A microscopy-based small molecule screen in primary neurons reveals neuroprotective properties of the FDA-approved anti-viral drug Elvitegravir.
Glutamate toxicity is a pathomechanism that contributes to neuronal cell death in a wide range of acute and chronic neurodegenerative and neuroinflammatory diseases. Activation of the N-methyl-D-aspartate (NMDA)-type glutamate receptor and breakdown of the mitochondrial membrane potential are key events during glutamate toxicity. Due to its manifold functions in nervous system physiology, however, the NMDA receptor is not well suited as a drug target. To identify novel compounds that act downstream of toxic NMDA receptor signaling and can protect mitochondria from glutamate toxicity, we developed a cell viability screening assay in primary mouse cortical neurons. In a proof-of-principle screen we tested 146 natural products and 424 FDA-approved drugs for their ability to protect neurons against NMDA-induced cell death. We confirmed several known neuroprotective drugs that include Dutasteride, Enalapril, Finasteride, Haloperidol, and Oxybutynin, and we identified neuroprotective properties of Elvitegravir. Using live imaging of tetramethylrhodamine ethyl ester-labelled primary cortical neurons, we found that Elvitegravir, Dutasteride, and Oxybutynin attenuated the NMDA-induced breakdown of the mitochondrial membrane potential. Patch clamp electrophysiological recordings in NMDA receptor-expressing HEK293 cell lines and primary mouse hippocampal neurons revealed that Elvitegravir does not act at the NMDA receptor and does not affect the function of glutamatergic synapses. In summary, we have developed a cost-effective and easy-to-implement screening assay in primary neurons and identified Elvitegravir as a neuro- and mitoprotective drug that acts downstream of the NMDA receptor.
['Merz|Simon F|SF|0000-0002-8056-0766', 'Bengtson|C Peter|CP|0000-0002-8245-3323', 'Tepohl|Clara|C|', 'Hagenston|Anna M|AM|0000-0001-7747-5336', 'Bading|Hilmar|H|0000-0003-0258-4348', 'Bas-Orth|Carlos|C|0000-0002-6037-8350']
D000818:Animals D000998:Antiviral Agents D016923:Cell Death D002478:Cells, Cultured D000075402:Channelrhodopsins D017277:Drug Approval D019706:Excitatory Postsynaptic Potentials D057809:HEK293 Cells D006801:Humans D053078:Membrane Potential, Mitochondrial D008810:Mice, Inbred C57BL D008853:Microscopy D016202:N-Methylaspartate D009474:Neurons D000066829:Neuroprotection D018696:Neuroprotective Agents D062308:Optogenetics D015363:Quinolones D018091:Receptors, AMPA D017470:Receptors, Glutamate D016194:Receptors, N-Methyl-D-Aspartate D054852:Small Molecule Libraries D013569:Synapses D014481:United States D014486:United States Food and Drug Administration
Neuroprotection Glutamate toxicity NMDA receptor Mitochondria Dutasteride Finasteride Oxybutynin Channelrhodopsin Excitotoxicity