Thy1 expression, adpogenesis, inflammation and orbital remodeling mechanisms in Thyroid Eye Disease

甲状腺眼病中 Thy1 表达、脂肪生成、炎症和眼眶重塑机制

• 批准号: 9213038
• 负责人: RICHARD P. PHIPPS
• 金额: $ 38.46万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9213038
• 关键词: Adipocytes; Alpha Cell; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Area; Autoantigens; Autoimmune Diseases; Biological Assay; Biological Process; Blindness; Cell Surface Proteins; Cell surface; Cells; Cicatrix; Clustered Regularly Interspaced Short Palindromic Repeats; Complementary DNA; Connective Tissue; Deposition; Development; Diplopia; Disease; Equilibrium; Evidence based treatment; Eye; Eye diseases; Fatty acid glycerol esters; Fc Receptor; Fibroblasts; Gene Expression; Gene Expression Profiling; Genetic Transcription; Goals; Graves' Disease; Homeostasis; Human; Impairment; Inflammation; Inflammatory; Ligands; Link; Lipids; Mediating; Mediator of activation protein; Methods; MicroRNAs; Molecular; Myofibroblast; Ocular orbit; Orbital Diseases; Outcome; Patients; Peroxisome Proliferator-Activated Receptors; Phenotype; Plasmids; Post-Translational Protein Processing; Process; Production; Receptor Activation; Receptor Signaling; Regulation; Reporter; Research; Research Project Grants; Role; Signal Transduction; Small Interfering RNA; Stimulus; Testing; Therapeutic Intervention; Thyroid Gland; Thyrotropin Receptor; Tissues; Transcriptional Activation; Translations; Treatment Efficacy; Vision; Work; cytokine; experimental study; human disease; inhibitor/antagonist; lipid biosynthesis; overexpression; receptor expression; response; thyroid associated ophthalmopathies; transcription factor

The Focus: Thyroid eye disease (TED) is a disfiguring and sight-threatening autoimmune disease that involves inflammation and remodeling of the orbit. TED can present itself with mostly orbital fat (Type 1), orbital connective tissue (Type 2) or a combination of both. Here, we focus on how TED manifests itself as Type 1 disease. The Premise: We know that activated orbital fibroblasts produce high levels of inflammatory cytokines and form either connective tissue myofibroblasts or lipid rich inflammatory adipocytes. Our earlier work showed - that only human orbital fibroblasts that do not express a cell surface protein called Thy1 (Thy1 ) can differentiate into adipocytes when stimulated with an adipogenic medium. And, only those that express Thy1 (Thy1+) differentiate into myofibroblasts when provoked by pro-scarring agents like TGF. Herein, we show that Thy1 presence has a direct role in diminishing orbital fibroblast fat accumulation. For example, deliberate - expression of Thy1 in Thy1 fibroblasts blocks their differentiation to adipocytes. Since adipogenesis requires the activation of the transcription factor PPAR we propose that Thy1 impairs PPAR function. Adipogenesis can also manifest after thyroid stimulating hormone receptor (TSHR) signaling and our studies demonstrate that Thy1 and TSHR are inversely related. Goals: (a) test specific hypotheses about mechanistic and causal roles of Thy1 in TED, (b) determine the molecular mechanisms by which Thy1 expression dampens adipogenesis and inflammatory cytokine production. These mechanisms include [1] regulation of TSHR, [2] PPAR activity and [3] modulation of post-transcriptional gene expression through key microRNAs. Organizing Hypotheses: Thy1 is more than a cell-surface marker in TED, as it is also a functional mediator of fibroblast fate in the disease. Thy1+ orbital fibroblasts have decreased PPAR activity, decreased TSHR, and decreased miR-130a expression, which inhibit the fundamental processes in TED of orbital fibroblast adipogenesis and proinflammatory cytokine production. The proposed experiments will test corollaries of this hypothesis using primary human orbital fibroblasts and tissues; because there is no animal model that provides a consistent phenotype of fat accumulation in the orbit. Specific Aim 1: Test the hypothesis that orbital fibroblast Thy1 has anti-adipogenic and anti-inflammatory mechanisms of action by inhibiting PPAR activity and regulating post-transcriptional gene expression. Specific Aim 2: Test the hypothesis that the mechanism by which Thy1 has its anti-adipogenic and anti-inflammatory effects is through the regulation of TSHR. Impact on the field: Discovering that Thy1 has a major biological function influencing eye disease is a major advance opening new areas for research and therapeutic intervention into the aberrant fat deposition and inflammation associated with TED. The inverse relationship that will be investigated between Thy1 and TSHR is the first demonstration that Thy1 can control TSHR. The results will lead to further research on how targeting of Thy1 and/or its anti-adipogenic mechanism of action could be used in more effective therapeutics.

重点：甲状腺眼病（TED）是一种毁容和视力威胁自身免疫性疾病 涉及注射和重塑轨道。 TED可以大部分呈轨道脂肪（1型），轨道 结缔组织（2型）或两者的组合。在这里，我们关注TED如何表现为类型1 疾病。前提：我们知道活化的轨道成纤维细胞会产生高水平的炎性细胞因子 并形成连接的组织肌纤维细胞或富含脂质的炎症脂肪细胞。我们的早期工作显示 - 只有不表达细胞表面蛋白Thy1（Thy1）的人轨道成纤维细胞才能 当用脂肪生成培养基刺激时，分化为脂肪细胞。而且，只有那些表达thy1的人 （Thy1+）当由TGF等亲施加剂引起时，分化为肌纤维细胞。在此，我们表明 Thy1的存在在减少轨道成纤维细胞脂肪积累中具有直接作用。例如，故意 - Thy1在Thy1成纤维细胞中的表达阻断了它们与脂肪细胞的分化。由于成生成需要 转录因子PPAR的激活我们提出THY1会损害PPAR的功能。脂肪形成 在甲状腺刺激马酮受体（TSHR）信号传导后也可以表现出来，我们的研究表明 THY1和TSHR成反比。目标：（a）测试有关机械和因果角色的特定假设 TED中的thy1，（b）确定thy1表达抑制成生成的分子机制 和炎性细胞因子产生。这些机制包括[1]调节TSHR，[2]PPAR活性 [3]通过关键microRNA调节转录后基因表达。组织假设： Thy1不仅是TED中的细胞表面标记，因为它也是成纤维细胞命运的功能介体 疾病。 Thy1+轨道成纤维细胞的PPAR降低，TSHR降低，MiR-130a降低 表达，抑制轨道成纤维细胞成生的TED和 促炎性细胞因子产生。提出的实验将使用 原发性人类轨道成纤维细胞和组织；因为没有动物模型可以提供一致的 轨道中脂肪积累的表型。特定目的1：检验轨道成纤维细胞Thy1的假设 通过抑制PPAR的活性和调节，抗辅助和抗炎作用机制 转录后基因表达。特定目的2：检验以下假设：Thy1的机制 具有抗辅助和抗炎作用是通过调节TSHR的。对现场的影响： 发现Thy1具有影响眼病的主要生物学功能，这是一个主要的开放新的 研究和治疗干预的领域与异常脂肪沉积和相关的炎症 与Ted。 THY1和TSHR之间将研究的反比关系是第一个演示 Thy1可以控​​制TSHR。结果将导致进一步研究THY1和/或ITS的靶向 抗辅助作用机制可用于更有效的治疗。