Investigating the contributions of voltage gated sodium channels to oxaliplatin induced neuropathy

项目总结 该项目的目标是开发特定构象的新型生物制品，以靶向和 功能调制参与疼痛信号的电压门控钠(NAV)通道。目标1将 在硅结构模型中用于为电压敏感领域设计稳定的表位模拟 人类NAV1.7、NAV1.8和NAV1.9的IV。在目标2中，这些纯化的蛋白质片段将被 在大肠杆菌中表达，纯化，并用于免疫骆驼和小鼠的纳米体(NAB)， 单抗(MAbbs)、兔单抗(R-mAbbs)和单链可变片段 (ScFv)生产。目标3中的实验将分析单抗的药理活性。 将确定针对异种系统中的人NAV通道和体内试验的有效性 接受化疗药物奥沙利铂治疗的大鼠。何塞·马奎斯先生的工作将扩大 根据目标3中概述的努力，研究NAV的表达和功能是如何在 急性和慢性奥沙利铂治疗后基因识别的伤害性感受器，以及 单抗临床前疗效的扩展热敏行为学分析。自从加入Dr。 2021年9月作为研究生的Theanne Griffith的实验室，Marquez先生 获得了父母资助的目标3中概述的几种技术的经验，并将 在此基础上，威尔将解决两个关于 疼痛时钠通道功能障碍：1)奥沙利铂治疗如何改变钠通道 在基因可识别的伤害性感受器中的表达和功能，以及2)抑制钠是如何 生物制剂如单抗和NaBS对奥沙利铂引起的热痛的影响 行为？他的研究计划与精心制定的职业发展计划相结合，将 让马奎兹先生成为疼痛领域一名有竞争力的博士后候选人。

PROJECT SUMMARY The goal of this project is to develop conformationally-specific novel biologicals to target and functionally modulate voltage-gated sodium (Nav) channels involved in pain signaling. Aim 1 will use in silico structural modeling to design stable epitope mimetics for the voltage-sensing domain IV of human NaV1.7, NaV1.8, and NaV1.9. In Aim 2, these purified protein fragments will be expressed in E. coli, purified, and used to immunize llamas and mice for nanobody (nAb), monoclonal antibodies (mAbs), rabbit-mAbs (R-mAbs), and single-chain variable fragments (scFvs) production. Experiments in Aim 3 will analyze the pharmacological activity of mAbs against human NaV channels in heterologous systems and pilot in vivo efficacy will be determined in rats treated with the chemotherapeutic agent, oxaliplatin. Mr. Jose Marquez’s work will expand upon the efforts outlined in Aim 3 to investigate how NaV expression and function is modified in genetically identified nociceptors following acute and chronic oxaliplatin treatment, as well as an expanded thermosensory behavioral analysis of Abs preclinical efficacy in mice. Since joining Dr. Theanne Griffith’s laboratory as a graduate student in September of 2021, Mr. Marquez has gained experience with several of the techniques outlined in Aim 3 of the parent grant and will build upon this foundation by will addressing two basic questions regarding mechanisms of sodium channel dysfunction during pain: 1) How does oxaliplatin treatment alter sodium channel expression and function in genetically identifiable nociceptors, and 2) How does inhibiting sodium channel function with biologics, such as Abs and nAbs, alter oxaliplatin induced thermal pain behaviors? His research plan in combination with a carefully crafted career development plan will position Mr. Marquez to be a competitive postdoctoral candidate in the pain field.