Global Safety Pharmacology, Predictive & Investigative Translational Toxicology, Nonclinical Safety, Janssen Research and Development, A Division of Janssen Pharmaceutica NV, Beerse, Belgium.
Global Safety Pharmacology, Predictive & Investigative Translational Toxicology, Nonclinical Safety, Janssen Research and Development, A Division of Janssen Pharmaceutica NV, Beerse, Belgium.
Eur J Pharmacol. 2022 Sep 15;931:175189. doi: 10.1016/j.ejphar.2022.175189. Epub 2022 Aug 18.
Functional network activity is a characteristic for neuronal cells, and the complexity of the network activity represents the necessary substrate to support complex brain functions. Drugs that drastically increase the neuronal network activity may have a potential higher risk for seizures in human. Although there has been some recent considerable progress made using cultures from different types of human-induced pluripotent stem cell (hiPSC) derived neurons, one of the primary limitations is the lack of - or very low - network activity.
In the present study, we investigated whether the limited neuronal network activity in commercial hiPSC-neurons (CNS.4U®) is capable of detecting drug-induced potential seizure risks. Therefore, we compared the hiPSC-results to those in rat primary neurons with known high neuronal network activity in vitro.
Gene expression and electrical activity from in vitro developing neuronal networks were assessed at multiple time-points. Transcriptomes of 7, 28, and 50 days in vitro were analyzed and compared to those from human brain tissues. Data from measurements of electrical activity using multielectrode arrays (MEAs) indicate that neuronal networks matured gradually over time, albeit in hiPSC this developed slower than rat primary cultures. The response of neuronal networks to neuronal active reference drugs modulating glutamatergic, acetylcholinergic and GABAergic pathways could be detected in both hiPSC-neurons and rat primary neurons. However, in comparison, GABAergic responses were limited in hiPSC-neurons.
Overall, despite a slower network development and lower network activity, CNS.4U® hiPSC-neurons can be used to detect drug induced changes in neuronal network activity, as shown by well-known seizurogenic drugs (affecting e.g., the Glycine receptor and Na channel). However, lower sensitivity to GABA antagonists has been observed.
功能网络活动是神经元细胞的特征，网络活动的复杂性代表了支持复杂脑功能的必要基础。大幅增加神经元网络活动的药物可能会对人类癫痫发作的风险更高。尽管最近使用来自不同类型的人诱导多能干细胞（hiPSC）衍生神经元的培养物取得了一些相当大的进展，但主要限制之一是缺乏或非常低的网络活动。
在本研究中，我们研究了商业 hiPSC 神经元（CNS.4U®）中的有限网络活动是否能够检测到药物引起的潜在癫痫发作风险。因此，我们将 hiPSC 结果与具有已知高神经元网络活动的体外大鼠原代神经元的结果进行了比较。
在多个时间点评估了体外发育神经元网络的基因表达和电活动。分析了 7、28 和 50 天体外的转录组，并与人类脑组织进行了比较。使用多电极阵列（MEA）进行电活动测量的数据表明，神经元网络随着时间的推移逐渐成熟，尽管 hiPSC 中的发育速度比大鼠原代培养物慢。可以检测到调节谷氨酸能、乙酰胆碱能和 GABA 能途径的神经元活性参考药物对 hiPSC 神经元和大鼠原代神经元网络活性的影响。然而，相比之下，GABA 能反应在 hiPSC 神经元中受到限制。
尽管网络发展较慢且网络活动较低，但 CNS.4U®hiPSC 神经元仍可用于检测神经元网络活动的药物诱导变化，如众所周知的致痫药物（影响甘氨酸受体和 Na 通道等）。然而，观察到 GABA 拮抗剂的敏感性较低。
