Thyroid Derived Peptide Presentation by HLA-DR in Thyroiditis

HLA-DR 在甲状腺炎中呈现甲状腺衍生肽

• 批准号: 10155462
• 负责人: YARON TOMER
• 金额: $ 39.95万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10155462
• 关键词: Amino Acids; Antigen Presentation; Antigens; Arginine; Autoimmune Responses; Binding; Biological Assay; Blocking Antibodies; Calorimetry; Cells; Complex; Data; Disease; Drug Kinetics; Enzyme-Linked Immunosorbent Assay; Epigenetic Process; Epitopes; FOXP3 gene; Gene Expression; Generations; Genes; Genetic; Goals; Grant; Graves' Disease; HLA-DR Antigens; HLA-DR3 Antigen; Hashimoto Disease; Hormones; In Vitro; Knowledge; Lead; Methods; Monoclonal Antibodies; Mus; Pathogenicity; Patients; Peptide Receptor; Peptides; Peripheral Blood Mononuclear Cell; Pharmaceutical Chemistry; Positioning Attribute; Property; RNA Splicing; Susceptibility Gene; T-Lymphocyte; Testing; Therapeutic; Thyroid Gland; Thyroiditis; Thyrotropin Receptor; Time; Titrations; Validation; Variant; analog; antibody test; autoimmune thyroid disease; base; design; experience; experimental study; genome-wide; humanized mouse; in silico; in vivo; mouse model; multidisciplinary; new therapeutic target; novel therapeutic intervention; novel therapeutics; preclinical development; prevent; programs; response; targeted treatment; translational study

Autoimmune thyroid diseases (AITD), including Graves' disease (GD) and Hashimoto's thyroiditis (HT) are currently treated only symptomatically and not by targeting the mechanisms causing disease. Our long-term goal is to design precision-targeted therapies for AITD by blocking presentation of thyroidal antigens to T-cells. In order to block antigen presentation we are targeting HLA-DRb1-Arg74, which we have previously shown to be the key HLA-DR pocket that presents thyroidal peptides triggering AITD. During the last grant period we made significant progress towards our long-term goals: (1) We identified the key TSHR peptide epitope causing GD; (2) We identified Cepharanthine as a compound that blocks DRb1-Arg74 and prevents AITD in a mouse model; (3) We identified D-peptides that block peptide binding to DRb1-Arg74; (4) Using genome-wide approaches we identified new AITD susceptibility genes (e.g. ARID5B); (5) We identified genetic-epigenetic interactions triggering AITD. Building on the progress made in the previous grant period we propose to develop new therapeutic approaches for AITD by blocking antigen presentation. Our hypothesis is that presentation of pathogenic Tg/TSHR peptides to T-cells within the DRb1-Arg74 pocket we discovered is key to triggering AITD, & that blocking peptide binding to this pocket can be used to treat/prevent AITD. Our specific aims are: Specific Aim 1: We will enhance the potency and efficacy of Cepharanthine in blocking antigen presentation within DRb1-Arg74 by creating modified Cepharanthine analogs (MCA's). We will use in silico methods to design MCA's; MCA's will be confirmed using our uniquely designed ELISA and cell based assays, and using ex vivo and in vivo experiments in “humanized” DR3 mice in which we will induce autoimmune thyroiditis. Specific Aim 2: We will generate monoclonal antibodies (MAb's) targeting the HLA-DRb1-Arg74 – Tg.2098 complex. This approach is based on our findings that presentation of the Tg peptide Tg.2098 by DRb1-Arg74 is the key step in triggering AITD. The MAb's we produce will be screened by our in vitro ELISA and cell based assays, and confirmed ex vivo and in vivo in our DR3 “humanized” mouse model of AITD. Specific Aim 3: We will validate that MCA's & MAb's block antigen presentation in patients with AITD. We will validate MCA's and MAb's by testing their ability to inhibit T cell recall responses to thyroidal antigens in peripheral blood mononuclear cells (PBMC's) isolated from AITD patients that are positive for DRb1-Arg74. In summary, our multidisciplinary translational project builds on the knowledge gained in the previous grant period. Our goal is to pursue preclinical development of novel therapies for AITD based on blocking antigen presentation by HLA-DRb1-Arg74. Our collaborative team has the capacity, experience, & expertise to achieve the aims of our proposal. The main advantage of our therapeutic approach is that it is both selective since only T-cells recognizing pathogenic thyroidal peptides are targeted and personalized since only patients carrying the HLA-DRb1-Arg74 will be treated. Our translational studies will hopefully lead to novel therapies for AITD.

自身免疫性甲状腺疾病（AITD），包括Graves病（GD）和桥本甲状腺炎（HT）， 目前仅通过药物治疗，而不是通过靶向致病机制来治疗。我们的长期 目标是通过阻断甲状腺抗原向T细胞的呈递来设计AITD的精确靶向疗法。 为了阻断抗原呈递，我们靶向HLA-DRb 1-Arg 74，我们先前已经证明， 是呈递触发AITD的甲状腺肽的关键HLA-DR口袋。在上一次拨款期间，我们 在实现我们的长期目标方面取得了重大进展：（1）我们确定了关键TSHR肽表位 （2）我们发现千金藤素是一种阻断DRb 1-Arg 74并预防AITD的化合物， 小鼠模型;（3）我们鉴定了阻断肽与DRb 1-Arg 74结合的D-肽;（4）使用全基因组 方法，我们确定了新的AITD易感基因（如ARID 5 B）;（5）我们确定了遗传-表观遗传 触发AITD的交互。在上一个赠款期取得进展的基础上，我们建议制定 通过阻断抗原呈递来治疗AITD的新方法。我们的假设是 我们发现，在DRb 1-Arg 74口袋内，致病性Tg/TSHR肽对T细胞的作用是触发 AITD，并且与该口袋结合的阻断肽可用于治疗/预防AITD。我们的具体目标是： 具体目标1：提高千金藤素阻断抗原提呈的能力和功效 在DRb 1-Arg 74中通过创建修饰的千金藤素类似物（MCA）。我们将使用计算机模拟方法， 设计MCA; MCA将使用我们独特设计的ELISA和基于细胞的测定法进行确认，并使用 在“人源化”DR 3小鼠中的离体和体内实验，其中我们将诱导自身免疫性甲状腺炎。 具体目标2：我们将产生靶向HLA-DRb 1-Arg 74-Tg的单克隆抗体（MAb's）。 复杂.这种方法是基于我们的发现，即DRb 1-Arg 74对Tg肽Tg.2098的呈递是 触发AITD的关键步骤我们生产的单克隆抗体将通过我们的体外ELISA和基于细胞的 在我们的AITD的DR 3“人源化”小鼠模型中进行了离体和体内确认。 具体目标3：我们将验证MCA和MAb是否阻断AITD患者的抗原呈递。我们将 通过测试MCA和MAb抑制T细胞对甲状腺抗原的回忆反应的能力来验证MCA和MAb。 从AITD患者分离的DRb 1-Arg 74阳性的外周血单核细胞（PBMC）。 总之，我们的多学科翻译项目建立在以前的资助中获得的知识基础上 期我们的目标是寻求基于封闭抗原的AITD新疗法的临床前开发 HLA-DRb 1-Arg 74的表达。我们的合作团队有能力、经验和专业知识来实现 我们提案的目的。我们的治疗方法的主要优点是，它是选择性的，因为只有 识别致病性甲状腺肽的T细胞被靶向和个性化，因为只有携带 将治疗HLA-DRb 1-Arg 74。我们的转化研究将有望为AITD带来新的治疗方法。