The mechanism of action of Granulocyte Macrophage-Colony Stimulating Factor in an animal model of Multiple Sclerosis

粒细胞巨噬细胞集落刺激因子在多发性硬化症动物模型中的作用机制

• 批准号: 9392704
• 负责人: Benjamin M Segal
• 金额: $ 23.25万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9392704
• 关键词: Adoptive Transfer; Animal Cancer Model; Animal Model; Antigen-Presenting Cells; Antigens; Asthma; Attenuated; Autoimmune Process; Biological; Biological Assay; Blood - brain barrier anatomy; Bone Marrow; C5a anaphylatoxin receptor; CD4 Positive T Lymphocytes; CXCL1 gene; Cell surface; Cells; Chimera organism; Chronic; Clinical; Colony-Stimulating Factor Receptors; Data; Demyelinating Diseases; Demyelinations; Dendritic Cells; Development; Disease; Disease remission; Drug Targeting; Experimental Autoimmune Encephalomyelitis; Frequencies; Glial Fibrillary Acidic Protein; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor Receptors; Growth Factor; IL8RB gene; Immune Sera; Immunocompetent; Immunologic Factors; Immunosuppressive Agents; In Vitro; Inbred Strains Mice; Inflammatory; Laboratory Animals; Lead; Lesion; Lymphocyte; Mediating; Mediator of activation protein; Mononuclear; Multiple Sclerosis; Mus; Myelin; Myelogenous; Myeloid Cells; Neuraxis; Neurologic; Neurologic Deficit; Neutrophil Infiltration; Pathogenesis; Patients; Peptide antibodies; Pharmaceutical Preparations; Pharmacotherapy; Phase; Phase III Clinical Trials; Phenotype; Property; Protocols documentation; Recombinant Granulocyte Colony Stimulating Factor; Relapse; Relapsing-Remitting Multiple Sclerosis; Research; Resistance; Role; Signal Transduction; Suppressor-Effector T-Lymphocytes; T-Lymphocyte; Testing; Transgenes; Transgenic Organisms; Treatment Factor; arginase; base; brain tissue; cellular targeting; central nervous system demyelinating disorder; chemokine; chemokine receptor; clinical remission; cytokine; disability; granulocyte; immunoregulation; migration; monocyte; multiple sclerosis patient; multiple sclerosis treatment; neutralizing antibody; neutrophil; novel; prevent; prophylactic; targeted agent; therapeutic candidate; therapeutic target; transcriptome; young adult

Abstract Multiple sclerosis (MS), an inflammatory demyelinating disease of the central nervous system (CNS), is the most common cause of non-traumatic neurological disability in young adults in the Western Hemisphere. Disease modifying agents (DMA) that deplete lymphocytes, or block their migration to the CNS, have been shown to reduce the frequency of MS relapses in Phase 3 clinical trials. However, none of those drugs are curative and none are effective in all patients. Granulocyte-macrophage colony-stimulating factor (GM-CSF), a myeloid cell growth factor, has emerged as a promising therapeutic target in MS. GM-CSF producing T cells occur at a relatively high frequency in untreated relapsing remitting MS patients, and have been identified in MS brain tissue. Deficiency or neutralization of GM-CSF confers resistance against experimental autoimmune encephalomyelitis (EAE), an animal model of MS. The mechanism of action of GM-CSF in autoimmune demyelination has yet to be definitively demonstrated. In preliminary studies we found that GM-CSF signaling is necessary for the development of chronic neurological deficits in mice with EAE induced by the adoptive transfer of myelin-specific Th17 cells. GM-CSF blockade or deficiency resulted in a decrease in the numbers and percentages of CNS-infiltrating donor T cells and granulocytes, while CD88 (C5a receptor) expressing myeloid cells were enriched. We found that CNS CD88+ monocytes and myeloid dendritic cells (mDC) are poor antigen presenting cells for myelin-reactive T cells. In animal models of cancer and asthma, CD88+ myeloid cells have immunoregulatory properties. Based on these observations, we propose to investigate two potential mechanisms of action of GM-CSF during the effector phase of EAE. In Aim 1, we will test our hypothesis that GM-CSF deficiency accelerates the conversion of pro-inflammatory iNOS+ myeloid cells in EAE infiltrates to an immunosuppressive CD88+ arginase-1+ phenotype. We will compare cell surface marker expression, antigen presenting capacity, cytokine profiles and immunoregulatory properties of GM-CSF receptor deficient (GM-CSFR-/-) versus WT myeloid cells isolated from the CNS of GM-CSFR-/-/ WT  WT mixed bone marrow chimeric mice at peak EAE. We will also determine whether treatment of GM-CSFR-/- adoptive transfer recipients with CD88 antagonists exacerbates EAE and promotes chronic disability. In Aim 2 we will investigate the role of GM-CSF in the recruitment of neutrophils, that mediate blood-brain-barrier breakdown, to the CNS. We will determine if forced CNS expression of CXCL1 (a neutrophil attracting chemokine), or treatment with recombinant granulocyte-colony stimulating factor (G-CSF), prevents EAE remission in GM- CSFR-/- hosts. The proposed research could increase our understanding of the immunopathogenesis of EAE and MS, and eventually lead to the development of novel myeloid cell modulating drugs for the treatment of MS patients who do not respond to lymphocyte-targeting DMA.

抽象的 多发性硬化症（MS）是一种中枢神经系统（CNS）的炎性脱髓鞘疾病，是 西半球的年轻人最常见的非创伤性神经系统障碍的原因。 疾病修饰剂（DMA）复制淋巴细胞或阻止其迁移到中枢神经系统 显示以减少3阶段临床试验中MS继电器的频率。但是，这些药物都不是 治愈性，没有任何患者有效。粒细胞巨噬细胞刺激因子（GM-CSF），A 髓样细胞生长因子已成为MS中有希望的治疗靶标。 GM-CSF产生T细胞 在未处理复发的MS患者中以相对较高的频率发生，并已在 MS脑组织。 GM-CSF的不足或谈判赋予对实验自身免疫性的抵抗力 脑脊髓炎（EAE），MS动物模型。 GM-CSF在自身免疫性中的作用机理 脱髓鞘尚未得到明确的证明。在初步研究中，我们发现GM-CSF信号传导 对于通过自适应引起的EAE的小鼠的慢性神经系统缺陷是必要的 髓磷脂特异性Th17细胞的转移。 GM-CSF桶或缺陷导致数量减少 和CNS浸入供体T细胞和粒细胞的百分比，而表达CD88（C5A受体） 髓样细胞富集。我们发现CNS CD88+单核细胞和髓样树突状细胞（MDC）很差 用于髓磷脂反应性T细胞的抗原细胞。在癌症和哮喘的动物模型中，CD88+髓样 细胞具有免疫调节特性。基于这些观察结果，我们建议研究两个潜力 GM-CSF在EAE效应阶段的作用机理。在AIM 1中，我们将测试我们的假设 GM-CSF缺乏症可以加速EAE浸润中促iNOS+髓样细胞的转化 免疫抑制CD88+精氨酸酶-1+表型。我们将比较细胞表面标记表达， GM-CSF受体缺乏的抗原表现能力，细胞因子谱和免疫调节特性 （GM-CSFR - / - ）与从GM-CSFR的CNS分离的WT髓样细胞 - / - /WTWT混合骨髓 EAE峰值的嵌合小鼠。我们还将确定是否处理GM-CSFR - / - 自适应转移 具有CD88拮抗剂的接受者加剧了EAE并促进慢性残疾。在目标2中，我们将 研究GM-CSF在募集嗜中性粒细胞中的作用，该作用介导了血脑屏障分解， 到CNS。我们将确定CXCL1的强迫CNS表达（中性粒细胞吸引趋化因子）或 用重组粒细胞 - 胶质刺激因子（G-CSF）治疗可防止GM-的EAE缓解 CSFR - / - 主机。拟议的研究可能会增加我们对EAE免疫发病发生的理解 和MS，并最终导致新型髓样细胞调节药物以治疗 不反应淋巴细胞靶向DMA的MS患者。