Evaluation of Trigeminal Ganglia Sensory Neuronal Population/s Mediating MIF-Induced Anti-Nociception in a Model of Apical Periodontitis.

根尖周炎模型中三叉神经节感觉神经元群介导 MIF 诱导的抗伤害感受的评估。

• 批准号: 10822712
• 负责人: Josue De Jesus Murillo
• 金额: $ 5.35万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-12-01 至 2027-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10822712
• 关键词: Absence of pain sensation; Advanced Development; Adverse effects; Afferent Neurons; Aftercare; Analgesics; Animal Model; Antibodies; Apical; Attenuated; Behavioral Assay; Bone Resorption; CXCR4 gene; Capsaicin; Cells; Chronic; Clinical; Coculture Techniques; Data; Degenerative polyarthritis; Dental Pulp; Dental Pulp Exposure; Development; Diagnosis; Disease; Enterobacteria phage P1 Cre recombinase; Evaluation; Female; Fiber; Genetic Transcription; Goals; Hypersensitivity; Immunohistochemistry; In Vitro; Infection; Inflammation; Injections; Intake; Left; Macrophage; Maxilla; Mediating; Mesenchymal Stem Cells; Migraine; Modeling; Mus; Neurons; Nociception; Nociceptors; Non-Steroidal Anti-Inflammatory Agents; Opioid; Pain; Patients; Periapical Granuloma; Periapical Periodontitis; Peripheral; Persistent pain; Pharmaceutical Preparations; Physical Dependence; Population; Pre-Clinical Model; Property; Pruritus; Pulp Canals; Quality of life; Recombinants; Reporting; Research; Reverse Transcriptase Polymerase Chain Reaction; Risk; Scientist; Sorting; Structure of trigeminal ganglion; TRPV1 gene; Testing; Therapeutic; Therapeutic Effect; Tooth structure; Toothache; Touch sensation; Training; Transgenic Mice; Work; antinociception; career; central sensitization; conditional knockout; conditioned place preference; cost; cytokine; dental infection; experience; experimental study; human RNA sequencing; human stem cells; in vitro activity; in vivo; insight; intravenous injection; male; mechanical allodynia; mouse Cre recombinase; novel; orofacial; pain relief; painful neuropathy; prevent; receptor; response; spontaneous pain; stem cells; success; targeted treatment; tongue papilla

Abstract Despite the success of root canal treatments at treating diseases such as apical periodontitis, 1.8 million patients may experience persistent pain six months after treatment. Apical periodontitis is caused by an infection of the dental pulp leading to mechanical allodynia and inflammation. Persistent dental pain increases cost of burden and intake of analgesic drugs. Furthermore, non-steroidal anti-inflammatory drugs (NSAIDs) and opioids provide incomplete pain relief and adverse effects when taken chronically, highlighting the need for a novel treatment that can provide relief to millions of patients experiencing persistent dental pain. The study of stem cells for their analgesic properties has been rapidly growing in hopes of developing a novel class of analgesics. The efficacy of mesenchymal stem cells has been demonstrated in pre-clinical models of neuropathic pain and in patients diagnosed with migraines and osteoarthritis. Preliminary data demonstrates that intravenous injections of human stem cells of the apical papilla (hSCAP) fully reverses hypersensitivity associated with apical periodontitis in mice. RNA sequencing of hSCAP homed to the infected tooth shows a 133-fold increase in the expression of the cytokine Macrophage Migratory Inhibitory Factor (MIF). Additionally, MIF receptors CD74 and CXCR4 colocalize on TRPV1+ neurons in the trigeminal ganglia. Moreover, a local injection of recombinant MIF reverses hypersensitivity associated with apical periodontitis. Lastly, conditioned media from co-cultures of mouse periapical granulomas (the infected tooth) and hSCAP attenuates capsaicin evoked Ca2+ response from trigeminal ganglia neurons and is inhibited with pre-treatment of a MIF antibody. This data supports a novel mechanism for stem cell anti-nociception through the direct effect of stem cells that is possibly mediated by MIF. However, despite primary data suggesting MIF directly inhibits trigeminal ganglia neuronal activity, the neuronal subtypes that MIF-induced anti-nociception are unknown. The pain induced by apical periodontitis is often reported as referred, suggesting the involvement of non-nociceptive fibers due to central sensitization. Understanding which of these subpopulations mediate MIF-induced anti-nociception will allow us to develop targeted treatments for persistent dental pain associated with apical periodontitis. We will test the central hypothesis that MIF-induced inhibition of trigeminal ganglia neuronal activity is mediated by nociceptive and non- nociceptive neurons. To test this hypothesis, we will 1) conduct single cell RT-PCR and immunohistochemistry to determine co-expression of MIF receptors CD74 and CXCR4 with different neuronal markers and 2) use Cre- recombinase to conditionally knockout MIF receptors CD74 and CXCR4 from specific neuronal populations in vivo and evaluate their contributions to MIF anti-nociception in a model of apical periodontitis. These studies will not only provide novel insight into the neuronal populations that mediate MIF-induced anti-nociception but also serve as an excellent training vehicle for my career as a clinician-scientist.

抽象的 尽管根管治疗在治疗根尖周炎等疾病方面取得了成功，但仍有 180 万名患者 治疗后六个月可能会出现持续疼痛。根尖周炎是由牙周组织感染引起的 牙髓导致机械性异常疼痛和炎症。持续的牙痛会增加负担成本 以及服用镇痛药物。此外，非甾体抗炎药 (NSAID) 和阿片类药物可提供 长期服用时疼痛缓解不完全和副作用，凸显了对新治疗方法的需要 这可以缓解数百万遭受持续牙痛的患者。干细胞的研究 镇痛特性已迅速增长，希望开发出一类新型镇痛药。功效 间充质干细胞的作用已在神经性疼痛的临床前模型和患者中得到证实 诊断患有偏头痛和骨关节炎。初步数据表明，静脉注射人体 根尖乳头干细胞（hSCAP）完全逆转与根尖周炎相关的过敏反应 老鼠。对感染牙齿的 hSCAP 进行 RNA 测序显示，hSCAP 的表达增加了 133 倍。 细胞因子巨噬细胞迁移抑制因子（MIF）。此外，MIF 受体 CD74 和 CXCR4 共定位于三叉神经节中的 TRPV1+ 神经元。此外，局部注射重组 MIF 可逆转 与根尖周炎相关的过敏。最后，来自小鼠共培养物的条件培养基 根尖肉芽肿（受感染的牙齿）和 hSCAP 会减弱辣椒素引起的 Ca2+ 反应 三叉神经节神经元，并受到 MIF 抗体预处理的抑制。这个数据支持了小说 干细胞抗伤害感受的机制是通过干细胞的直接作用，可能是由 MIF 介导的。 然而，尽管初步数据表明 MIF 直接抑制三叉神经节神经元活动，但神经元 MIF 诱导的抗伤害感受的亚型尚不清楚。根尖周炎引起的疼痛通常是 据报道，表明由于中枢敏化而涉及非伤害性纤维。 了解这些亚群中的哪一个介导 MIF 诱导的抗伤害感受将使我们能够开发 针对与根尖周炎相关的持续性牙痛的针对性治疗。我们将测试中央 假设 MIF 诱导的三叉神经节神经元活动抑制是由伤害性和非刺激性介导的 伤害感受神经元。为了检验这一假设，我们将 1) 进行单细胞 RT-PCR 和免疫组织化学 确定 MIF 受体 CD74 和 CXCR4 与不同神经元标记的共表达，2) 使用 Cre- 重组酶有条件地敲除特定神经元群体中的 MIF 受体 CD74 和 CXCR4 体内并评估其在根尖牙周炎模型中对 MIF 抗伤害感受的贡献。这些研究将 不仅为介导 MIF 诱导的抗伤害感受的神经元群体提供了新的见解，而且 作为我作为临床医生科学家的职业生涯，这是一个极好的培训工具。