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

GONZALEZ LIZARRAGA, MF; TOMAS GRAU, RH; MELLA LOPEZ, R; RAISMAN VOZARI, R; SOCIAS, B; PLOPER, D; MICHEL, PATRICK P.; SEQUEIRA, S; SALADO, C; VILLACÉ LOZANO, P; AVILA, CÉSAR L.; CHEHÍN, R

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 aggregatesthroughout the nervous system correlates with the progression of the disease, and thusthey constitute a promising target to develop disease‑modifying therapies. The growthand maturation of these aggregates within the cell involves a complex interplay betweenuptake, fibrillization, post-translational modifications, and interactions with membranousorganelles. Herein we present a cellular model that recapitulates many pathologicalevents associated with αSyn accumulation. Our model relies on the treatment oftransgenic SH-SY5Y cells overexpressing an αSyn-tRFP fusion protein (INNOPROT, Derio,Spain) with preformed fibrils of αSyn. The fluorescence tag within intracellular αSynallows to easily monitor aggregation in living cells using fluorescence microscopy. Wedemonstrate that the intracellular aggregates have an amyloid like nature as revealed byThS staining. The aggregates also show pathological phosphorylation on Ser129 whichis the predominant modification of alpha-synuclein in Lewy bodies. We propose that thismodel could be used as a platform to screen candidate drugs for therapeutic interventionin PD and other synucleinopathies.