Mechanisms of migraine chronification and reversal

偏头痛的慢性化和逆转机制

• 批准号: 10660758
• 负责人: YUQING CAO
• 金额: $ 46.33万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10660758
• 关键词: Adoptive Transfer; Afferent Neurons; Autoimmune Diseases; Behavioral; CD3 Antigens; CD8-Positive T-Lymphocytes; Calcitonin Gene-Related Peptide; Cells; Cervical; Chronic; Chronic Headaches; Defect; Development; Disease; Dose; Dura Mater; Equilibrium; Exhibits; FOS Protein; Face; Flow Cytometry; Foundations; Functional disorder; Future; Headache; Hypersensitivity; IL2 gene; Image; Immune; Immunohistochemistry; Immunosuppression; Inflammatory; Interleukin-10; Interleukin-2; Knock-out; Knockout Mice; Knowledge; Maintenance; Measures; Mediating; Meningeal; Microglia; Migraine; Modeling; Molecular Target; Mus; Neuroimmune; Neurons; Neuropeptides; Nitroglycerin; Pathogenesis; Pathway interactions; Patients; Peptides; Peripheral; R peptide; Recurrence; Regulatory T-Lymphocyte; Resolution; Signal Pathway; Signal Transduction; Source; Stains; Stimulus; Structure of trigeminal ganglion; Subgroup; T-Lymphocyte; T-Lymphocyte Subsets; TGFB1 gene; Testing; Tissues; Transforming Growth Factor beta; Translational Research; Work; behavioral sensitization; cellular sensitization; cellular targeting; conditional knockout; dorsal horn; drug discovery; experimental study; face skin; functional disability; immune activation; insight; knockout gene; migraine treatment; mouse model; nociceptive response; novel; novel therapeutic intervention; peripheral blood; pituitary adenylate cyclase activating polypeptide; polypeptide; response

PROJECT SUMMARY/ABSTRACT Chronic migraine is highly debilitating, poorly understood, and with limited treatment options. Although the activation of many pro-inflammatory immune cells has been shown to contribute to migraine pathophysiology, the involvement of CD3+ T lymphocytes, especially the immunosuppressive regulatory T (Treg) cells, in migraine chronification and reversal remains unknown. We will address the knowledge gap in this application. Some migraine patients exhibit numerical or functional impairment of Treg cell in the peripheral blood. In a mouse model of chronic migraine, repeated administration of nitroglycerin (NTG, a reliable trigger of migraine in patients) not only induced persistent behavioral sensitization, but also doubled the number of CD3+ T cells in the trigeminal ganglia (TG) without altering the number of Treg cells, again suggesting a loss of balance between immune activation and suppression. Repeated NTG also increased the number of TG neurons that can be activated by neuropeptides calcitonin gene-related peptide (CGRP) and pituitary adenylate cyclase- activating polypeptide (PACAP), indicating the sensitization of TG neurons. Low-dose interleukin-2 (ld-IL2) treatment, which preferentially expands and activates endogenous Treg cells, completely reversed chronic migraine-related behavioral sensitizations without altering basal nociceptive responses. Ld-IL2 also effectively reduced the number of CGRP- and PACAP-responsive TG neurons in NTG-treated mice to the control level. Mechanistically, we found that both peripheral transforming growth factor beta (TGF) and interleukin-10 (IL10) signaling are required for ld-IL2 to reverse chronic migraine-related behavioral and cellular sensitizations. In this application, we propose to test the hypothesis that neuro-immune interactions contribute to the development and resolution of chronic migraine, likely through regulating the sensitivity of TG neurons in the trigeminovascular pathway. First, we will selectively deplete CD4+ or CD8+ T cells to determine which T cell subset(s) contribute to the development and resolution of chronic headache-related sensitizations. We will then use flow cytometry to further investigate which T cell subtype(s) within CD4+ and CD8+ cells are altered by repeated NTG. Secondly, to further elucidate how Tregs and ld-IL2 reverses migraine chronification, we will examine whether ld-IL2 increases TGFβ1 and IL10 in Treg cells in TG and dura. Treg-selective gene knockout strategy and adoptive transfer of Treg cells will be employed to determine whether Treg cells are the main source of TGF1 and IL10 that mediate the effects of ld-IL2. Lastly, we will test whether repeated NTG increases CGRP and/or PACAP peptide expression in TG neurons. Conditional KO mice will be used to selectively eliminate CGRP or PACAP signaling in primary afferent neurons. We will ask whether headache chronification is entirely or partially mediated through CGRP and PACAP signaling in TG neurons. Collectively, results from this study will not only shed light on how neuro-immune interactions regulate migraine chronification and reversal, but also facilitate mechanism-based drug discovery.

项目总结/摘要 慢性偏头痛是高度衰弱的，了解甚少，治疗选择有限。虽然 许多促炎性免疫细胞的活化已显示有助于偏头痛病理生理学， CD 3 + T淋巴细胞，特别是免疫抑制调节性T细胞（Treg）参与了 偏头痛的慢性化和逆转仍然未知。我们将解决这个应用程序中的知识差距。 一些偏头痛患者表现出外周血中Treg细胞的数量或功能损伤。中 慢性偏头痛小鼠模型，重复给予硝酸甘油（NTG，偏头痛的可靠触发剂 患者）不仅诱导了持续的行为敏感化，还使患者体内的CD 3 + T细胞数量增加了一倍。 三叉神经节（TG），而不改变Treg细胞的数量，再次表明失去平衡 免疫激活和抑制之间的联系重复NTG也增加了TG神经元的数量， 可被神经肽降钙素基因相关肽（CGRP）和垂体腺苷酸环化酶激活， 激活多肽（PACAP），表明TG神经元的敏化。低剂量白细胞介素-2（ld-IL 2） 优先扩增和激活内源性Treg细胞的治疗完全逆转了慢性 偏头痛相关的行为敏化，而不改变基础伤害性反应。Ld-IL 2还有效地 将NTG处理小鼠中CGRP和PACAP反应性TG神经元的数量减少到对照水平。 从机制上讲，我们发现外周转化生长因子β（TGF β）和白细胞介素-10（IL 10） 信号传导是ld-IL 2逆转慢性偏头痛相关的行为和细胞敏化所必需的。 在本申请中，我们提出测试神经免疫相互作用有助于 慢性偏头痛的发展和解决，可能通过调节TG神经元的敏感性， 三叉神经血管通路首先，我们将选择性地耗尽CD 4+或CD 8 + T细胞，以确定哪种T细胞 亚组有助于慢性头痛相关致敏的发展和消退。然后我们将 使用流式细胞术进一步研究CD 4+和CD 8+细胞中的哪些T细胞亚型被 重复NTG其次，为了进一步阐明TdR和ld-IL 2如何逆转偏头痛慢性化，我们将 检查ld-IL 2是否增加TG和硬脑膜中Treg细胞中的TGFβ1和IL 10。Treg选择性基因敲除 策略和Treg细胞的过继转移将用于确定Treg细胞是否是主要的免疫调节细胞。 TGF β 1和IL 10的来源，其介导ld-IL 2的作用。最后，我们将测试是否重复NTG 增加TG神经元中CGRP和/或PACAP肽的表达。条件性KO小鼠将用于 选择性消除初级传入神经元中的CGRP或PACAP信号。我们会问是否头痛 在TG神经元中，慢性化完全或部分通过CGRP和PACAP信号传导介导。 总的来说，这项研究的结果不仅揭示了神经免疫相互作用如何调节 偏头痛的慢性化和逆转，而且还促进了基于机制的药物发现。