Nimbalkar Siddharth, Guo Xiufang, Colón Alisha, Jackson Max, Akanda Nesar, Patel Aakash, Grillo Marcella, Hickman James J
Hybrid Systems Lab, University of Central Florida, NanoScience Technology Center, Orlando, FL, United States.
Hesperos Inc., Orlando, FL, United States.
Front Cell Dev Biol. 2023 Mar 1;11:1011145. doi: 10.3389/fcell.2023.1011145. eCollection 2023.
The control of severe or chronic pain has relied heavily on opioids and opioid abuse and addiction have recently become a major global health crisis. Therefore, it is imperative to develop new pain therapeutics which have comparable efficacy for pain suppression but lack of the harmful effects of opioids. Due to the nature of pain, any experiment is undesired even in animals. Recent developments in stem cell technology has enabled the differentiation of nociceptors from human induced pluripotent stem cells. This study sought to establish an functional induced pluripotent stem cells-derived nociceptor culture system integrated with microelectrode arrays for nociceptive drug testing. Nociceptors were differentiated from induced pluripotent stem cells utilizing a modified protocol and a medium was designed to ensure prolonged and stable nociceptor culture. These neurons expressed nociceptor markers as characterized by immunocytochemistry and responded to the exogenous toxin capsaicin and the endogenous neural modulator ATP, as demonstrated with patch clamp electrophysiology. These cells were also integrated with microelectrode arrays for analgesic drug testing to demonstrate their utilization in the preclinical drug screening process. The neural activity was induced by ATP to mimic clinically relevant pathological pain and then the analgesics Lidocaine and the opioid DAMGO were tested individually and both induced immediate silencing of the nociceptive activity. This human-based functional nociceptive system provides a valuable platform for investigating pathological pain and for evaluating effective analgesics in the search of opioid substitutes.
严重或慢性疼痛的控制严重依赖阿片类药物，而阿片类药物的滥用和成瘾最近已成为全球主要的健康危机。因此，开发具有可比的疼痛抑制效果但缺乏阿片类药物有害影响的新型疼痛治疗方法势在必行。由于疼痛的性质，即使在动物身上进行任何实验都是不可取的。干细胞技术的最新进展使得从人诱导多能干细胞分化出伤害感受器成为可能。本研究旨在建立一种与微电极阵列集成的功能性诱导多能干细胞衍生的伤害感受器培养系统，用于伤害感受性药物测试。利用改良方案从诱导多能干细胞中分化出伤害感受器，并设计一种培养基以确保伤害感受器的长期稳定培养。这些神经元通过免疫细胞化学鉴定表达伤害感受器标志物，并如膜片钳电生理学所示，对外源毒素辣椒素和内源性神经调节剂ATP作出反应。这些细胞还与微电极阵列集成用于镇痛药物测试，以证明它们在临床前药物筛选过程中的应用。通过ATP诱导神经活动以模拟临床相关的病理性疼痛，然后分别测试镇痛药利多卡因和阿片类药物DAMGO，两者均立即诱导伤害感受性活动沉默。这种基于人的功能性伤害感受系统为研究病理性疼痛和评估寻找阿片类替代物的有效镇痛药提供了一个有价值的平台。
