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 慢性偏头痛高度使人衰弱，理解不足，治疗方案有限。虽然 许多促炎性免疫细胞的激活已显示出有助于偏头痛的病理生理学， CD3+ T淋巴细胞的参与，尤其是免疫抑制调节t（Treg）细胞中的参与 偏头痛变化和逆转仍然未知。我们将解决此应用程序中的知识差距。 一些偏头痛患者在外周血中表现出Treg细胞的数值或功能障碍。在 慢性偏头痛的鼠标模型，重复给药硝酸甘油（NTG，偏头痛的可靠触发因素 在患者中）不仅引起持续的行为敏感性，而且使CD3+ T细胞的数量增加了一倍 三叉神经节（TG）而没有改变Treg细胞的数量，再次表明失去平衡 在免疫激活和抑制之间。重复的NTG还增加了TG神经元的数量 可以被神经肽钙蛋白基因相关肽（CGRP）和腺苷酸环化酶环（CGRP）激活 激活多肽（PACAP），表明TG神经元的敏感性。低剂量白介素2（LD-IL2） 治疗，优先扩展和激活内源性Treg细胞，完全逆转了慢性 与偏头痛相关的行为敏感性，而没有改变基本的伤害性反应。 LD-IL2也有效 将NTG处理的小鼠中的CGRP和PACAP响应TG神经元的数量降低到控制水平。 从机械上讲，我们发现外周转化生长因子β（TGF）和白介素10（IL10） LD-IL2需要信号传导才能逆转慢性偏头痛相关的行为和细胞敏感性。 在此应用中，我们建议检验以下假设，即神经免疫相互作用有助于 慢性偏头痛的发展和解决，可能是通过调节TG神经元在 三角血管通路。首先，我们将有选择地替换CD4+或CD8+ T细胞以确定哪个T细胞 子集有助于慢性标头相关敏化的发展和解决。然后我们会 使用流式细胞仪进一步研究CD4+和CD8+细胞中哪种T细胞亚型 重复NTG。其次，为了进一步阐明Tregs和LD-IL2如何逆转偏头痛的变化，我们将 检查LD-IL2是否会在TG和Dura中的Treg细胞中增加TGFβ1和IL10。 Treg选择性基因敲除 将聘请Treg细胞的策略和适应性转移，以​​确定Treg细胞是否为主要 TGF1和IL10的来源介导LD-IL2的作用。最后，我们将测试是否重复NTG 增加TG神经元中的CGRP和/或PACAP胡椒表达。有条件的KO小鼠将使用 选择性地消除初级传入神经元中的CGRP或PACAP信号传导。我们会问是否标题 编年期完全或部分通过TG神经元中的CGRP和PACAP信号传导介导。 总的来说，这项研究的结果不仅会阐明神经免疫相互作用如何调节 偏头痛变化和逆转，但也有助于基于机制的药物发现。