Tr1-Specific Tolerance: a Novel Treatment of Multiple Sclerosis

Tr1 特异性耐受：多发性硬化症的新疗法

• 批准号: 8195653
• 负责人: Kanneganti Murthy
• 金额: $ 31.95万
• 依托单位: RADIKAL THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8195653
• 关键词: Acute; Address; Adoptive Transfer; Aftercare; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Antigens; Autoantigens; Autoimmune Diseases; Binding Sites; Bypass; CD28 gene; CD4 Positive T Lymphocytes; CD46 Antigen; CXC chemokine receptor 3; CXCL10 gene; CXCL11 gene; CXCL9 gene; CXCR3 gene; Calcineurin inhibitor; Cell physiology; Cells; Cellular Immunity; Chimeric Proteins; Clinical; Control Groups; Defect; Deterioration; Dexamethasone; Disease; Equilibrium; Etiology; Experimental Autoimmune Encephalomyelitis; FDA approved; Functional disorder; Grant; Histologic; Homologous Gene; Human; IL2RA gene; IgG1; Immune Tolerance; Immunity; Immunosuppressive Agents; In Vitro; Incubated; Inflammation; Inflammatory; Inflammatory Response; Injury; Interleukin-10; Ligand Binding; Location; Mediating; Mitoxantrone; Modeling; Monitor; Multiple Sclerosis; Mus; Myelin; Neurologic; Neurons; Onset of illness; Paralysed; Pathway interactions; Patients; Pharmacodynamics; Phase; Phenotype; Phosphorylation; Primates; Production; Property; Receptor Signaling; Recombinant Proteins; Regulatory T-Lymphocyte; Relapse; Relapsing-Remitting Multiple Sclerosis; Relative (related person); Research Design; STAT3 gene; Safety; Severity of illness; Signal Transduction; Sirolimus; Spinal Cord; Spleen; Steroids; T-Lymphocyte; Testing; Th1 Cells; Therapeutic; Tissues; Titrations; Transgenic Organisms; Work; chemokine; copolymer 1; human FRAP1 protein; in vivo; innovation; interferon therapy; lymph nodes; mouse model; natalizumab; novel; receptor; response; transcription factor

DESCRIPTION (provided by applicant): We are developing a novel immunomodulatory therapy, hR-411, to treat relapsing-remitting multiple sclerosis (RRMS). hR-411 restores regulatory T-cell balance by redirecting pathogenic Th1/Th17 cells into tolerance- inducing Tr1 cells. hR-411 is formed from the fusion of human Ig-Fc and CXCL11, a CXCR3-binding ligand that has been recently identified as a counter-regulatory chemokine. In contrast to the pro-inflammatory CXCR3- binding ligands CXCL9 and CXCL10, in vitro exposure of inflammatory effector Th1 and Th17 cells to CXCL11 redirects their polarization into anti-inflammatory Tr1 (FOXp3-CD25-IL-10high) cells. mR-411, the murine homologue of hR-411, profoundly suppresses the neurological and histologic findings of murine EAE, even when initiated after disease onset. The durability of this repolarization is shown in adoptive transfer studies wherein Ag-specific effector Th1 cells isolated from murine EAE donors treated in vivo with mR-411 suppress EAE in recipients with active disease. In contrast to general immunosupresants (steroids, rapamycin, calcineurin inhibitors), mR-411 induces tolerance that is Ag-specific for active disease yet preserves historical cell-mediated immunity to unrelated Ag's. Because CXCL11 interacts via the CXCR3 receptor, it is expected to bypass the CD46 defect in CD4+ cells in MS and fully activate IL-10 expression and induce Tr1 cell polarization. In support of this assumption, CXCL11 strongly induces IL-10 expression in human CD4+ cells co-incubated in vitro with anti-CD28. The purpose of this grant is to establish the pharmacodyamic profile of mR-411 in a classic murine EAE model of relapsing MS ("R-EAE"). The scientific hypothesis to be tested is that mR-411 decreases R-EAE by inhibiting the activation and differentiation of autoantigen-specific pro-inflammatory Th1/Th17 responses predominantly by promoting the activation of Tr1 cell function. Aim #1: Establish a pharmacodynamic profile of mR-411 in a murine model of R-EAE A titration of mR-411 (1, 3, or 10 mg/kg QOD IP) or an irrelevant IgG1 control will be tested in R-EAE, with groups of mice receiving treatment after the acute phase of disease (16-21 days). These treatment groups will be compared to a negative (sham) control group not exposed to PLP139-151, and a positive control group treated with dexamethasone. Animals will be monitored for overt neurological deterioration over a period of 1 month. Spinal cord tissue will be examined for histologic evidence of inflammation and tissue injury. Aim #2: Determination of the in vivo mechanism by which mR-411 decreases disease severity in R-EAE We will test the working hypothesis that mR-411 treatment decreases EAE disease severity by inhibiting the activation of autoantigen-specific Th1/Th17 effector responses. The same experimental paradigm will be employed as in Aim #1. We wil determine how mR-411 treatment alters the number, phenotype, induction, and function of CD4+ Th1/Th17 cells present within the CNS, spleen, and draining lymph nodes following treatment. These studies will use a combination of actively induced and transfer models of EAE employing myelin-specific 5B6 PLP139-151-specific TCR transgenic T cells. PUBLIC HEALTH RELEVANCE: The proposed studies are designed to determine a putative mechanism by which treatment of mice with mR-411, a counter-regulatory tolerance-inducing chemokine fusion protein, may augment regulatory T cell function, thereby specifically suppressing the activity of autoreactive Th1/17 T cells in a mouse model of the relapsing remitting form of multiple sclerosis. This work has important implications for the etiology and treatment of multiple sclerosis.

描述（由申请人提供）： 我们正在开发一种新型免疫调节疗法 hR-411，用于治疗复发缓解型多发性硬化症 (RRMS)。 hR-411 通过将致病性 Th1/Th17 细胞重定向为耐受诱导的 Tr1 细胞来恢复调节性 T 细胞平衡。 hR-411 由人 Ig-Fc 和 CXCL11 融合而成，CXCL11 是一种 CXCR3 结合配体，最近被鉴定为反调节趋化因子。与促炎 CXCR3 结合配体 CXCL9 和 CXCL10 相比，炎症效应 Th1 和 Th17 细胞在体外暴露于 CXCL11 时，会将其极化重定向为抗炎 Tr1 (FOXp3-CD25-IL-10high) 细胞。 mR-411 是 hR-411 的小鼠同源物，即使在疾病发作后开始，也能深刻抑制小鼠 EAE 的神经学和组织学表现。这种复极化的持久性在过继转移研究中得到了证明，其中从小鼠 EAE 供体中分离出的 Ag 特异性效应 Th1 细胞，在体内用 mR-411 处理后，可抑制患有活动性疾病的受体中的 EAE。与一般免疫抑制剂（类固醇、雷帕霉素、钙调神经磷酸酶抑制剂）相比，mR-411 诱导针对活动性疾病的 Ag 特异性耐受，同时保留对无关 Ag 的历史细胞介导的免疫。由于 CXCL11 通过 CXCR3 受体相互作用，因此有望绕过 MS 中 CD4+ 细胞的 CD46 缺陷，完全激活 IL-10 表达并诱导 Tr1 细胞极化。为了支持这一假设，CXCL11 在体外与抗 CD28 共孵育的人 CD4+ 细胞中强烈诱导 IL-10 表达。本次资助的目的是在复发性 MS 的经典小鼠 EAE 模型（“R-EAE”）中建立 mR-411 的药效学特征。待检验的科学假设是，mR-411 主要通过促进 Tr1 细胞功能的激活来抑制自身抗原特异性促炎 Th1/Th17 反应的激活和分化，从而降低 R-EAE。目标#1：在 R-EAE 小鼠模型中建立 mR-411 的药效学特征 将在 R-EAE 中测试 mR-411（1、3 或 10 mg/kg QOD IP）或不相关 IgG1 对照的滴定，小鼠组在疾病急性期（16-21 天）后接受治疗。这些治疗组将与未暴露于 PLP139-151 的阴性（假）对照组和用地塞米松治疗的阳性对照组进行比较。将在 1 个月的时间内监测动物的明显神经功能恶化。将检查脊髓组织是否有炎症和组织损伤的组织学证据。目标#2：确定 mR-411 降低 R-EAE 疾病严重程度的体内机制 我们将测试以下工作假设：mR-411 治疗通过抑制自身抗原特异性 Th1/Th17 效应反应的激活来降低 EAE 疾病严重程度。将采用与目标#1 相同的实验范例。我们将确定 mR-411 治疗如何改变治疗后 CNS、脾脏和引流淋巴结内存在的 CD4+ Th1/Th17 细胞的数量、表型、诱导和功能。这些研究将结合使用髓磷脂特异性 5B6 PLP139-151 特异性 TCR 转基因 T 细胞主动诱导和转移 EAE 模型。 公共卫生相关性： 拟议的研究旨在确定一种假定机制，通过该机制，用mR-411（一种反调节耐受诱导趋化因子融合蛋白）治疗小鼠，可以增强调节性T细胞功能，从而特异性抑制多发性硬化症复发缓解型小鼠模型中自身反应性Th1/17 T细胞的活性。这项工作对多发性硬化症的病因学和治疗具有重要意义。