Molecular And Immunopathology Of Experimental And Clinical Ocular Diseases

我们在患者标本和动物样本中识别和局部定位眼部炎症、变性和肿瘤细胞及其产物，并通过常规病理学、免疫组织化学和分子病理学对其进行分析。 显微切割结合PCR、RT-PCR、RQ-PCR和RFLP等分子生物学技术的应用，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，并指导我们为患者选择合适的治疗方法。 我们从各种眼部疾病的动物模型中了解了大量关于不同眼部疾病的机制。 利用动物模型，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，并选择针对人类患者群体中特定疾病的上级疗法。 于2009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是一种使视网膜衰弱的疾病，其在临床上表现为中心视力丧失，在病理上表现为玻璃疣的积累、RPE变性、感光细胞萎缩，并且一些表现为脉络膜新血管形成（CNV）。 虽然一些危险因素，包括年龄，种族，吸烟和饮食与AMD有关，但该疾病的病因和发病机制仍不清楚。 这种情况的治疗选择同样有限。 AMD的人类和动物模型中的分子病理学发现包括CFH、CX 3CR 1和ARMS 2/HtrA 1以及与炎症有关的其他分子和细胞（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）。 我们还在4个病例对照样本中证实了HtrA 1启动子多态性、吸烟和年龄相关性黄斑变性之间的显著关联。 此外，我们证明了HtrA 1在AMD存档切片的黄斑病变中的表达增强。 我们检测到了C。pneumoniae DNA来自148名晚期AMD患者和162名对照的外周血，以及来自59名AMD和16名年龄匹配的非AMD存档载玻片的显微切割的黄斑细胞。 我们发现AMD和C.虽然CFH基因型可能不直接参与肺炎衣原体感染的发病机制，肺炎感染介导的AMD。 2.眼部淋巴瘤： 我们提出了新的命名：视网膜淋巴瘤原发性眼内淋巴瘤，因为它起源于视网膜。 我们回顾这种高度恶性肿瘤，通常是B细胞型，其与预后不良，因为频繁的中枢神经系统（CNS）的参与。 最近的证据表明，假定的起源细胞是一个晚胚中心或早期后胚中心B细胞。 我们已经证实了白细胞介素-10升高眼液中的B细胞视网膜淋巴瘤和眼B细胞淋巴瘤细胞的IgH基因重排的患者。 眼底荧光血管造影及OCT检查对视网膜淋巴瘤的诊断有重要价值。 绝大多数原发性眼内淋巴瘤是恶性B细胞，而眼内T细胞淋巴瘤是罕见的。 我们报告一个鼻腔自然杀伤T细胞淋巴瘤转移到视网膜的病例。 3.眼科疾病的新病理和发病机制： 我们报告了几个新的病理结果，包括：角膜缘干细胞缺乏症引起的全身化疗与羟基脲，铜绿假单胞菌DNA在脉络膜视网膜病变与慢性肉芽肿性疾病，炎症成分在脉络膜无色素，复发性肉芽肿性巩膜炎的韦格纳病人谁失败的抗肿瘤坏死因子治疗，和自身免疫性视网膜病变的系统性红斑狼疮。 这些新的病理学和分子病理学发现为上述系统性疾病的各种眼部并发症提供了见解。 4.各种眼部疾病的实验模型： 我们报道了富含EPA和DHA的饮食可以改善Ccl 2/Cx 3cr 1缺陷小鼠（我们的AMD小鼠模型）视网膜病变的进展。 这些小鼠中的发现与长链n-3脂肪酸降低AMD风险的流行病学研究一致。 这些结果进一步为使用ω-3脂肪酸及其生物活性衍生物预防和治疗AMD提供了科学依据。 与NEI的Caspi，Gery，Hooks和Nussenblatt博士以及NCI的Restifo博士合作，发现并发表了几种新的眼部炎症和黑色素瘤免疫治疗模型和机制。 这些活动在其年度报告中作了进一步说明。

We identify and topographically localize ocular inflammatory, degenerated, and tumor cells, and their products, in patient specimens and animal samples and analyze them 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 RFLP, allows us to provide more accurate pathological diagnosis (assessment) and pathogenesis of the disease, and guides us in selecting an appropriate therapy for the patient. We learn a great deal about the mechanisms of different ocular diseases from various animal models with ocular disorders. Using animal models, we can also access the efficacies of different therapeutic agents for various ocular diseases. This helps us better understand disease mechanisms and select superior therapies that target specific diseases in our human patient population. In FY2009, we accomplished the following in our research: 1. Molecular Pathology of Age-Related Macular Degeneration (AMD): AMD is a debilitating disease of the retina, which manifests clinically with the loss of central vision and pathologically with the accumulation of drusen, RPE degeneration, photoreceptor atrophy, and some with choroidal neovascularization (CNV). 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 the condition are similarly limited. The molecular pathological findings in both humans and animal models of AMD include genetic variations in CFH, CX3CR1, and ARMS2/HtrA1 and other molecules and cells implicated in inflammation (e.g., macrophages and microglia), apoptosis, cholesterol trafficking, angiogenesis (e.g., VEGF expression), and oxidative stress (e.g., the role of mitochondria). We also confirmed a significant association between the HtrA1 promotor polymorphism, smoking, and age-related macular degeneration in four case-control samples. Furthermore, we demonstrated enhanced expression of HtrA1 in the macular lesions from AMD archived slides. We detected C. pneumoniae DNA from the peripheral blood of 148 advanced AMD patients and 162 controls and from microdissected macular cells from 59 AMD and 16 age-matched, non-AMD archived slides. We found a possible association between AMD and C. pneumoniae infection, although the CFH genotype may not be directly involved in the pathogenesis of C. pneumoniae infection-mediated AMD. 2. Ocular Lymphoma: We have proposed new nomenclature: Retinal Lymphoma for primary intraocular lymphoma, as it originates in the retina. We review this high-grade malignancy, usually of B-cell type, and its association with a poor prognosis because of frequent central nervous system (CNS) involvement. Recent evidence suggests that the putative cell of origin is either a late-germinal center or an early post-germinal center B-cell. We have confirmed elevations of interleukin-10 in ocular fluids in patients with B-cell retinal lymphoma and the IgH gene rearrangements of ocular B-cell lymphoma cells. Ocular images (fluorescein angiogram and OCT) are helpful adjuncts for diagnosis of retinal lymphoma. The vast majority of primary intraocular lymphomas are malignant B-cells, while intraocular T-cell lymphomas are rare. We reported a case of metastatic nasal natural killer T-cell (NKT-cell) lymphoma to the retina. 3. New Pathology and Pathogenesis of Ocular Diseases: We reported several novel pathologic findings including: limbal stem cell deficiency arising from systemic chemotherapy with hydroxycarbamide, Pseudomonas aeruginosa DNA in a chorioretinal lesion associated with chronic granulomatous disease, an inflammatory component in choroideremia, recalcitrant granulomatous sclerouveitis in a Wegeners patient who failed anti-TNFα therapy, and an autoimmune retinopathy in systemic lupus erythematosus. These new pathological and molecular pathological findings provide insight to various ocular complications of the aforementioned systemic diseases. 4. Experimental Models for Various Ocular Diseases: We reported that a diet enriched in EPA and DHA can ameliorate the progression of retinal lesions in Ccl2/Cx3cr1 deficient mice, our mouse model of AMD. The findings in these mice are in line with the epidemiological studies of AMD risk reduction by long chain n-3 fatty acids. These results further provide the scientific basis for the use of omega-3 fatty acids and their biologically active derivatives in the prevention and treatment of AMD. In collaboration with Drs. Caspi, Gery, Hooks, and Nussenblatt of the NEI, and Dr. Restifo of the NCI, several new models and mechanisms of ocular inflammation and immunotherapy in melanoma have been discovered and published. These are further described in their annual reports.