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: 8339745
• 负责人: Chi-Chao Chan
• 金额: $ 149.47万
• 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/8339745
• 关键词: Adult; Affect; Age; Age related macular degeneration; Animal Model; Annual Reports; Anti-Inflammatory Agents; Apoptosis; Area; B-Cell Lymphomas; Biological Models; Blindness; CX3CL1 gene; Candidate Disease Gene; Cells; Choroidal Neovascularization; Clinical; Collaborations; Copy Number Polymorphism; DNA Repair; Data; Development; Diagnosis; Diet; Disease; Drusen; Elderly; Elements; Endotoxins; Etiology; Experimental Models; Exudative age-related macular degeneration; Eye; Eye Neoplasms; Eye diseases; Genetic; Genotype; Goals; Granuloma; Hereditary Disease; Histopathology; Human; Human T-lymphotropic virus 1; Immunohistochemistry; Individual; Inflammation; Inflammatory; Injection of therapeutic agent; Interleukin-17; Intraocular Lymphoma; Invaded; Investigation; Knowledge; Learning; Lesion; Link; Liquid substance; Lymphoma; Mediating; Microdissection; Microglia; Mitochondria; Modeling; Molecular; Molecular Analysis; Mus; Naloxone; National Heart, Lung, and Blood Institute; National Human Genome Research Institute; National Institute of Allergy and Infectious Disease; Neoplasm Metastasis; Neoplasms; Nerve Degeneration; Nodule; Oxidative Stress; Pathogenesis; Pathology; Pathway interactions; Patients; Play; Publishing; Race; Reporting; Research; Research Personnel; Retina; Retinal; Retinal Degeneration; Retinal Lymphoma; Reverse Transcriptase Polymerase Chain Reaction; Risk Factors; Role; Safety; Series; Single Nucleotide Polymorphism; Smoking; Specimen; Sympathetic Ophthalmia; T-Cell Leukemia; T-Cell Lymphoma; Techniques; Technology; Therapeutic Agents; Thromboplastin; Tissues; United States; Uveitis; Vascular Endothelial Growth Factors; Von Hippel-Lindau Syndrome; Work; angiogenesis; animal tissue; autoimmune uveitis; cancer cell; chemokine; cholesterol trafficking; conventional therapy; cytokine; effective therapy; gene therapy; geographic atrophy; immunopathology; improved; leukemia/lymphoma; macrophage; molecular pathology; novel; novel therapeutics; photoreceptor degeneration; ultraviolet damage

We identify and topographically localize inflammatory, degenerative, and malignant cells, as well as their products in patient specimens and animal tissues. We analyze the 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 including PCR, RT-PCR, RQ-PCR and genotyping, allows us to provide more accurate pathological diagnosis (assessment) and pathogenesis of the disease. It also guides us in selecting the most appropriate treatment for patients. We learn a great deal about the mechanisms of different ocular diseases from various animal models. Using animal models, we can also assess the efficacy of different therapeutic agents for various ocular diseases. This helps us better understand disease mechanisms and select specific therapies targeting particular diseases for our patients. In FY2011, we accomplished the following in our research: 1. Molecular Pathology of Age-Related Macular Degeneration (AMD): AMD is the leading cause of vision loss among the elderly in the United States, and the number of individuals affected is expected to increase 50% by the year 2020. The pathology of AMD is characterized with the accumulation of drusen, RPE and photoreceptor degeneration, geographic atrophy, and/or exudation with choroidal neovascularization. While several risk factors, including age, race, smoking, and diet have been linked to AMD, the etiology and pathogenesis of the disease remain largely unclear. Treatment options for geographic atrophy AMD are extremely limited and for exudative/neovascular AMD include intravitreal injections of anti-vascular endothelial growth factor (VEGF). Current knowledge has revealed that AMD development is strongly influenced by genetic factors. We and other investigators have reported significant associations between AMD and single nucleotide polymorphisms in CFH, CX3CR1, and ARMS2/HtrA1 over the past few years. Growing evidence also suggests that inflammatory and immunological elements (e.g., macrophages and microglia), apoptosis, cholesterol trafficking, angiogenesis (e.g., VEGF expression), and oxidative stress (e.g., the role of mitochondria) play an important role in AMD development. In FY2011, using a duplex RT-PCR copy number genotyping approach, we evaluated copy number variations in 131 patients with exudative/neovascular AMD, but did not find definitive associations in 8 highly associated AMD candidate genes. We reported an increase of tissue factor expression in our murine model (Ccl2/Cx3cr1 double deficient mice) and human AMD lesions. We demonstrated that naloxone ameliorated retinal lesions of the Ccl2/Cx3cr1 double deficient mice via its inhibitory effect on microglia, and that AAV-5 mediated sFLT01 gene therapy arrested retinal lesions of these mice. In addition, we published a comprehensive review on the role of anti-inflammatory agents in AMD treatment. Conventional therapy that focuses solely on inhibiting angiogenesis may not be optimal because of the inflammatory involvement in AMD. Anti-inflammatory agents, which target specific inflammatory pathways and molecules, could be used as promising adjunct agents combined with anti-VEGF therapy for exudative neovascular AMD and potential therapies for geographic atrophy AMD. More studies are required to evaluate the efficacy and safety of the anti-inflammatory agents. 2. Ocular Lymphoma: We retrospectively reviewed a series of systemic metastatic retinal lymphoma cases at the NEI from 1991 to 2009. They had similar clinical presentations as primary vitreoretinal lymphoma. Systemic T-cell lymphomas that invade the retina and vitreous are more aggressive than systemic B-cell lymphoma. Molecular analysis is more useful than vitreous cytokine levels for the diagnosis of this metastasis. We reported ocular manifestations and pathology of adult T-cell leukemia/lymphoma associated with human T-cell lymphotropic virus type 1 (HTLV-1). 3. New Pathology and Pathogenesis of Ocular Diseases: We reported ophthalmic manifestations and histopathology including novel retinal degenerative findings of xeroderma pigmentosa. The new findings illustrate the role of DNA repair in protection of the eyes from UV damage and neurodegeneration of the retina. We conducted the first investigation of inflammatory cytokines and chemokines in 15 eyes with sympathetic ophthalmia (SO) and revealed predominant M1 macrophages in choroidal granulomas and Dalen-Fuchs nodules. High levels of Il-17 were expressed within granulomas and presence of Th1 in non-granulomatous inflammation. Targeting Th1, M1 and IL-17 may represent effective therapy for SO. 4. Experimental Models for Various Ocular Diseases: In collaboration with Drs. Caspi, Egwuagu, Hejtmancik, and Nussenblatt of the NEI, Drs. Robert Adelstein and Warren Leonard of NHLBI, Dr. Nicholas Restifo of NCI, Dr. Philip Murphy of NIAID, and Dr. Charles Venditti of NHGRI, different models and mechanisms of ocular inflammation and genetic diseases have been evaluated and published. The data is further described in the annual reports of these senior investigators.

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。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细胞及其产物。 显微切割等尖端技术的应用结合PCR、RT-PCR、RQ-PCR和基因分型等分子技术，使我们能够提供更准确的病理诊断（评估）和疾病的发病机制。 它还指导我们为患者选择最合适的治疗方法。 我们从各种动物模型中了解了大量关于不同眼部疾病的机制。 使用动物模型，我们还可以评估不同治疗药物对各种眼部疾病的疗效。 这有助于我们更好地了解疾病机制并为患者选择针对特定疾病的特定疗法。 2011财年，我们的研究取得了以下成果： 1. 年龄相关性黄斑变性（AMD）的分子病理学： AMD是美国老年人视力丧失的主要原因，预计到2020年受影响的人数将增加50%。AMD的病理特征是玻璃膜疣、RPE和感光器变性的积累，地图样萎缩和/或渗出并伴有脉络膜新生血管。 虽然年龄、种族、吸烟和饮食等多种危险因素与 AMD 相关，但该疾病的病因和发病机制仍不清楚。 地图样萎缩性 AMD 的治疗选择极其有限，而渗出性/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)。 目前的知识表明，AMD 的发展受到遗传因素的强烈影响。 我们和其他研究人员在过去几年中报告了 AMD 与 CFH、CX3CR1 和 ARMS2/HtrA1 的单核苷酸多态性之间的显着关联。 越来越多的证据还表明，炎症和免疫因素（例如巨噬细胞和小胶质细胞）、细胞凋亡、胆固醇运输、血管生成（例如 VEGF 表达）和氧化应激（例如线粒体的作用）在 AMD 发展中发挥着重要作用。 2011 财年，我们使用双重 RT-PCR 拷贝数基因分型方法评估了 131 名渗出性/新生血管性 AMD 患者的拷贝数变异，但没有发现 8 个高度相关的 AMD 候选基因之间的明确关联。 我们报道了小鼠模型（Ccl2/Cx3cr1 双缺陷小鼠）和人类 AMD 病变中组织因子表达的增加。 我们证明纳洛酮通过其对小胶质细胞的抑制作用改善了 Ccl2/Cx3cr1 双缺陷小鼠的视网膜病变，并且 AAV-5 介导的 sFLT01 基因治疗阻止了这些小鼠的视网膜病变。 此外，我们还发表了关于抗炎药在 AMD 治疗中的作用的全面综述。 由于 AMD 涉及炎症，仅注重抑制血管生成的传统疗法可能不是最佳选择。 针对特定炎症途径和分子的抗炎药物可作为有前景的辅助药物，与抗 VEGF 疗法联合治疗渗出性新生血管性 AMD，以及治疗地图样萎缩性 AMD 的潜在疗法。 需要更多的研究来评估抗炎药的功效和安全性。 2. 眼部淋巴瘤： 我们回顾性分析了1991年至2009年NEI收治的一系列全身性转移性视网膜淋巴瘤病例，其临床表现与原发性玻璃体视网膜淋巴瘤相似。侵入视网膜和玻璃体的系统性 T 细胞淋巴瘤比系统性 B 细胞淋巴瘤更具侵袭性。 对于诊断这种转移，分子分析比玻璃体细胞因子水平更有用。 我们报告了与人类 T 细胞嗜淋巴细胞病毒 1 型 (HTLV-1) 相关的成人 T 细胞白血病/淋巴瘤的眼部表现和病理学。 3.眼部疾病的新病理学和发病机制： 我们报告了眼科表现和组织病理学，包括色素性干皮病的新的视网膜退行性发现。 新发现说明了 DNA 修复在保护眼睛免受紫外线损伤和视网膜神经变性方面的作用。 我们对 15 只患有交感性眼炎 (SO) 的眼睛进行了炎症细胞因子和趋化因子的首次研究，发现脉络膜肉芽肿和 Dalen-Fuchs 结节中主要存在 M1 巨噬细胞。 高水平的 Il-17 在肉芽肿内表达，并且在非肉芽肿性炎症中存在 Th1。 针对 Th1、M1 和 IL-17 可能是 SO 的有效治疗方法。 4.各种眼部疾病的实验模型： 与博士合作。 NEI 的 Caspi、Egwuagu、Hejtmancik 和 Nussenblatt，博士。 NHLBI的Robert Adelstein和Warren Leonard、NCI的Nicholas Restifo博士、NIAID的Philip Murphy博士和NHGRI的Charles Venditti博士对眼部炎症和遗传性疾病的不同模型和机制进行了评估并发表。 这些数据在这些高级研究人员的年度报告中得到了进一步描述。