Novel Antigen-Specific Immunomodulatory Tregitope-based Therapy to Address Autoimmune Pathogenesis in Graves' Disease

基于新型抗原特异性免疫调节 Tregitope 的疗法可解决格雷夫斯病的自身免疫发病机制

基本信息

批准号：
9410057
负责人：
Anne Searls DeGroot
金额：
$ 17.7万
依托单位：
EPIVAX, INC.
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9410057
关键词：
Ablation Address Adverse effects Animal Model Antibodies Antibody Response Antigen Targeting Antigens Autoantibodies Autoantigens Autoimmune Diseases Autoimmune Process Autoimmunity B-Lymphocytes Binding Biological Assay Biological Response Modifier Therapy Bystander Suppression CD4 Antigens CD4 Positive T Lymphocytes Cells Chemicals Child Clinical Clinical Treatment Code Disease Down-Regulation Effector Cell Electroporation Epitopes Eragrostis Evaluation Excision Extracellular Domain Eye diseases FDA approved Formulation Future Gland Glycolates Graves&apos Disease HLA-DR3 Antigen Half-Life Homeostasis Human Hyperthyroidism Hypothyroidism Immune Immune Tolerance Immune response Immunity Immunize Immunoglobulin G Life Link Lymphocyte MHC Class II Genes Mediating Methods Microspheres Modeling Monitor Mus Muscle Operative Surgical Procedures Pathogenesis Patients Peptide Hormones Receptors Peptides Peripheral Peripheral Blood Mononuclear Cell Phase Plasmids Pregnant Women Production Property Radioactive Iodine Recombinants Regulatory T-Lymphocyte Replacement Therapy Research Serum Serum Albumin Splenocyte System T cell response T-Lymphocyte Testing Tetanus Toxoid Thyroid Gland Thyroid Hormones Thyroid stimulating immunoglobulins Thyrotropin Receptor Toxic effect base clinical application extracellular immunoregulation improved in vivo mouse model novel product development receptor binding response

项目摘要

ABSTRACT Significance: Graves’ disease (GD), one of the most prevalent autoimmune diseases, is caused by stimulating autoantibodies (TSAb) to the Thyroid-Stimulating Hormone Receptor (TSHR), resulting in hyperthyroidism. Current methods for managing hyperthyroidism in GD target excess thyroid hormone secretion by ablation or removal of the thyroid or by blocking thyroid hormone synthesis. Each of these therapies is associated with adverse effects, most commonly hypothyroidism requiring life-long replacement therapy. Importantly, none of these approaches address the underlying anti-TSHR CD4 T cell response that contributes to the continued production of TSAb auto-antibodies. Autoimmunity is the result of an imbalance in Teffector:Tregulatory (Teff:Treg) immune-cell homeostasis, and a loss of central or peripheral immune tolerance. T regulatory epitopes (Tregitopes) found in IgG stimulate proliferation and activation of natural and induced Tregs, thereby downregulating a Teff response to self-antigens. These Tregitopes when administered with MHC Class II binding self-antigen peptides, will elicit Antigen- Specific Adaptive Tolerance Induction (ASATI). Hypothesis: Tregitopes, in combination with the target antigen —TSHR CD4 T cell effector peptides — will reduce CD4 T cell activation and T-dependent B cell production of TSAb. Due to the short half-life of peptides in vivo, a practical Tregitope BioTherapeutic (Treg-BT) will need to be administered in a suitable delivery system, that improves its PK/PD properties with minimal toxicity. This project will identify TSHR peptides that stimulate a CD4 effector immune response in PBMC from GD patients, and combine the optimal peptide(s) with Tregitopes in an effective delivery vehicle. In Specific Aim 1, TSHR peptides identified to stimulate a CD4 effector cell response in GD patients will be tested for inhibition by Tregitopes in a TSHR bystander suppression assay. Tregitope peptides with and without TSHR peptides will then be formulated into two Treg-BT delivery vehicles: a) chemically ligated to a recombinant FDA-approved human serum albumin (HSA) or b) formulated into poly(lactic-glycolic acid) (PLGA) microspheres. The Treg-BT will then be tested in the TSHR bystander suppression assay with PBMC from GD patients. In Specific Aim 2, the candidate Treg-BT (HSA- Tregitope + TSHR peptide or Tregitope + TSHR peptides in microspheres, with appropriate controls) will be tested in vivo in a mouse model of GD. HLA-DR3 mice will be genetically immunized with an expression plasmid coding for the extracellular domain of TSHR to generate TSAb. After TSAb response is established, the mice will be treated with the optimized Treg-BT and serum TSAb and CD4 T cell responses will be monitored. Overall Impact: These studies will produce candidate Treg-BT which, combined with TSHR peptides, stimulate a Treg-mediated down-regulation of TSAb autoantibodies, with potential fast-track applicability to the treatment of patients with clinical Graves’ disease.

抽象的意义：坟墓疾病（GD）是最普遍的自身免疫性疾病之一，是由刺激自身抗体（TSAB）到甲状腺刺激激素受体（TSHR），导致甲状腺功能亢进。当前管理GD目标中甲状腺功能亢进症的方法超过甲状腺通过消融或去除甲状腺或通过阻塞甲状腺果烯合成来分泌。每个疗法与不良反应有关，最常见的甲状腺功能减退症需要终身替代疗法。重要的是，这些方法都没有解决基础的抗TSHR CD4 t 细胞反应有助于持续产生TSAB自身抗体。自身免疫性是 TEFFOR失衡的结果：Tregulatory（Teff：Treg）免疫细胞稳态，以及中央的丧失或外周免疫公差。 IgG中发现的T调节表位（Tregitapes）刺激增殖并激活自然和诱导的treg，从而下调了对自我抗原的反应。这些tregitopes用MHC II类结合的自我抗原辣椒给药时，将引起抗原特定的自适应耐度诱导（ASATI）。假设：Tregitapes与目标结合抗原 - TSHR CD4 T细胞效应辣椒 - 将减少CD4 T细胞激活和T依赖性B细胞 TSAB的生产。由于体内的Petides的半衰期短，这是一种实用的Tregitope Biothapeitic （Treg-bt）需要在合适的输送系统中进行管理，以改善其PK/PD属性毒性最小。该项目将识别刺激CD4效应器免疫的TSHR Pepperides GD患者的PBMC反应，并将最佳肽与tregitopes结合在一起送货车。在特定的目标1中，鉴定出来刺激CD4效应细胞反应的TSHR肽 TSHR旁观者抑制测定法中的Tregitopes抑制GD患者。 Tregitope 随后有和没有TSHR肽的肽将被配合为两种Treg-BT递送车：A）化学上与重组FDA批准的人血清相册（HSA）或B）配方聚（PLGA）微球。然后将在TSHR旁观者中测试Treg-BT GD患者对PBMC的抑制测定。在特定的目标2中，候选treg-bt（HSA- Tregitope + TSHR Pepperide或Tregitope + TSHR Pepperides在微球中，并具有适当的对照）在GD的小鼠模型中在体内进行测试。 HLA-DR3小鼠将用表达质粒编码TSHR的细胞外结构域生成TSAB。 TSAB响应后已建立，将用优化的Treg-BT和血清TSAB和CD4 T细胞处理小鼠响应将受到监控。总体影响：这些研究将产生候选treg-bt，这是结合TSHR肽，刺激TSAB自身抗体的TREG介导的下调，并具有潜在的快速轨道适用性适用于治疗临床坟墓疾病患者的治疗。