Molecular And Immunopathology Of Experimental And Clinical Ocular Diseases

我们在患者标本和动物组织中识别和拓扑定位炎性、退行性和恶性细胞，以及它们的产物和相关分子。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）和了解疾病的发病机制。这些技术和发现将指导我们为患者选择最有针对性的治疗方法。我们还从各种动物模型中研究了不同眼部疾病的机制。使用动物模型，我们可以评估各种眼部疾病的新型治疗药物的疗效。于2014财政年度，我们在研究方面取得以下成果： 1.视网膜相关黄斑变性（AMD）的分子病理学： AMD是世界上老年人中不可逆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的病理学特征在于黄斑中软玻璃疣、视网膜色素上皮（RPE）和光感受器变性、地图状萎缩和/或渗出与脉络膜新血管形成的累积。虽然包括年龄、种族、吸烟、饮食、氧化应激和炎症在内的几个风险因素与AMD有关，但该疾病的病因和发病机制在很大程度上仍不清楚。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，地图状萎缩AMD的治疗选择非常有限。 在2014财年，我们报告了与AMD相关的TIM 3/SYN 3、RAD 51 B和FBN 2基因中的几种单核苷酸多态性。我们记录了白细胞介素-17A（IL 17 A）和受体IL 17 RC在AMD患者黄斑中的异常表达。在体外，IL 17 A诱导RPE细胞死亡，其特征在于细胞质脂质和自噬体的积累，随后激活促凋亡Caspase-3和Caspase-9。这种病理学通过IL 17 RC的siRNA敲低而减少。我们研究AMD中巨噬细胞、炎性小体和IL-17之间的联系。我们还发现了编码可溶性IL-17受体、甜椒上皮衍生因子（PEDF）和血小板衍生生长因子-C（PDGF-C）的腺相关病毒载体对我们的Crb 1 rd 8背景小鼠（AMD模型）上的Ccl 2和Cx 3cr 1双缺陷的治疗效果。虽然小鼠模型中的发现可能不一定转化为人类的发现，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），是一种罕见的和致命的眼部恶性肿瘤。PVRL的诊断通常具有挑战性，因为它可以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的miRNA-155水平来区分PVRL和葡萄膜炎的方法，该水平在PVRL中显著较低。在人类组织中，miRNA-155调节许多转录因子，如FOXP 3，NFKB 1，TGFB 1和AKT 1。microRNA-155与这些转录因子的相互作用在天然免疫应答和肿瘤发生中起着重要作用。然而，对于PVRL的诊断，玻璃体miRNA水平并不提供比IL-10和IL-6水平的比率更大的优势。 我们回顾了2004-2012年间NIH葡萄膜炎诊所的853例患者，发现21例（2.5%）患有肿瘤伪装综合征。他们更可能是年龄较大的非非洲裔美国人，表现为单侧后葡萄膜炎。我们还报告了诊断PVRL的困难，这通常需要医生高度的临床怀疑和一种以上的侵入性手术（NIH 27例PVRL患者中的63%）才能做出诊断。我们还报告了自发荧光粒度模式（超自发荧光点）与活动PVRL之间的关联。 3.眼科疾病的新病理和发病机制： 与Dr. H合作。NEI的Nida Sen，西北大学的Deborah Goldstein和巴斯科姆帕尔默研究所的Janet Davis，我们编辑了一个关于性别和葡萄膜炎的特刊。这期特刊包括10篇文章，并试图确定感染性和非感染性自身免疫性葡萄膜炎的性别和性别差异，例如，女性多发性硬化症，女孩青少年特发性关节炎，以及艾滋病毒感染男性患者的梅毒葡萄膜炎。在某些葡萄膜炎中，性别之间的临床表现和病程可能不同。我们提出了HIF表达在VHL相关视网膜血管母细胞瘤中的相关性。我们提供了独特的和新的分子病理学的眼睛与交感神经葡萄膜炎，眼结节病，病毒性视网膜炎。

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.