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.

项目概要 该项目的目标是开发具有构象特异性的新型生物制品来靶向和 功能性调节参与疼痛信号传导的电压门控钠 (Nav) 通道。 使用计算机结构建模来设计电压传感域的稳定表位模拟物 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是 在大肠杆菌中表达、纯化并用于对美洲驼和小鼠进行纳米抗体 (nAb) 免疫， 单克隆抗体 (mAb)、兔单克隆抗体 (R-mAb) 和单链可变片段 目标 3 中的实验将分析 mAb 的药理活性。 将确定异源系统中针对人类 NaV 通道和试验体内功效 Jose Marquez 先生的研究工作将在接受化疗药物奥沙利铂治疗的老鼠身上得到扩展。 根据目标 3 中概述的努力，研究 NaV 表达和功能如何在 急性和慢性奥沙利铂治疗后基因鉴定的伤害感受器，以及 自从加入 Dr. 以来，对 Abs 临床前功效进行了扩展热感行为分析。 2021 年 9 月，Marquez 先生作为研究生在 Theanne Griffith 的实验室进行了 获得了母基金目标 3 中概述的几种技术的经验，并将 在此基础上，将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 基因可识别的伤害感受器中的表达和功能，以及 2) 抑制钠如何发挥作用 抗体和 nAb 等生物制剂的通道功能可改变奥沙利铂引起的热痛 他的研究计划与精心制定的职业发展计划相结合会怎样？ 使 Marquez 先生成为疼痛领域有竞争力的博士后候选人。