Abstract：
BACKGROUND: and Purpose: Chemotherapy-induced peripheral neuropathy (CIPN) constitutes a significant health problem due to the increasing prevalence and lack of therapies for treatment and prevention. While pivotal for routine cancer treatment, paclitaxel and vincristine frequently cause CIPN and impact the quality of life among cancer patients and survivors. Here, we investigate molecular mechanisms and drug transport in CIPN.
METHODS: Human sensory neurons were derived from induced pluripotent stem cells (iPSC-SNs), which were characterized using flow cytometry and immunolabeling. These iPSC-SNs were exposed to different concentrations of the two microtubule-targeting agents, paclitaxel and vincristine, with and without pre-exposure to inhibitors and inducers of efflux transporters. Neuronal networks were quantified via fluorescent staining against sensory neuron markers. Transcriptional effects of the chemotherapeutics were examined using quantitative polymerase chain reactions (qPCR).
RESULTS: Paclitaxel exposure resulted in axonal retraction and thickening, while vincristine caused fragmentation and abolishment of axons. Both agents increased the mRNA expression of the pain receptor, transient receptor potential vanilloid (TRPV1), and highly induced neuronal damage, as measured by activating transcription factor 3 (ATF3) mRNA. iPSC-SNs express the efflux transporters, P-glycoprotein (P-gp, encoded by ABCB1) and multidrug resistance-associated protein 1 (MPR1, encoded by ABCC1). Modulation of efflux transporters indicate that P-gp and MRP1 play a role in modulating neuronal accumulation and neurotoxicity in preliminary experiments.
CONCLUSIONS: and Implications: iPSC-SNs are a valuable and robust model to study the role of efflux transporters and other mechanistic targets in CIPN. Efflux transporters may play a role in CIPN pathogenesis as they regulate the disposition of chemotherapy to the peripheral nervous system, and they may present potential therapeutic targets for CIPN.
METHODS: Human sensory neurons were derived from induced pluripotent stem cells (iPSC-SNs), which were characterized using flow cytometry and immunolabeling. These iPSC-SNs were exposed to different concentrations of the two microtubule-targeting agents, paclitaxel and vincristine, with and without pre-exposure to inhibitors and inducers of efflux transporters. Neuronal networks were quantified via fluorescent staining against sensory neuron markers. Transcriptional effects of the chemotherapeutics were examined using quantitative polymerase chain reactions (qPCR).
RESULTS: Paclitaxel exposure resulted in axonal retraction and thickening, while vincristine caused fragmentation and abolishment of axons. Both agents increased the mRNA expression of the pain receptor, transient receptor potential vanilloid (TRPV1), and highly induced neuronal damage, as measured by activating transcription factor 3 (ATF3) mRNA. iPSC-SNs express the efflux transporters, P-glycoprotein (P-gp, encoded by ABCB1) and multidrug resistance-associated protein 1 (MPR1, encoded by ABCC1). Modulation of efflux transporters indicate that P-gp and MRP1 play a role in modulating neuronal accumulation and neurotoxicity in preliminary experiments.
CONCLUSIONS: and Implications: iPSC-SNs are a valuable and robust model to study the role of efflux transporters and other mechanistic targets in CIPN. Efflux transporters may play a role in CIPN pathogenesis as they regulate the disposition of chemotherapy to the peripheral nervous system, and they may present potential therapeutic targets for CIPN.
摘要：
背景：和目的：化疗引起的周围神经病变（CIPN）由于患病率增加和缺乏治疗和预防疗法而构成了重大的健康问题。虽然对于常规癌症治疗至关重要，紫杉醇和长春新碱经常导致CIPN并影响癌症患者和幸存者的生活质量。这里,我们研究了CIPN的分子机制和药物转运。
方法：人类感觉神经元来源于诱导多能干细胞（iPSC-SNs），使用流式细胞术和免疫标记进行表征。这些iPSC-SNs暴露于不同浓度的两种微管靶向剂，紫杉醇和长春新碱,有或没有预先暴露于外排转运蛋白的抑制剂和诱导剂。通过针对感觉神经元标记的荧光染色来量化神经元网络。使用定量聚合酶链反应（qPCR）检查化学疗法的转录作用。
结果：紫杉醇暴露导致轴突收缩和增厚，而长春新碱引起轴突的破碎和消灭。两种药物都增加了疼痛受体的mRNA表达，瞬时受体电位香草酸（TRPV1），和高度诱导的神经元损伤，如通过激活转录因子3(ATF3)mRNA测量的。iPSC-SNs表达外排转运蛋白，P-糖蛋白（P-gp，由ABCB1编码)和多药耐药相关蛋白1(MPR1，由ABCC1编码)。外排转运蛋白的调节表明，在初步实验中，P-gp和MRP1在调节神经元积累和神经毒性中起作用。
结论：和含义：iPSC-SNs是研究外排转运蛋白和其他机制靶标在CIPN中的作用的有价值且稳健的模型。外排转运蛋白可能在CIPN发病机制中发挥作用，因为它们调节化疗对周围神经系统的处置，它们可能为CIPN提供潜在的治疗靶点。
方法：人类感觉神经元来源于诱导多能干细胞（iPSC-SNs），使用流式细胞术和免疫标记进行表征。这些iPSC-SNs暴露于不同浓度的两种微管靶向剂，紫杉醇和长春新碱,有或没有预先暴露于外排转运蛋白的抑制剂和诱导剂。通过针对感觉神经元标记的荧光染色来量化神经元网络。使用定量聚合酶链反应（qPCR）检查化学疗法的转录作用。
结果：紫杉醇暴露导致轴突收缩和增厚，而长春新碱引起轴突的破碎和消灭。两种药物都增加了疼痛受体的mRNA表达，瞬时受体电位香草酸（TRPV1），和高度诱导的神经元损伤，如通过激活转录因子3(ATF3)mRNA测量的。iPSC-SNs表达外排转运蛋白，P-糖蛋白（P-gp，由ABCB1编码)和多药耐药相关蛋白1(MPR1，由ABCC1编码)。外排转运蛋白的调节表明，在初步实验中，P-gp和MRP1在调节神经元积累和神经毒性中起作用。
结论：和含义：iPSC-SNs是研究外排转运蛋白和其他机制靶标在CIPN中的作用的有价值且稳健的模型。外排转运蛋白可能在CIPN发病机制中发挥作用，因为它们调节化疗对周围神经系统的处置，它们可能为CIPN提供潜在的治疗靶点。
