Department of Electronics, Graduate School of Engineering, Tohoku Institute of Technology, Sendai, Miyagi 982-8577, Japan.
Toxicol Sci. 2022 Jun 28;188(1):131-141. doi: 10.1093/toxsci/kfac045.
Drug-induced peripheral neuropathy occurs as an adverse reaction of chemotherapy. However, a highly accurate method for assessing peripheral neuropathy and pain caused by compounds has not been established. The use of human-induced pluripotent stem cell (hiPSC)-derived sensory neurons does not require animal experiments, and it is considered an effective method that can approach extrapolation to humans. In this study, we evaluated the response to pain-related compounds based on neural activities using in vitro microelectrode array (MEA) measurements in hiPSC-derived sensory neurons. Cultured sensory neurons exhibited gene expression of the Nav1.7, TRPV1, TRPA1, and TRPM8 channels, which are typical pain-related channels. Channel-dependent evoked responses were detected using the TRPV1 agonist capsaicin, a TRPA1 agonist, allyl isothiocyanate (AITC), and TRPM8 agonist menthol. In addition, the firing frequency increased with an increase in temperature from 37°C to 46°C, and temperature sensitivity was observed. In addition, the temperature of the peak firing rate differed among individual neurons. Next, we focused on the increase in cold sensitivity, which is a side effect of the anticancer drug oxaliplatin, and evaluated the response to AITC in the presence and absence of oxaliplatin. The response to AITC increased in the presence of oxaliplatin in a concentration-dependent manner, suggesting that the increased cold sensitivity in humans can be reproduced in cultured hiPSC-derived sensory neurons. The in vitro MEA system using hiPSC-derived sensory neurons is an alternative method to animal experiments, and it is anticipated as a method for evaluating peripheral neuropathy and pain induced by compounds.
药物诱导的周围神经病变是化疗的一种不良反应。然而，尚未建立一种评估化合物引起的周围神经病变和疼痛的高度准确方法。使用人诱导多能干细胞（hiPSC）衍生感觉神经元不需要进行动物实验，并且被认为是一种可以接近外推至人类的有效方法。在这项研究中，我们使用体外微电极阵列（MEA）测量在 hiPSC 衍生感觉神经元中基于神经活动评估与疼痛相关的化合物的反应。培养的感觉神经元表现出 Nav1.7、TRPV1、TRPA1 和 TRPM8 通道的基因表达，这些通道是典型的与疼痛相关的通道。使用 TRPV1 激动剂辣椒素、TRPA1 激动剂丙烯基异硫氰酸酯（AITC）和 TRPM8 激动剂薄荷醇检测通道依赖性诱发电响应。此外，随着温度从 37°C 增加到 46°C，发射频率增加，并且观察到温度敏感性。此外，个体神经元的峰值发射率的温度不同。接下来，我们专注于增加冷敏感性，这是抗癌药物奥沙利铂的副作用，并评估 AITC 在奥沙利铂存在和不存在时的反应。在奥沙利铂存在下，AITC 的反应呈浓度依赖性增加，表明人类中冷敏感性的增加可以在培养的 hiPSC 衍生感觉神经元中重现。使用 hiPSC 衍生感觉神经元的体外 MEA 系统是动物实验的替代方法，并且预计它将成为评估化合物引起的周围神经病变和疼痛的方法。
