Sotelo Jesus, Tabatabaei Sahar Mofidi, Fofie Christian, Fosu Kelvin, Dodd-O Joseph, Simcik Rebecca, Tack See, Soto-Reyes Miguel J, Yousef Saad, Caro-Diaz Eduardo J, Kumar Vivek, Van Horn Wade, Kolber Benedict J, Tidgewell Kevin J
Department of Biological Sciences, Department of Neuroscience, and Center for Advanced Pain Studies, University of Texas at Dallas, Richardson, TX.
Department of Pharmaceutical Sciences, University of Kentucky, Lexington, KY.
bioRxiv. 2025 Jun 22:2025.06.19.660581. doi: 10.1101/2025.06.19.660581.
Ver E's structure was validated by H NMR, HRMS, and molecular networking analyses. Computational docking and NMR titration confirmed direct, saturable, and tight binding of Ver E to the human Sigma-2 receptor/transmembrane protein 97 (σR/TMEM97). Functional calcium imaging in primary mouse sensory neurons revealed that Ver E increases intracellular Ca levels without modulating store-operated calcium entry (SOCE). Multi-well microelectrode array experiments using human induced pluripotent stem cell (hiPSC) derived nociceptors showed that Ver E significantly reduced neuronal activity at physiological temperatures, but not under heat-stress conditions. Ver E exhibited no cytotoxicity at concentrations up to 30 μM in HEK293 cells, and immunocytochemistry confirmed that it does not alter phosphorylated eIF2α (p-eIF2α) expression, indicating a mechanism distinct from integrated stress response modulators. Collectively, these findings position Ver E as a non-toxic compound capable of selectively modulating neuronal excitability, thereby advancing the development of novel therapeutics for pain management.
Ver E的结构通过核磁共振氢谱（¹H NMR）、高分辨质谱（HRMS）和分子网络分析得到验证。计算对接和核磁共振滴定证实了Ver E与人西格玛-2受体/跨膜蛋白97（σR/TMEM97）直接、可饱和且紧密的结合。原代小鼠感觉神经元的功能性钙成像显示，Ver E可增加细胞内钙水平，而不调节储存式钙内流（SOCE）。使用人诱导多能干细胞（hiPSC）衍生的伤害感受器进行的多孔微电极阵列实验表明，Ver E在生理温度下显著降低神经元活性，但在热应激条件下则不然。在HEK293细胞中，浓度高达30 μM时Ver E均未表现出细胞毒性，免疫细胞化学证实其不会改变磷酸化真核起始因子2α（p-eIF2α）的表达，表明其作用机制不同于整合应激反应调节剂。总体而言，这些发现表明Ver E是一种能够选择性调节神经元兴奋性的无毒化合物，从而推动了新型疼痛管理疗法的开发。
