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 该项目的目标是开发构象特异性的新型生物制剂， 功能性调节参与疼痛信号传导的电压门控钠（Nav）通道。目标1将 使用计算机结构建模来设计电压敏感结构域的稳定表位模拟物 人NaV1.7、NaV1.8和NaV1.9的IV。在目标2中，将这些纯化的蛋白质片段 在大肠杆菌中表达大肠杆菌，纯化，并用于免疫美洲驼和小鼠的纳米抗体（nAb）， 单克隆抗体（mAb）、兔mAb（R-mAb）和单链可变区片段 （scFv）生产。目的3中的实验将分析mAb的药理学活性 将确定异源系统中针对人NaV通道的靶向性和中试体内功效 在用化疗剂奥沙利铂治疗的大鼠中。何塞·马奎兹先生的工作将扩展到 在目标3中概述的研究NaV表达和功能如何被修饰的努力之后， 急性和慢性奥沙利铂治疗后遗传鉴定的伤害感受器， Abs在小鼠中临床前功效的扩展热感觉行为分析。自从加入博士 Theanne Griffith的实验室作为2021年9月的研究生，Marquez先生 获得的经验与几个技术概述的目标3的母基金，并将 在此基础上，我们将解决两个基本问题， 疼痛时钠通道功能障碍：1）奥沙利铂治疗如何改变钠通道 表达和功能的遗传识别伤害感受器，以及2）如何抑制钠 生物制剂（如Abs和nAbs）的通道功能改变奥沙利铂诱导的热痛 行为？他的研究计划与精心制定的职业发展计划相结合， 将马奎兹先生定位为疼痛领域有竞争力的博士后候选人。