１細胞単位のDRGニューロンの発火に基づく化合物の痛みおよび作用機序の予測手法の開発

抄録

Cytotoxicity evaluation is used as a method to predict peripheral neuropathy caused by compounds such as anticancer drugs, but there is a need to construct an assessment method that uses neural activity, which appears in the preliminary stage of cytotoxicity. In this study, we aimed to develop a method for predicting peripheral neuropathy induced by compounds based on electrical activities in vitro sensory neurons. In order to acquire electrical activities on a single neuron, rat DRG neurons were cultured on the 236,880 electrode CMOS-MEA, and the response to the TRP channel, which is the main channel of pain, was detected. Based on spontaneous activities at 4 weeks of culture, cell bodies were identified and spontaneous activity patterns at the single cell level were classified into 10 clusters. Evoked responses to TRPV1 agonist capsaicin (1, 10, 100 nM), TRPA1 agonist AITC (1, 10, 100 μM), and TRPM8 agonist menthol (1, 10, 100μM) were detected respectively, and evoked response patterns at single cell level were classified into 12 clusters. As a result of analyzing the relationship between the spontaneous activity pattern and the evoked response pattern for each TRP channel, it was found that the relationship differs depending on the type of TRP channel. The assessment method based on the change in the firing pattern at the single cell level is considered to be effective as a method for predicting peripheral neurotoxicity and the mechanism of action of compounds.