As shown previously by AstraZeneca in Nature reviews*, one third of the safety failures is linked to central nervous system (CNS) toxicity observed during the clinical trials.

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* Cook D, Brown D, Alexander R, March R, Morgan P, Satterthwaite G, Pangalos MN (2014) Lessons learned from the fate of AstraZeneca’s drug pipeline: a five-dimensional framework. Nat Rev Drug Discov. 13(6):419-31.

To avoid this attrition, the potential neurotoxicity of new chemical entities should be checked as soon as possible during the drug discovery process

To learn more on these assays, please refer to our posters that have been presented in different conferences:

Neurotoxicity assay on neuronal cultures

Study of compounds’ toxicity on neurons

This assay can be performed by cell imaging in HCA/HCS on brain cells, including neuronal cell lines like SH-SY5Y and Luhmes, or iPSC-derived human brain cells . Neurotoxicity assays are performed by treating neuronal cells during at least 48 hours with the tested compounds.

Requirements

  • Compounds: powder or stock solution
  • Cells:  differentiated SH-SY5Y and Luhmes human cell lines, human iPSC-derived cells (PERI4U® as peripheral neurons, CNS4U® as a mix of neurons and astrocytes), or other neuronal cultures
  • Plate format: 96 or 384-well plates

Example of colchicine

Differentiated SH-SY5Y and CNS4U® cells (iPSC-derived mix of human brain cells) were exposed to increasing concentrations of colchicin during 48 hours in 384-well plates. After the treatment, cells were fixed and stained for ß3-tubulin (for neurite length measurement) and Hoechst (for nuclei vizualisation) for fluorescence microscopy observation. Images were analyzed with MetaXpress software (Molecular Devices) to assess the cell numbers and neurite lengths in all cultures.

Impaired communication between neurons can occur by the loss of neurites at lower doses than neuronal cell death! This assay can be multiplexed for the assessment of other parameters in parallel (mitochondrial staining, etc.)

Don’t hesitate to ask for our short report on neurotoxicity assessment using SH-SY5Y and human iPSC-derived brain cells exposed to our in-house neurotoxicity library!