Contribution of Nuclear S1P Signaling to Microtubule Targeting Agent-Induced Changes in Transcriptional Activity in Human iPS-SNs

项目摘要/摘要 化疗引起的周围神经病变是一种常见的剂量限制性毒性，可降低治疗 癌症患者生活质量的有效性和影响。这项研究的首要目标是 确定化疗所致周围神经病变的分子基础以支持肿瘤的发展 预防和治疗这种毒性的有针对性的治疗。建议的研究是基于反向翻译的 利用遗传关联发现来推断外周关键通路的药物遗传学方法 神经病。最近的遗传关联和功能验证发现支持鞘氨醇-1的作用- 磷酸盐(S1P)信号在化疗所致神经毒性中的作用，这与以前的研究一致 在啮齿动物模型中。本申请中提出的研究将扩展这些发现，并解决重要的 我们对S1P信号在靶细胞、外周感觉神经元中的毒性的认识存在差距。中环 将被检验的假设是微管对外周感觉神经元S1P信号的调制 靶向药物在其神经毒性中起着关键作用。一种人类诱导的多能干细胞来源 化疗神经毒性的感觉神经元模型(iPS-SNS)将用于所有研究。 将使用药理学和遗传学方法来调节S1P信号和询问 化疗毒性与这一信号通路有关。这三个目标是相辅相成、相辅相成的 S1P的离散函数。第一个目标是调查微管靶向剂是否改变鞘磷脂。 S1P受体在感觉神经元中的代谢，并将特定的S1P受体与细胞骨架变化联系起来。这些研究 在第二个目标中提出的将侧重于S1P受体下游的Rho GTP酶信号，并将 建立S1P信号轴，它对化疗引起的轴突结构和 发展缩放式灯泡。第三个目标将使用scRNA-seq和sc-ATACseq来阐明 紫杉醇诱导iPS-SNs基因表达的改变涉及S1P对染色质可及性的影响。这个 Fingolimod的能力，这是一种针对S1P受体信号的多发性硬化症治疗方法，目前正在 经测试可预防和治疗紫杉醇引起的周围神经病变，以防止 将检查化疗引起的神经毒性。总的来说，这些研究将揭示分子 感觉神经元对微管的反应引起轴突变性的机制 靶向药物和阐明使用Fingolimod进行神经保护的新机制。

PROJECT SUMMARY/ABSTRACT Chemotherapy induced peripheral neuropathy is a common dose-limiting toxicity that can reduce therapeutic effectiveness and impact quality of life for cancer patients. The overarching goal of this research is to determine the molecular basis of chemotherapy-induced peripheral neuropathy to support the development of targeted therapies to prevent and treat this toxicity. The proposed studies are based on a reverse translational pharmacogenetic approach that uses genetic association findings to implicate critical pathways in peripheral neuropathy. Recent genetic association and functional validation findings support a role for sphingosine-1- phosphate (S1P) signaling in chemotherapy-induced neurotoxicity, which are consistent with previous studies in rodent models. The studies proposed in this application will extend these findings and address a significant gap in our knowledge of S1P signaling in target cells for toxicity, peripheral sensory neurons. The central hypothesis that will be tested is that modulation of S1P signaling in peripheral sensory neurons by microtubule targeting agents plays a critical role in their neurotoxicity. A human induced pluripotent stem cell derived sensory neuron model of chemotherapy neurotoxicity (iPS-SNs) will be employed for all studies. Pharmacological and genetic approaches will be used to modulate S1P signaling and interrogate chemotherapy toxicity linked to this signaling pathway. The three aims are complementary and address discrete functions of S1P. The first aim will investigate whether microtubule targeting agents alter sphingolipid metabolism in sensory neurons and will link specific S1P receptors to cytoskeletal changes. The studies proposed in the second aim will focus on Rho GTPase signaling downstream of S1P receptors and will establish the S1P signaling axis that is critical for chemotherapy-induced changes in neurite structure and the development of retraction bulbs. The third aim will use scRNA-seq and sc-ATACseq to elucidate whether paclitaxel-induced changes in gene expression in iPS-SNs involve S1P effects on chromatin accessibility. The ability of fingolimod, a multiple sclerosis therapy that targets S1P receptor signaling and is currently being tested for prevention and treatment of paclitaxel-induced peripheral neuropathy, to protect against chemotherapy-induced neurotoxicity will be examined. Collectively, these studies will reveal molecular mechanisms underlying the axon degeneration that occurs in sensory neurons in response to microtubule targeting agents and elucidate novel mechanisms for neuroprotection with fingolimod.