Immunoglobulin Activation of Fibroblasts

成纤维细胞的免疫球蛋白激活

• 批准号: 8134752
• 负责人: TERRY J SMITH
• 金额: $ 37.32万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8134752
• 关键词: Adenoviruses; Animal Model; Animals; Antibodies; Antigens; Attenuated; Autoantigens; Autoimmune Diseases; Autologous; B-Lymphocytes; Binding; CD19 gene; CD3 Antigens; CD34 gene; CD8B1 gene; Cell physiology; Cells; Chemotactic Factors; Chimeric Proteins; Clinical; Cloning; Complex; Connective Tissue; Development; Disease; Environmental Risk Factor; Epithelium; Event; Exhibits; Experimental Animal Model; Fatty acid glycerol esters; Female; Fibroblasts; Frequencies; Generations; Genetic; Health; Home environment; Hormones; Human; Hyaluronan; Immune; Immune system; Immunoglobulin G; Immunoglobulins; Implant; Inbred BALB C Mice; Infiltration; Inflammation; Inflammatory; Insulin-Like-Growth Factor I Receptor; Label; Lead; Luciferases; Lymphocyte; Lymphocyte Activation; Mediating; Modeling; Monozygotic Twinning; Monozygotic twins; Mus; Newly Diagnosed; Non-Invasive Cancer Detection; Ocular orbit; Orbital implants; Pathogenesis; Patients; Phenotype; Play; Pre-Clinical Model; Process; Production; Proteins; Receptor Activation; Role; Series; Signal Transduction; Staging; Syndrome; T memory cell; T-Cell Receptor; T-Lymphocyte; Testing; Thyroid Diseases; Thyroid Gland; Thyroid Hormones; Thyrotropin Receptor; Tissues; Vision; Yeasts; base; charge coupled device camera; cytokine; disease mechanisms study; effective therapy; in vivo; insight; mouse model; mutant; new therapeutic target; patient population; pre-clinical; receptor; thyroid associated ophthalmopathies; yeast two hybrid system

DESCRIPTION (provided by applicant): Graves' disease (GD) is a syndrome caused by as yet unidentified genetic and environmental factors where the thyroid becomes overactive because antibodies (TSI) directed against the thyrotropin receptor (TSHR) drive excess hormone production. GD is also associated with the inflammation, accumulation of hyaluronan (HA), and remodeling of orbital connective tissue. This process is known as thyroid-associated ophthalmopathy (TAO) for which no effective treatment currently exists. A major impediment to our understanding of TAO is the absence of a pre-clinical animal model. We have discovered previously that orbital fibroblasts from patients with TAO exhibit a unique inflammatory phenotype and over-express the insulin-like growth factor-1 receptor (IGF-1R). IGF-1R-activating antibodies (GD-IgGs) are found in virtually all patients with GD. When they bind to this receptor, T cell chemoattractants and HA are synthesized by TAO fibroblasts. We hypothesize that these events underlie lymphocyte infiltration and expansion of the orbital tissue. We have now discovered that GD is also associated with an increased frequency of IGF-1R+ T and B cells and this phenotype conveys a proliferative and survival advantage. Monozygotic (MZ) twins discordant for GD are also discordant for the IGF- 1R+ lymphocyte skew, suggesting an environmental cause. We have also now discovered that patients with GD have increased numbers of circulating CD34+ fibroblast precursors called fibrocytes. Moreover we find fibrocytes infiltrating the TAO orbit and expressing high levels of TSHR and IGF-1R. In orbital fibroblasts and thyroid epithelium, TSHR and IGF-1R form a physical complex and IGF-1R mediates aspects of TSHR signaling. We have succeeded in cloning an adenovirus encoding an IGF-1R1-GFP fusion protein. When female BALB/c mice are immunized with it, they manifest orbital inflammation and produce GD-IgG. We have also implanted TAO fat into SCID NOD3-/- mice and shown that TAO fibrocytes home to that tissue. We hypothesize that IGF-1R represents the second critical auto-antigen in GD and is necessary with TSHR to generate authentic disease. We now propose studies to test our central hypothesis. Specific aim 1) To characterize profiles of CD4+ IGF-1R+ T cells, CD19+IGF-1R+ B cells, and fibrocytes from patients with GD, determine whether they change longitudinally or predict development of TAO, determine whether IGF-1R+ B cells selectively produce pathogenic Abs such as TSI and GD-IgG, and examine additional MZ twins discordant for clinical GD. Specific aim 2) To characterize the interaction between IGF-1R and TSHR by cloning IGF-1R2 mutants and performing yeast two-hybrid analysis. Specific aim 3) To continue exploiting two mouse models of GD created by 1) implanting orbital fat into SCID NOD3-/- mice and 2) double-immunizing mice with adenoviruses encoding IGF-1R and TSHR. We believe that important insights into disease pathogenesis and identification of novel therapeutic targets will emerge from these studies. PUBLIC HEALTH RELEVANCE: Thyroid-associated ophthalmopathy (TAO) is the sight-threatening orbital component of Graves' disease, a syndrome where the thyroid gland becomes over-active. There are currently no effective therapies for TAO because we lack an experimental animal model to study disease mechanisms and test therapies. In this proposal, we identify abnormalities in human immune system that underlie development of TAO, and apply these insights to animal models in which treatments can be developed.

描述（由申请人提供）：Graves疾病（GD）是一种综合征，是由尚未确定的遗传和环境因素引起的，甲状腺变得过度活跃，因为抗体（TSI）针对甲状腺激素受体（TSHR）驱动过量激素的产生。 GD还与炎症，透明质酸（HA）的积累以及轨道结缔组织的重塑有关。该过程被称为甲状腺相关的眼科（TAO），目前尚无有效的治疗。我们对道的理解的主要障碍是缺乏临床前动物模型。我们之前已经发现，TAO患者的轨道成纤维细胞表现出独特的炎症表型，并表达胰岛素样生长因子-1受体（IGF-1R）。 IGF-1R激活抗体（GD-IGG）几乎在所有GD患者中都发现。当它们与该受体结合时，T细胞化学吸引剂和HA由TAO成纤维细胞合成。我们假设这些事件是淋巴细胞浸润和轨道组织膨胀的基础。现在，我们发现GD还与IGF-1R+ T和B细胞的频率增加有关，并且该表型传达了增殖和生存优势。 GD不一致的单卵（MZ）双胞胎也与IGF-1R+淋巴细胞偏斜不一致，这表明环境原因。我们现在还发现，GD患者的循环CD34+成纤维细胞前体数量增加，称为纤维细胞。此外，我们发现纤维细胞渗入道轨道并表达高水平的TSHR和IGF-1R。在轨道成纤维细胞和甲状腺上皮中，TSHR和IGF-1R形成物理复合物，IGF-1R介导了TSHR信号的各个方面。我们成功地克隆了编码IGF-1R1-GFP融合蛋白的腺病毒。当雌性BALB/C小鼠通过它免疫时，它们会表现出轨道炎症并产生GD-IGG。我们还将陶脂肪植入了SCID NOD3 - / - 小鼠中，并表明TAO纤维细胞是该组织的家。我们假设IGF-1R代表GD中的第二个关键自动抗原，而TSHR是必不可少的，以产生正宗疾病。现在，我们提出研究以检验我们的中心假设。具体目的1）表征CD4+ IGF-1R+ T细胞，CD19+ IGF-1R+ B细胞的谱图以及来自GD患者的纤维细胞，确定它们是纵向变化还是预测TAO的发展，请确定IGF-1R+ B细胞是否选择性地产生了诸如TSI和GD-GDIGG的致病性ABS，以及for gd ins for for gdince for for gdins discord cintic cintic clindic cintic clindict twins twince clincord discord。具体目标2）通过克隆IGF-1R2突变体并进行酵母两杂化分析来表征IGF-1R和TSHR之间的相互作用。特定目标3）继续利用由1）将轨道脂肪植入SCID NOD3 - / - 小鼠和2）用编码IGF-1R和TSHR的腺病毒的双重免疫小鼠的GD模型。我们认为，从这些研究中出现了对疾病发病机理和新型治疗靶标的鉴定的重要见解。公共卫生相关性：与甲状腺相关的眼科（TAO）是威胁坟墓疾病的视觉轨道成分，该疾病是一种综合征，甲状腺变得过于活跃。目前尚无对道的有效疗法，因为我们缺乏研究疾病机制和测试疗法的实验动物模型。在此提案中，我们确定了陶的发展基础的人类免疫系统中的异常，并将这些见解应用于可以开发治疗的动物模型。