Human neuron-astrocyte 3D co-culture-based assay for evaluation of neuroprotective compounds

Ana Paula Terrasso, Ana Carina Silva, Augusto Filipe, Pedro Pedroso, Ana Lúcia Ferreira, Paula Marques Alves, Catarina Brito, Journal of Pharmacological and Toxicological Methods, Available online 11 October 2016, ISSN 1056-8719, doi: 10.1016/j.vascn.2016.10.001.

Central nervous system drug development has registered high attrition rates, mainly due to the lack of efficacy of drug candidates, highlighting the low reliability of the models used in early-stage drug development and the need for new in vitro human cell-based models and assays to accurately identify and validate drug candidates.

For the assay validation, we have used Idebenone, an antioxidant benzoquinone, shortchain analogue of coenzyme Q10 ; and positive control for neuroprotection over the tBHP insult. It has been widely investigated for the treatment of Friedreich’s ataxia. The Idebenone treatment led to a reduction in oxidative stress markers (Jaber & Polster, 2015). Consistently with its antioxidant activity, Idebenone prevents lipid peroxidation in isolated brain mitochondria, synaptosomes and human hepatic cells (Erb et al., 2012). It was also shown to protect against ROS-induced damage in several in vitro cultures, as primary cortical neurons and immortalized neural cells. We observed a dose-dependent neuroprotective effect over tBHP insult when Idebenone is administrated in pre-incubation.

The specificity of the neuroprotective effect observed for Idebenone over tBHP insult was evaluated using a non-oxidative lesion induced by chloramphenicol. Idebenone did not confer a neuroprotective effect over the chloramphenicol insult indicating that the neuroprotection observed for tBHP is specific and not an assay artefact.