Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan.
Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Pharmacology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
Neurotoxicology. 2023 May;96:118-128. doi: 10.1016/j.neuro.2023.04.007. Epub 2023 Apr 20.
Chemotherapy-induced peripheral neuropathy (CIPN) is a major dose-limiting side effect, with no approved therapy for prevention or treatment. Here, we aimed to establish a high-content image platform based on the neurite outgrowth of dorsal root ganglia (DRG)-derived neuron cells for the discovery of neuroprotective agents against paclitaxel-induced CIPN. ND7/23 cells, an immortalized hybrid DRG cell line, were maturely differentiated by induction with nerve growth factor and upregulation of intracellular cAMP levels. High-content image analyses of the neurofilament-stained neurite network showed that paclitaxel disrupted the neurite outgrowth of well-differentiated ND7/23 DRG neuron cells, recapitulating characteristic effects of paclitaxel on primary cultured DRG neurons. This process coincided with the upregulated activity of store-operated Ca entry, similar to those found in rodent models of paclitaxel-induced CIPN. The previously identified neuroprotective agents, minoxidil and 8-Br-cyclic adenosine monophosphate ribose (8-Br-cADPR), attenuated the reduction in total neurite outgrowth in paclitaxel-damaged ND7/23 cells. Additionally, the total neurite outgrowth of well-differentiated ND7/23 cells was concentration-dependently reduced by the neurotoxic chemotherapeutic agents, oxaliplatin and bortezomib, but not the less neurotoxic 5-fluorouracil. We demonstrated that high-content analyses of neurite morphology in well-differentiated DRG neuron-derived cells provide an effective, reproducible, and high-throughput strategy for developing therapeutics against CIPN.
化疗诱导的周围神经病（CIPN）是一种主要的剂量限制副作用，目前尚无批准的预防或治疗药物。在这里，我们旨在建立一个基于背根神经节（DRG）源性神经元细胞的突起生长的高内涵图像平台，用于发现预防紫杉醇诱导的 CIPN 的神经保护剂。ND7/23 细胞是一种永生化的杂交 DRG 细胞系，通过神经生长因子诱导和细胞内 cAMP 水平上调来成熟分化。用神经丝染色的突起网络进行高内涵图像分析显示，紫杉醇破坏了分化良好的 ND7/23 DRG 神经元细胞的突起生长，重现了紫杉醇对原代培养的 DRG 神经元的特征性作用。这一过程与储存操作钙内流活性的上调相一致，与紫杉醇诱导的 CIPN 啮齿动物模型中的情况相似。先前鉴定的神经保护剂米诺地尔和 8-Br-环腺苷单磷酸核糖（8-Br-cADPR）减轻了紫杉醇损伤的 ND7/23 细胞中总突起生长减少。此外，神经毒性化疗药物奥沙利铂和硼替佐米浓度依赖性地降低了分化良好的 ND7/23 细胞的总突起生长，但对神经毒性较小的 5-氟尿嘧啶则没有。我们证明，用高内涵方法分析分化良好的 DRG 神经元源性细胞的突起形态为开发 CIPN 治疗药物提供了一种有效、可重复和高通量的策略。
