P3.13. INHIBITION OF ALPHA-SYNUCLEIN AGGREGATION AND ER STRESS RESCUES NEURODEGENERATION IN A 3D IN VITRO MODEL OF PARKINSON’S DISEASE
Natalia Siwecka1,2, Michał Golberg3,4, Grzegorz Galita1, Wioletta Rozpędek-Kamińska1, Ireneusz Majsterek1
1 Medical University of Lodz, Department of Clinical Chemistry and Biochemistry, 5 Mazowiecka St., 92-215 Lodz, Poland
2 Medical University of Lodz, Clinical Department of Neurology with Stroke Treatment Unit, University Clinical Hospital No. 1, 22 Kopcinskiego St., 90-153 Lodz, Poland
3 Medical University of Lodz, Department of Histology and Embryology, 7/9 Zeligowskiego St., 90-752 Lodz, Poland
4 Medical University of Warsaw, Department of Child Psychiatry, Children's Clinical Hospital, 63A Zwirki i Wigury St., 02-091 Warsaw, Poland
INTRODUCTION: Parkinson’s disease (PD) is the second most common neurodegenerative disorder. The major molecular event underlying the pathophysiology of PD is the abnormal accumulation of α-synuclein (α-syn) within the dopaminergic neurons of the midbrain, which results in the induction of endoplasmic reticulum (ER) stress conditions and PERK-mediated neural cell apoptosis.
AIM(S): The main objective of the present study was to determine the potential effect of the small-molecule inhibitors of α-syn aggregation and ER stress-mediated PERK signaling pathway against neurodegeneration in a novel, 3D in vitro model of PD.
METHOD(S): The effectiveness of the selected α-syn aggregation inhibitor (anle138b) and PERK inhibitor (AMG44) was assessed in a midbrain organoid model of PD derived from a human iPSC line. Neurodegeneration was induced via incubation with the neurotoxin 6-hydroxydopamine (6-OHDA) and α-syn pre-formed fibrils (PFF). Cell viability was assessed by the CellTiter-Glo 3D assay, and the presence of α-syn aggregates was detected by immunofluorescence.
RESULTS: The cell viability analysis demonstrated a significant increase in ATP production in PD organoids treated with anle138b and AMG44, with the most prominent effect upon combining the two inhibitors. Immunofluorescence analysis revealed a significant decrease in phospho-α-syn and aggregated-α-syn levels after treatment with the compounds, and combining anle138b with AMG44 resulted in the greatest anti-aggregative effect.
CONCLUSIONS: Combination therapy with the selected α-syn and ER stress inhibitors effectively rescues neurodegeneration in the 3D in vitro model of PD. The results obtained may help develop the first disease-modifying therapy for PD.
FINANCIAL SUPPORT: This work was supported by the PRELUDIUM BIS 3 grant no. 2021/43/O/NZ5/02068 from the National Science Centre, Poland, and by the grant of the Medical University of Lodz, Poland, no. 503/1-156-07/503-11-001.