MARÍA; VILLACÉ LOZANO, PATRICIA; SALADO, CLARISA; RAISMAN-VOZARI, RITA; CHEHÍN, ROSANA NIEVES; ÁVILA, CÉSAR LUIS

Rosario

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

Sociedad Argentina de Biofísica

Synucleinophaties are characterized by the abnormal accumulation of alpha-synuclein (αSyn) aggregates in vulnerable neuron and glia cells. The spread of these aggregates throughout the nervous system correlates with the progression of the disease, and thus they constitute a promising target to develop disease‐modifying therapies. The growth and maturation of these aggregates within the cell involves a complex interplay between uptake, fibrillization, post-translational modifications, and interactions with membranous organelles. Herein we present a cellular model that recapitulates many pathological events associated with αSyn accumulation. Our model relies on the treatment of transgenic SH-SY5Y cells overexpressing an αSyn-tRFP fusion protein (INNOPROT, Derio, Spain) with preformed fibrils of αSyn. The fluorescence tag within intracellular αSyn allows to easily monitor aggregation in living cells using fluorescence microscopy. We demonstrate that the intracellular aggregates have an amyloid like nature as revealed by ThS staining. The aggregates also show pathological phosphorylation on Ser129 whichis the predominant modification of alpha-synuclein in Lewy bodies. We propose that this model could be used as a platform to screen candidate drugs for therapeutic intervention in PD and other synucleinopathies.