Authors

P Villace¹; AC Salado¹; RM Mella¹; J Gamiz¹; M Roura¹; 
¹ Innoprot, Spain

Discussion

Amyotrophic Lateral Sclerosis disease (ALS) is characterized by the death of both upper and lower motor neurons in the motor cortex of the brain, the brain stem, and the spinal cord. Prior to their destruction, motor neurons develop intracellular protein inclusions in their cell bodies and axons. These inclusions often contain ubiquitin, and generally incorporate one of the ALS-associated proteins: SOD1, TAR DNA binding protein (TDP-43, or TARDBP), or FUS. Innoprot has developed a novel fluorescence cell-based assay in U2OS cells that expresses constitutively the FUS protein in a TPD43-induced model. This cellular model allows the study of the relationship between FUS and TDP-43 proteins aggregation pathways in the development of the disease. This novel ALS fluorescence cell-based assay has been designed for High Content Screening applications to find compounds able to inhibit or modulate TDP43/FUS aggregation after severe cytotoxic damage induction by Arsenite. In this work, this model was used to screen a library of 880 compounds. ISRIB and LiCl compounds were used as positive protection controls in the fluorescent TDP43/FUS aggregation model. After the screening campaign, positive compounds were chosen for further testing, based on the strength of the initial response and the lack of cytotoxicity. Our results indicated that the pharmacological inhibition or modulation of TDP43 and FUS aggregation implicated in ALS is a valid strategy for drug screening.