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定位炎性细胞、退行性细胞和恶性细胞及其产物和相关分子。我们主要通过常规病理、超微结构、免疫组织化学和分子病理学对这些细胞及其产物进行分析。尖端技术的应用，如显微解剖与分子技术(聚合酶链式反应、逆转录聚合酶链式反应、基因分型等)相结合。使我们能够提供更准确的病理诊断(评估)，并更好地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。我们还从不同的动物模型研究了不同眼病的发病机制。利用动物模型，我们可以评估新的治疗药物对各种眼病的疗效。2015财年，我们在研究方面完成了以下工作： 1.老年性黄斑变性的分子病理学： AMD是世界上导致老年人严重中心视力丧失的主要原因。到2050年，美国晚期AMD的估计人数预计将从207万人(截至2010年)增加到540万人。AMD的病理特征是黄斑软性玻璃体积聚、视网膜色素上皮(RPE)和感光细胞变性、地理性萎缩和/或渗出伴脉络膜新生血管。虽然包括年龄、种族、吸烟、饮食、氧化应激和炎症在内的几个危险因素与AMD有关，但这种疾病的病因和发病机制在很大程度上仍不清楚。目前的研究表明，AMD的发育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，对于地理性萎缩AMD的治疗选择极其有限。 在2015财年，我们报告了RAD51B、FPR1和UBE3D中新的单核苷酸多态与AMD的关联(参见Zia EY000418-12)。我们展示了在炎症和/或氧化应激下RPE细胞中的自噬小体形成、线粒体损伤、核外DNA的存在、细胞质孔的形成和NLRP炎性小体的移位，以及诊断为AMD的人眼。我们认为AMD的发病机制之一可能是由炎性细胞因子(炎症体IL1和IL18 IL17)和潜在的caspase-1介导的下垂/坏死性下垂介导的先天免疫激活，导致视网膜组织损伤和细胞死亡。 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主要起源于B细胞，其中的细胞渗透到眼睛和大脑。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。我们发表并综述了眼内淋巴瘤模型。动物模型有助于研究淋巴瘤细胞的生长，因为恶性淋巴细胞在体外几乎没有增殖能力。动物模型被用来显示对新型治疗药物的疗效，如抗B细胞单抗。我们还报告了一例罕见的PVRL合并肾脏、盆腔和中枢神经系统损害。 3.眼科疾病的新病理学和发病机制： 我们继续研究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。以男性为主的自身免疫性疾病，如Behets病，通常在50岁之前出现临床症状，以急性炎症和Th1反应为特征。人类白细胞抗原B27相关的男性前葡萄膜炎就是一个例子。女性为主的自身免疫，如在类风湿性关节炎和系统性红斑狼疮中发现的，倾向于在晚年出现，可能通过Th2和自身抗体相关机制发生。认识到自身免疫中的性别差异，包括葡萄膜炎，应该能够在未来为患者制定新的预防和治疗策略。 我们还展示了急性视网膜坏死眼以及von Hippel-Lindau病合并虹膜和角膜新生血管、肉芽肿和多血管炎的独特和新的分子病理学。 4.眼科疾病实验动物模型： 作为编辑，我们邀请了10位知名的视觉科学家和眼科医生共同撰写了一本书，书中描述了模拟常见人类眼部疾病的实验动物模型：疱疹病毒性角膜炎、白内障、青光眼、老年性黄斑变性、糖尿病视网膜病变、葡萄膜炎、视网膜色素变性、Graves病和眼内肿瘤。同时，这些模型反映了用于研究人类疾病的工具的多样性和实用性。世界专家临床医生根据他们的临床经验讨论每种模式。在描述眼科疾病最相关的动物模型时，这本书将引起眼科医生、视力研究人员、研究员、住院医生和医科学生的兴趣。这本书将于2015年出版。 我们在建立视网膜血管母细胞瘤转基因VHL小鼠模型方面取得了进展。我们使用基因打靶技术和cre-lox位点特异性重组系统来产生这个小鼠模型。四年前，我们开始了在血管母细胞启动子(SCL-CRE-ER(T))的控制下，将cre-重组酶基因与雌激素受体融合的小鼠，并与一只带有VHL基因两侧有loxP位点的小鼠杂交。然后，我们创造了一个小鼠，当我们在E9.5后(在P16天)给他莫昔芬时，cre-重组酶可以识别loxP位点，并有效地删除VHL基因。因此，我们可以在VHL胚胎致死的发育阶段诱导VHL的缺失，血管母细胞启动子将赋予我们细胞特异性。 随着陈博士的退休，免疫病理科于2015年5月关闭。

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, ultrastructure, 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 a better understanding of 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 FY2015, 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. By 2050, the estimated number of people with late AMD is expected to increase from 2.07 million (as of 2010) to 5.4 million in the USA. 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 indicates that AMD development is strongly influenced by genetic factors. Treatment options exist for neovascular AMD include intravitreal injections of anti-vascular endothelial growth factor (VEGF); however, treatment options for geographic atrophy AMD are extremely limited. In FY2015, we reported the associations of new single nucleotide polymorphisms in RAD51B, FPR1 and UBE3D and AMD (see ZIA EY000418-12). We illustrated autophagosome formation, mitochondrial damage, the presence of extranuclear DNA, cytoplasmic pore formation, and NLRP inflammasome translocation in the RPE cells under inflammatory and/or oxidative stress, as well as in human eyes diagnosed with AMD. We suggest that one of the sources of AMD pathogenesis can be the activation of innate immunity resulting in damage to retinal tissue and cell death mediated by inflammatory cytokines (inflammasome IL1β/IL18 IL17) and potentially caspase-1 mediated pyroptosis/necroptosis. 2. Ocular Lymphoma: Primary vitreoretinal lymphoma (PVRL), previously called primary intraocular lymphoma (PIOL), is a rare and fatal ocular malignancy. PVRL is mainly of B-cell origin wherein the cells infiltrate the eye and brain. The diagnosis of PVRL is often challenging as it can masquerade as chronic uveitis. We published and reviewed intraocular lymphoma models. Animal models aid in the study of lymphoma cell growth, as the malignant lymphocytes show little proliferative capacity in vitro. The animal models were used to show the efficacy to novel therapeutic agents, such as anti-B cell monoclonal antibodies. We also reported an unusual case of PVRL with renal, pelvic, and CNS lesions. 3. New Pathology and Pathogenesis of Ocular Diseases: We continue to study gender- and sex-based differences in infectious and noninfectious autoimmune uveitis. Male-predominant autoimmune diseases, such as Behets disease usually manifest clinically before the age of 50 and are characterized by acute inflammation and a Th1 response. HLA-B27 associated anterior uveitis in men is an example of this. Female-predominant autoimmunity, as found in rheumatoid arthritis and systemic lupus erythematosus tends to present later in life and likely occurs via Th2 and autoantibody related mechanisms. Recognition of the gender differences in autoimmunity, including uveitis, should allow new preventative and therapeutic strategies for patients in the future. We also illustrated unique and new molecular pathology in eyes with acute retinal necrosis as well as in von Hippel-Lindau disease with neovascularization of the iris and cornea, granulomatosis, with polyangitis. 4. Experimental animal models of ophthalmic diseases: As an editor, we invited 10 renown, visual scientists and ophthalmologist to co-author a book, which describes experimental animal models that mimic common human ocular diseases: herpetic keratitis, cataract, glaucoma, age-related macular degeneration, diabetic retinopathy, uveitis, retinitis pigmentosa, Graves disease, and intraocular tumors. In conjunction, these models reflect the diversity and utility of tools used to study human disease. World expert clinicians discuss each model based on their clinical experience. In describing the most pertinent animal models of ophthalmic diseases, this book will be of interest to ophthalmologists, vision researchers, fellows, residents, and medical students. This book will be published in 2015. We have shown progress in generation of a transgenic VHL mouse model with retinal hemangioblastoma. We used gene targeting technology and the cre-lox site-specific recombination system to generate this mouse model. Four years ago, we started out with mice with cre-recombinase gene fused with estrogen receptor under the control of an angioblast promotor (the SCL-Cre-ER(T)) and crossed with a mouse that has the VHL gene flanked by loxP sites. Then, we created a mouse in which when we give tamoxifen after E9.5 (at P16 day), where cre-recombinase can recognize the loxP sites and effectively delete the VHL gene. So here, we can induce the deletion of VHL past the time in development when it is embryonically lethal AND the angioblast promotor will give us cell specificity. The Immunopathology Section closed May 2015 with the retirement of Dr. Chan.