Molecular And Immunopathology Of Experimental And Clinical Ocular Diseases

实验和临床眼部疾病的分子和免疫病理学

• 批准号: 8938289
• 负责人: Chi-Chao Chan
• 金额: $ 87.77万
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8938289
• 关键词: AKT1 gene; Affect; Age; Age related macular degeneration; American; Animal Model; Apoptotic; Area; Autophagosome; Biological Models; Blindness; Cell Death; Cells; Choroidal Neovascularization; Chronic; Chronic Childhood Arthritis; Clinic; Clinical; Collaborations; Development; Diagnosis; Diet; Disease; Drusen; Elderly; Endotoxins; Epithelium; Etiology; Exudative age-related macular degeneration; Eye; Eye Neoplasms; Eye diseases; Gender; Genes; Genetic; Genotype; Goals; HIV; Human; Immune response; Immunohistochemistry; In Vitro; Inflammation; Inflammatory; Institutes; Interleukin-10; Interleukin-17; Interleukin-6; Interleukins; Intraocular Lymphoma; Knowledge; Link; Lipids; Liquid substance; Lymphoma; Malignant Neoplasms; Methods; MicroRNAs; Microdissection; Modeling; Molecular; Multiple Sclerosis; Mus; Neoplasms; Oxidative Stress; Pathogenesis; Pathology; Patients; Pattern; Physicians; Play; Posterior Uveitis; Procedures; Publishing; Race; Reporting; Research; Retinal; Retinitis; Reverse Transcriptase Polymerase Chain Reaction; Risk Factors; Role; Sarcoidosis; Single Nucleotide Polymorphism; Small Interfering RNA; Smoking; Specimen; Spottings; Structure of retinal pigment epithelium; Sympathetic Ophthalmia; Syndrome; TGFB1 gene; Techniques; Technology; Therapeutic Agents; Tissues; Translating; United States National Institutes of Health; Universities; Uveitis; Vascular Endothelial Growth Factors; Viral; Von Hippel-Lindau Syndrome; Woman; Work; adeno-associated viral vector; animal tissue; autoimmune uveitis; base; cancer cell; caspase-3; caspase-9; fibrillin-2; geographic atrophy; girls; hemangioblastoma; human tissue; immunopathology; improved; intravitreal injection; macrophage; macula; male; molecular pathology; mouse model; neoplastic; novel therapeutics; photoreceptor degeneration; platelet-derived growth factor C; receptor; sex; therapeutic effectiveness; therapeutic target; tumorigenesis

We identify and topographically localize inflammatory, degenerative, and malignant cells, as well as their products and related molecules, in patient specimens and animal tissues. We analyze these cells and their products mainly by routine pathology, immunohistochemistry, and molecular pathology. The application of cutting-edge technology, such as microdissection combined with molecular techniques (PCR, RT-PCR, genotyping, etc.) allows us to provide a more accurate pathological diagnosis (assessment) and understand the pathogenesis of the disease. These technologies and findings will guide us in selecting the most targeted treatments for patients. We also study the mechanisms of different ocular diseases from various animal models. Using animal models, we can assess the efficacy of novel therapeutic agents for various ocular diseases. In FY2014, we accomplished the following in our research: 1. Molecular Pathology of Age-Related Macular Degeneration (AMD): AMD is the leading cause of irreversible severe central vision loss among the elderly in the world. It is projected that AMD will affect 3 million Americans over the age of 50 by the year 2020. The pathology of AMD is characterized by the accumulation of soft drusen, retinal pigment epithelium (RPE) and photoreceptor degeneration, geographic atrophy, and/or exudation with choroidal neovascularization in the macula. While several risk factors, including age, race, smoking, diet, oxidative stress and inflammation have been linked to AMD, the etiology and pathogenesis of the disease remain largely unclear. Current knowledge has shown that AMD development is strongly influenced by genetic factors. Treatment options for neovascular AMD include intravitreal injections of anti-vascular endothelial growth factor (VEGF); however, treatment options for geographic atrophy AMD are extremely limited. In FY2014, we reported several single nucleotide polymorphisms in the TIM3/SYN3, RAD51B, and FBN2 genes associated with AMD. We documented aberrant expression of interleukin-17A (IL17A) and the receptor IL17RC in the macula of AMD patients. In vitro, IL17A induces RPE cell death characterized by the accumulation of cytoplasmic lipids and autophagosomes with subsequent activation of pro-apoptotic Caspase-3 and Caspase-9. This pathology is reduced by siRNA knockdown of IL17RC. We investigate the link among macrophage, inflammasome, and IL-17 in AMD. We also found therapeutic effectiveness of adeno-associated virus vector encoding soluble IL-17 receptor, pimento epithelium-derived factor (PEDF), and platelet-derived growth factor-C (PDGF-C) on our double deficient Ccl2 and Cx3cr1 on Crb1rd8 background mice, an AMD model. Although the findings in the mouse model may not necessarily translate to findings in humans, they could still provide useful information for our understanding AMD pathogenesis and investigating potential therapeutic targets. 2. Ocular Lymphoma: Primary vitreoretinal lymphoma (PVRL), previously called primary intraocular lymphoma (PIOL), is a rare and fatal ocular malignancy. The diagnosis of PVRL is often challenging as it can masquerade as chronic uveitis. We published an article describing a method to differentiate PVRL from uveitis by using the level of miRNA-155 in the vitreous, which was significantly lower in PVRL. In human tissues, miRNA-155 regulates numerous transcriptional factors such as FOXP3, NFKB1, TGFB1, and AKT1. The interaction of microRNA-155 and those transcriptional factors plays an important role in innate immune responses and tumorigenesis. However, vitreal miRNA level does not offer an advantage to the ratio of IL-10 and IL-6 levels for the diagnosis of PVRL. We reviewed 853 patients at the NIH Uveitis clinic between 2004-2012 and found 21 (2.5%) with neoplastic masquerade syndrome. They were more likely to be older, non-African Americans who presented with unilateral, posterior uveitis. We also reported the difficulty in diagnosing PVRL, which often requires a high degree of clinical suspicion by physician and more than one invasive procedure (63% of the 27 PVRL patients at NIH) to make the diagnosis. We also reported an association between autofluorescent granularity patterns (hyperautofluorescent spots) with active PVRL. 3. New Pathology and Pathogenesis of Ocular Diseases: In collaboration with Drs. H. Nida Sen of NEI, Deborah Goldstein of Northwestern University, and Janet Davis of Bascom Palmer Institute, we edited a Special Issue on Gender and Uveitis. This special issue includes 10 articles and attempts to identify gender- and sex-based differences in infectious and noninfectious autoimmune uveitis, for example, multiple sclerosis in women, juvenile idiopathic arthritis in girls, and syphilitic uveitis in HIV-infected male patients. Clinical manifestations and courses may appear differently between genders in certain uveitides. We raised the relevance of HIF expression in von Hippel-Lindau (VHL) associated retinal hemangioblastoma. We provided unique and new molecular pathology in the eyes with sympathetic uveitis, ocular sarcoidosis, and viral retinitis.

我们识别并定位患者标本和动物组织中的炎症、退行性和恶性细胞及其产物和相关分子。我们主要通过常规病理学、免疫组织化学和分子病理学来分析这些细胞及其产物。尖端技术的应用，如显微切割结合分子技术（PCR、RT-PCR、基因分型等）使我们能够提供更准确的病理诊断（评估）并了解疾病的发病机制。这些技术和研究结果将指导我们为患者选择最有针对性的治疗方法。我们还通过各种动物模型研究不同眼部疾病的机制。使用动物模型，我们可以评估新型治疗剂对各种眼部疾病的疗效。 2014财年，我们在研究中完成了以下工作： 1. 年龄相关性黄斑变性（AMD）的分子病理学： AMD 是全球老年人不可逆转的严重中心视力丧失的主要原因。 预计到 2020 年，AMD 将影响 300 万 50 岁以上的美国人。AMD 的病理特征是软玻璃膜疣、视网膜色素上皮 (RPE) 和感光器变性、地图样萎缩和/或渗出的积累黄斑区有脉络膜新生血管。虽然年龄、种族、吸烟、饮食、氧化应激和炎症等多种危险因素与 AMD 相关，但该疾病的病因和发病机制仍不清楚。目前的知识表明，AMD 的发展受到遗传因素的强烈影响。新生血管性 AMD 的治疗选择包括玻璃体内注射抗血管内皮生长因子 (VEGF)；然而，AMD 地图样萎缩的治疗选择极其有限。 2014 财年，我们报道了与 AMD 相关的 TIM3/SYN3、RAD51B 和 FBN2 基因中的几个单核苷酸多态性。我们记录了 AMD 患者黄斑中白细胞介素 17A (IL17A) 和受体 IL17RC 的异常表达。在体外，IL17A 诱导 RPE 细胞死亡，其特征是细胞质脂质和自噬体的积累，随后激活促凋亡的 Caspase-3 和 Caspase-9。 IL17RC 的 siRNA 敲低可减少这种病理现象。我们研究了 AMD 中巨噬细胞、炎症小体和 IL-17 之间的联系。我们还发现编码可溶性 IL-17 受体、甘椒上皮衍生因子 (PEDF) 和血小板衍生生长因子-C (PDGF-C) 的腺相关病毒载体对 Crb1rd8 背景下的双缺陷 Ccl2 和 Cx3cr1 具有治疗效果小鼠，AMD 模型。尽管小鼠模型中的发现不一定能转化为人类的发现，但它们仍然可以为我们了解 AMD 发病机制和研究潜在的治疗靶点提供有用的信息。 2. 眼部淋巴瘤： 原发性玻璃体视网膜淋巴瘤（PVRL）以前称为原发性眼内淋巴瘤（PIOL），是一种罕见且致命的眼部恶性肿瘤。 PVRL 的诊断通常具有挑战性，因为它可能伪装成慢性葡萄膜炎。我们发表了一篇文章，描述了一种通过使用玻璃体中 miRNA-155 水平来区分 PVRL 和葡萄膜炎的方法，而 PVRL 中的 miRNA-155 水平明显较低。在人体组织中，miRNA-155 调节多种转录因子，例如 FOXP3、NFKB1、TGFB1 和 AKT1。 microRNA-155 与这些转录因子的相互作用在先天免疫反应和肿瘤发生中发挥着重要作用。然而，玻璃体 miRNA 水平在诊断 PVRL 时并不比 IL-10 和 IL-6 水平比值更有优势。 我们回顾了 2004 年至 2012 年间 NIH 葡萄膜炎诊所的 853 名患者，发现 21 名 (2.5%) 患有肿瘤伪装综合征。他们更有可能是患有单侧后葡萄膜炎的老年非非裔美国人。我们还报告了诊断 PVRL 的困难，这通常需要医生高度临床怀疑和不止一种侵入性手术（NIH 27 名 PVRL 患者中的 63%）才能做出诊断。我们还报道了自发荧光粒度模式（超自发荧光点）与活跃 PVRL 之间的关联。 3.眼部疾病的新病理学和发病机制： 与博士合作。 NEI 的 H. Nida Sen、西北大学的 Deborah Goldstein 和巴斯科姆帕尔默研究所的 Janet Davis，我们编辑了关于性别和葡萄膜炎的特刊。本期特刊包括 10 篇文章，试图确定传染性和非传染性自身免疫性葡萄膜炎中基于性别和性别的差异，例如女性多发性硬化症、女孩幼年特发性关节炎以及 HIV 感染男性患者的梅毒性葡萄膜炎。在某些葡萄膜炎中，不同性别的临床表现和病程可能有所不同。我们提出了 HIF 表达在 von Hippel-Lindau (VHL) 相关视网膜血管母细胞瘤中的相关性。我们提供了关于交感性葡萄膜炎、眼部结节病和病毒性视网膜炎的独特且新的分子病理学。