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

研究电压门控钠通道对奥沙利铂诱导的神经病变的影响

• 批准号: 10621059
• 负责人: James S Trimmer
• 金额: $ 2.32万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-21 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10621059
• 关键词: Action Potentials; Acute; Address; Afferent Neurons; Antibodies; Antigens; Applications Grants; Behavioral; Binding; Biological Products; Biology; Biometry; C Fiber; Chemotherapy-induced peripheral neuropathy; Chronic; Circular Dichroism; Clinical; Computer software; Data; Development; Development Plans; Electrophysiology (science); Elements; Epitopes; Escherichia coli; Evaluation; FDA approved; Feedback; Foundations; Functional disorder; Generations; Genetic study; Goals; Grant; Human; Human Genetics; Immunize; Immunoglobulin G; Interdisciplinary Study; Intrathecal Injections; Ion Channel; Kinetics; Laboratories; Llama; Manuals; Mediator of activation protein; Modeling; Molecular Conformation; Molecular Probes; Molecular Target; Monoclonal Antibodies; Mus; Neuropathy; Neurosciences; Nociceptors; Oryctolagus cuniculus; Pain; Pain management; Pharmacology; Phase; Positioning Attribute; Pre-Clinical Model; Procedures; Production; Property; Protein Engineering; Protein Fragment; Proteins; Rattus; Recombinant Antibody; Recombinants; Research; Sampling; Sodium; Sodium Channel; Structural Models; Structure; System; Techniques; Technology; Therapeutic; Therapeutic antibodies; Validation; Vertebral column; Work; base; career development; chemotherapeutic agent; chronic pain management; design; experience; experimental study; extracellular; flexibility; graduate student; immunogenicity; in silico; in vivo; insight; mimetics; nanobodies; novel; novel strategies; novel therapeutics; oxaliplatin; pain behavior; pain model; pain signal; parent grant; polyclonal antibody; pre-clinical; preclinical efficacy; programs; protein folding; rational design; screening; structural biology; success; targeted treatment; therapeutic candidate; therapeutic development; trend; voltage

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.

项目摘要 该项目的目的是开发针对特定构象的新型生物制剂来靶向和 在功能上调节参与疼痛信号传导的电压门控钠（NAV）通道。目标1意志 用于在硅结构建模中设计稳定的表位模拟物用于电压感应域 IV人类NAV1.7，NAV1.8和NAV1.9。在AIM 2中，这些纯化的蛋白质片段将是 在大肠杆菌中表达，纯化并用来使乳lam和小鼠免疫用于纳米机（NAB）， 单克隆抗体（mAb），兔-MAB（R-MAB）和单链变量片段 （SCFV）生产。 AIM 3中的实验将分析mAb的药物活性 将确定异源系统中的人类NAV通道和体内效率的飞行员 在用化学治疗剂奥沙利铂处理的大鼠中。何塞·马克斯（Jose Marquez）先生的工作将扩大 AIM 3中概述的努力研究了如何修改NAV的表达和功能 急性和慢性阿沙利铂治疗后，遗传鉴定的伤害感受器以及 扩展的小鼠ABS临床前效率的热感应行为分析。自加入博士以来 Theanne Griffith的实验室是2021年9月的研究生，Marquez先生 在父母赠款的AIM 3中概述的几种技术获得了经验 通过将两个基本问题解决有关机制的两个基本问题 疼痛期间钠通道功能障碍：1）奥沙利铂治疗如何改变钠通道 一般可识别的伤害感受器的表达和功能，2）如何抑制钠 使用生物制剂（例如ABS和NABS）改变奥沙利铂诱导热疼痛的通道功能 行为？他的研究计划与精心制作的职业发展计划相结合 将Marquez先生定位为疼痛领域中有竞争的博士后候选人。