Evaluating the use of microelectrode array technology and cell-based neuronal culture models for proconvulsant risk assessment: progress from the HESI NeuTox consortium

Millard, Daniel C.; Nicolini, Anthony M.; Arrowood, Colin; Hayes, Heather B.; Bradley, Jenifer A.; Strock, Christopher J.; Freudenrich, Theresa M.; Wallace, Kathleen; Shafer, Timothy J.; Cato, Matthew J.; Chaudhary, Khuram W.; Saavedra, Lorena; Haag, Daniel; Feaster, Tromondae K.; Anson, Blake D.; Luerman, Greg; Hess, Dietmar; Shi, Yunfeng; Li, Jinqing; Shi, Hong; Lévesque, Paul; Kreir, Mohamed; Lü, Hua; Suzuki, I.; Odawara, Aoi; Miyamoto, Naokazu; Ojima, Atsuko; Kanda, Yasunari; Blinova, Ksenia; Herr, David W.; Roberts, R.C.; Pierson, Jennifer

doi:10.1016/j.vascn.2019.05.076

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“…Current attempts to develop improved neurotoxicity assays tackle these two issues in different ways (Schultz et al 2015). The main options for test systems are rat primary neurons (Forsby et al 2009;Sandström et al 2017;Bradley et al 2018;Kreir et al 2018;Millard et al 2019), cells differentiated from pluripotent stem cells (Pei et al 2016;Sherman and Bang 2018;Sirenko et al 2019;Tukker et al 2020;Brüll et al 2020), and cell lines (Forsby et al 2009;Krug et al 2013;Stiegler et al 2011;Klima et al 2020). The latter often have the disadvantage that they do not form effective synapses.…”

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confidence: 99%

Human neuronal signaling and communication assays to assess functional neurotoxicity

et al. 2020

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Prediction of drug toxicity on the human nervous system still relies mainly on animal experiments. Here, we developed an alternative system allowing assessment of complex signaling in both individual human neurons and on the network level. The LUHMES cultures used for our approach can be cultured in 384-well plates with high reproducibility. We established here high-throughput quantification of free intracellular Ca2+ concentrations [Ca2+]i as broadly applicable surrogate of neuronal activity and verified the main processes by patch clamp recordings. Initially, we characterized the expression pattern of many neuronal signaling components and selected the purinergic receptors to demonstrate the applicability of the [Ca2+]i signals for quantitative characterization of agonist and antagonist responses on classical ionotropic neurotransmitter receptors. This included receptor sub-typing and the characterization of the anti-parasitic drug suramin as modulator of the cellular response to ATP. To exemplify potential studies on ion channels, we characterized voltage-gated sodium channels and their inhibition by tetrodotoxin, saxitoxin and lidocaine, as well as their opening by the plant alkaloid veratridine and the food-relevant marine biotoxin ciguatoxin. Even broader applicability of [Ca2+]i quantification as an end point was demonstrated by measurements of dopamine transporter activity based on the membrane potential-changing activity of this neurotransmitter carrier. The substrates dopamine or amphetamine triggered [Ca2+]i oscillations that were synchronized over the entire culture dish. We identified compounds that modified these oscillations by interfering with various ion channels. Thus, this new test system allows multiple types of neuronal signaling, within and between cells, to be assessed, quantified and characterized for their potential disturbance.

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