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疾病（例如顶端牙周炎）方面取得了成功，但仍有180万患者 治疗六个月后可能会经历持续的疼痛。根尖牙周炎是由感染引起的 牙髓导致机械性异常和炎症。持续的牙齿疼痛增加了负担的成本 和镇痛药的摄入量。此外，非甾体类抗炎药（NSAIDS）和阿片类药物提供 长期服用时，不完全缓解疼痛和不良反应，突出了对新治疗的需求 这可以减轻持续性牙齿疼痛的数百万患者。干细胞的研究 镇痛性特性一直在迅速增长，以期发展出新的镇痛药。功效 在神经性疼痛的临床前模型和患者中，间充质干细胞已得到证明 诊断为偏头痛和骨关节炎。初步数据表明，静脉注射人类 顶端乳头（HSCAP）的干细胞完全逆转与顶端牙周炎相关的超敏反应 老鼠。 HSCAP的RNA测序归纳在受感染牙齿上的RNA测序显示出133倍的表达增加 细胞因子巨噬细胞迁移抑制因子（MIF）。此外，MIF受体CD74和CXCR4 三叉神经节中的TRPV1+神经元共定位。此外，重组MIF的局部注入会逆转 与根尖牙周炎有关的高敏性。最后，来自鼠标共同文化的条件培养基 细胞肉芽肿（感染牙齿）和HSCAP减弱辣椒素引起的Ca2+反应 三叉神经神经元，并通过预先治疗MIF抗体抑制。这些数据支持小说 干细胞抗伤害感受的机制通过干细胞的直接作用，这可能是由MIF介导的。 然而，尽管主要数据表明MIF直接抑制三叉神经神经元活性，但神经元 MIF诱导的抗伤害感受的亚型尚不清楚。顶端牙周炎引起的疼痛通常是 报道称为引用，表明由于中心敏化而引起的非伤害性纤维受到参与。 了解这些亚群中的哪一种介导了MIF诱导的抗吸引力将使我们能够发展 针对与根尖牙周炎有关的持续牙齿疼痛的靶向治疗方法。我们将测试中央 假设MIF诱导的三叉神经节神经元活性的抑制是通过伤害和非 - 伤害性神经元。为了检验该假设，我们将1）进行单细胞RT-PCR和免疫组织化学 确定具有不同神经元标记的MIF受体CD74和CXCR4的共表达，2）使用CRE- 重组酶从特定的神经元种群中有条件地敲除MIF受体CD74和CXCR4 体内并评估其对顶端牙周炎模型中MIF抗伤害感受的贡献。这些研究会 不仅可以对介导MIF诱导的抗吸引物的神经元种群提供新的见解，还提供 作为我作为临床医生职业生涯的出色训练工具。