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) 炎症性脱髓鞘疾病 西半球年轻人非创伤性神经功能障碍的最常见原因。 疾病修饰剂 (DMA) 可以消耗淋巴细胞或阻止其迁移至中枢神经系统 三期临床试验显示可以降低多发性硬化症复发的频率，但这些药物均未达到预期效果。 粒细胞巨噬细胞集落刺激因子（GM-CSF）是一种对所有患者都有效的药物。 骨髓细胞生长因子已成为产生 GM-CSF 的 T 细胞的一个有前景的治疗靶点。 在未经治疗的复发缓解型多发性硬化症患者中发生频率相对较高，并且已在 MS 脑组织缺乏或中和 GM-CSF 赋予对实验性自身免疫的抵抗力。 脑脊髓炎 (EAE)，MS 的动物模型 GM-CSF 在自身免疫中的作用机制。 脱髓鞘作用尚未得到明确证实。在初步研究中，我们发现 GM-CSF 信号传导。 对于因过继引起的 EAE 小鼠慢性神经功能缺损的发展是必要的 髓磷脂特异性 Th17 细胞的转移阻断或缺陷导致数量减少。 以及 CNS 浸润供体 T 细胞和粒细胞的百分比，而 CD88（C5a 受体）表达 我们发现中枢神经系统 CD88+ 单核细胞和骨髓树突状细胞 (mDC) 富集。 髓磷脂反应性 T 细胞的抗原呈递细胞在癌症和哮喘动物模型中，CD88+ 髓样细胞。 基于这些观察，我们建议研究两种潜在的细胞具有免疫调节特性。 GM-CSF 在 EAE 效应阶段的作用机制 在目标 1 中，我们将检验我们的假设： GM-CSF 缺乏加速 EAE 浸润中促炎 iNOS+ 骨髓细胞的转化 免疫抑制性 CD88+ 精氨酸酶-1+ 表型 我们将比较细胞表面标志物的表达， GM-CSF 受体缺陷的抗原呈递能力、细胞因子谱和免疫调节特性 (GM-CSFR-/-) 与从 GM-CSFR-/-/ WT → WT 混合骨髓中枢神经系统分离的 WT 骨髓细胞相比 我们还将确定是否对处于EAE峰值的嵌合小鼠进行GM-CSFR-/-过继转移治疗。 在目标 2 中，接受 CD88 拮抗剂的患者会加剧 EAE 并导致慢性残疾。 研究 GM-CSF 在中性粒细胞募集中的作用，介导血脑屏障破坏， 我们将确定 CXCL1（中性粒细胞吸引趋化因子）是否强制 CNS 表达，或 重组粒细胞集落刺激因子 (G-CSF) 治疗可防止 GM 患者的 EAE 缓解 CSFR-/- 宿主。拟议的研究可以增加我们对 EAE 免疫发病机制的了解。 和 MS，并最终导致新型骨髓细胞调节药物的开发，用于治疗 对淋巴细胞靶向 DMA 无反应的多发性硬化症患者。