scFv Antibody Therapy for Chronic Trigeminal Neuropathic Pain

scFv 抗体治疗慢性三叉神经病理性疼痛

• 批准号: 9912752
• 负责人: Adinarayana Kunamneni
• 金额: $ 18.96万
• 依托单位: UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9912752
• 关键词: Acute; Address; Analgesics; Animal Model; Antibodies; Antibody Therapy; Anticonvulsants; Antidepressive Agents; Anxiety; Autopsy; Behavior; Behavior assessment; Behavioral; Binding; Biodistribution; Blocking Antibodies; Blood; Brain; CCKBR gene; Cells; Chicago; Cholecystokinin; Cholecystokinin B Receptor; Chronic; Clinical; Clinical Trials; Cognitive; Corpus striatum structure; Data; Development; Dose; Effectiveness; Experimental Models; Face; Facial Injuries; Female; Free Ribosome; Future; Gene Expression; Gene Expression Profile; Genes; Goals; Grant; Histopathology; Human; Hypersensitivity; Immune; Immune response; Immunize; Inflammation Mediators; Injury; Legal patent; Light; Literature; Mechanics; Medial; Modeling; Mus; Nerve; Neuroglia; Neurons; Neuropathy; Nociception; Opioid; Orofacial Pain; Outcome; Pain; Pain management; Patients; Peptides; Phase; Physical Function; Post-Traumatic Stress Disorders; Posterior Horn Cells; Preclinical Testing; Prefrontal Cortex; Receptor Activation; Reflex action; Serum; Signal Pathway; Site; Source; Spinal; Structure of trigeminal ganglion; Symptoms; Technology; Testing; Therapeutic; Time; Traumatic injury; Trigeminal System; Trigeminal nerve structure; Universities; anxiety-related behavior; base; behavior measurement; brain circuitry; brain dysfunction; cell type; central sensitization; chronic pain; data submission; effective therapy; effectiveness evaluation; efficacy testing; experience; extracellular; gender difference; in vivo evaluation; in vivo imaging; in vivo imaging system; male; mental function; microchip; mouse model; multidisciplinary; nerve injury; neuropathology; non-opioid analgesic; novel; opiate tolerance; orofacial; pain behavior; pain inhibition; pain model; pain relief; pain signal; painful neuropathy; preclinical study; preference; prevent; receptor; response; social stress; therapeutic development; transmission process; vehicular accident

Abstract Chronic orofacial neuropathic pain that persists after motor vehicle accidents and other facial injuries can be excruciating, unrelenting, and a difficult clinical challenge. Sensitized nerves in the face overactivate not only the trigeminal ganglia neurons but also the second order trigeminal spinal V dorsal horn neurons and glia. Continued activation of pain transmission circuitry results in central sensitization and can exert powerful dysfunctional influences at brain sites associated with anxiety and PTSD. Chronic pain can induce permanent brain circuitry alterations that further diminish physical and mental function. Current treatment with analgesics combined with antidepressants and/or anticonvulsants are generally unsatisfactory in providing pain relief. While opiates are heavily overprescribed to the point of national crisis, they are of little therapeutic value for treatment of orofacial pain. Effective non-addictive, non-opioid therapeutics for chronic orofacial pain remain a critical need. Experimental models in which pain signaling pathways are chronically activated by mild traumatic injury to the trigeminal nerve offer a compelling avenue for discovery and development of new non-addictive, non-opioid therapies. In our trigeminal neuropathic orofacial pain mouse model the trigeminal nerve is compressed to mimic post-traumatic orofacial neuropathic pain in patients. Our recent gene profiling revealed significant 4.0- and 2.7- fold increases in cholecystokinin B (CCK-B) receptor gene expression in the trigeminal ganglia at 3 and 21 days, respectively. The proposed collaborative, multidisciplinary studies will develop novel, brain-penetrant single- chain Fragment variable (scFv) antibody therapies to block CCK-B receptor activation. Aim 1 proposes to generate a repertoire of scFv antibodies using cell-free ribosome display technology to develop, characterize, and authenticate a panel of antibodies to block CCK-B receptor activation. The Aim 2 studies propose testing optimal CCK-B receptor scFv antibodies as therapy to alleviate reflexive and cognitive dependent pain- and anxiety-related behaviors in our orofacial pain model. Preliminary data demonstrate the effectiveness of a single dose of CCK-B receptor scFv antibody administered 3 weeks after model induction during the transition from acute to chronic pain. The proposed studies will further test efficacy of scFv antibody therapy at 8 weeks when hypersensitivity is chronic. Addition of a fluorescent tag on the scFv antibodies will allow additional experimental read-outs including biodistribution with in vivo imaging and postmortem neuropathology to characterize the neuronal and glial cell types involved and their localization. Hypothesis: scFv antibody block of CCK-B receptors is effective therapy for chronic orofacial pain. Aim 1. Generate a panel of CCK-B receptor-specific single-chain Fragment variable (scFv) antibodies using cell-free ribosome display technology to develop/characterize/authenticate a pain therapeutic. Aim 2. Evaluate the effectiveness of CCK-B receptor scFv antibody therapy in male and female mice with acute to chronic and chronic trigeminal nerve injury-induced pain- and anxiety-related behaviors.

摘要 在机动车事故和其他面部损伤后持续存在的慢性口面神经性疼痛， 折磨人的，无情的，困难的临床挑战。面部的敏感神经不仅会过度激活 三叉神经节神经元以及三叉神经脊V背角神经元和胶质细胞。继续 疼痛传递回路的激活导致中枢敏感化， 影响了大脑中与焦虑和创伤后应激障碍相关的部位。慢性疼痛可以诱发永久性的大脑回路 进一步削弱身体和精神功能的改变。目前使用镇痛药联合 抗抑郁药和/或抗惊厥药在提供疼痛缓解方面通常不能令人满意。虽然鸦片是 由于严重过量使用，导致国家危机，因此它们对于治疗口腔面部疾病几乎没有治疗价值 痛苦有效的非成瘾性，非阿片类药物治疗慢性口面疼痛仍然是一个关键的需求。 实验模型中，疼痛信号通路被轻度创伤性损伤慢性激活， 三叉神经为发现和开发新非成瘾性、非阿片类药物提供了一条引人注目的途径 治疗在我们的三叉神经性口面疼痛小鼠模型中，三叉神经被压迫以模拟 创伤后口面神经病理性疼痛的患者。我们最近的基因分析显示，4.0和2.7- 在第3天和第21天，三叉神经节中胆囊收缩素B（CCK-B）受体基因表达成倍增加， 分别拟议的合作，多学科研究将开发新的，大脑渗透的单一， 链可变片段（scFv）抗体疗法来阻断CCK-B受体活化。目标1建议， 使用无细胞核糖体展示技术产生scFv抗体库，以开发、表征 并鉴定一组抗体以阻断CCK-B受体活化。Aim 2研究提出了测试 最佳CCK-B受体scFv抗体作为缓解反射性和认知依赖性疼痛的疗法-以及 焦虑相关的行为在我们的口面疼痛模型。初步数据表明， 在模型诱导后3周，在从CCK-B受体scFv抗体向CCK-B受体scFv抗体的过渡期间，给予一定剂量的CCK-B受体scFv抗体。 急性至慢性疼痛。所提出的研究将在8周时进一步测试scFv抗体疗法的功效， 过敏是慢性的。在scFv抗体上添加荧光标签将允许额外的实验性抗体。 包括生物分布的读数，体内成像和死后神经病理学，以表征 涉及的神经元和神经胶质细胞类型及其定位。 假设：CCK-B受体的scFv抗体阻断是治疗慢性口面部疼痛的有效方法。 目标1。生成一组CCK-B受体特异性单链可变片段（scFv）抗体 使用无细胞核糖体展示技术来开发/表征/鉴定疼痛治疗剂。 目标二。评价CCK-B受体scFv抗体治疗雄性和雌性小鼠的有效性， 急性至慢性和慢性三叉神经损伤引起的疼痛和焦虑相关行为。