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.

项目摘要/摘要 慢性偏头痛使人非常虚弱，人们对此知之甚少，治疗选择也有限。尽管 许多促炎免疫细胞的激活已被证明有助于偏头痛的病理生理， CD3+T淋巴细胞，尤其是免疫抑制调节性T(Treg)细胞在 偏头痛的时代化和逆转仍不清楚。我们将解决此应用程序中的知识差距。 一些偏头痛患者外周血中的Treg细胞数量或功能受损。在一个 慢性偏头痛小鼠模型，反复给予硝酸甘油(NTG)，这是偏头痛的可靠触发因素 患者)不仅诱导了持续的行为敏化，而且CD3+T细胞的数量翻了一番 在没有改变Treg细胞数量的情况下，三叉神经节(TG)，再次表明失去了平衡 在免疫激活和免疫抑制之间。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)和垂体腺苷环化酶激活- 激活多肽(PACAP)，提示TG神经元的敏化。小剂量白介素2(LD-IL2) 优先扩增和激活内源性Treg细胞的治疗完全逆转了慢性 在不改变基本伤害性反应的情况下，偏头痛相关行为敏感化。LD-IL2也有效地 使经NTG处理的小鼠的CGRP和PACAP反应性TG神经元数量减少到对照组水平。 从机制上讲，我们发现外周血中转化生长因子β()和白介素10(IL10)都有表达。 LD-IL2需要信号来逆转慢性偏头痛相关的行为和细胞敏化。 在这一应用中，我们建议检验神经免疫相互作用有助于 慢性偏头痛的发生和缓解，可能是通过调节大脑中三叉神经节神经元的敏感性 三叉神经血管通路。首先，我们将有选择地消耗CD4+或CD8+T细胞，以确定哪种T细胞 亚组(S)有助于慢性头痛相关敏感化的发展和解决。到时候我们会的 用流式细胞术进一步研究CD_4~+和CD_8~+细胞内哪个T细胞亚群(S)被改变 重复NTG。其次，为了进一步阐明Tregs和LD-IL2如何逆转偏头痛的时代化，我们将 检测LD-IL_2对TG和硬脑膜Treg细胞转化生长因子β-1和IL-10的影响。Treg选择性基因敲除 将采用Treg细胞的策略和过继转移来确定Treg细胞是否是主要的 转化生长因子-1和IL-10的来源，介导LD-IL-2的作用。最后，我们将测试重复NTG是否 增加三叉神经节神经元CGRP和/或PACAP的表达。有条件的KO小鼠将被用于 选择性地消除初级传入神经元中的CGRP或PACAP信号。我们会问头痛是否 三叉神经节神经元的时序化完全或部分由CGRP和PACAP信号介导。 总而言之，这项研究的结果不仅将阐明神经免疫相互作用如何调节 偏头痛的时代化和逆转，也有利于基于机制的药物发现。