The treatment of uveitic cystoid macular edema with topical Interferon gamma

局部干扰素γ治疗葡萄膜炎性黄斑囊样水肿

• 批准号: 9550243
• 负责人: Sheldon Miller
• 金额: $ 9.98万
• 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/9550243
• 关键词: Age; Angiogenic Factor; Anterior; Bathing; Biological Assay; Blood capillaries; Cell Survival; Cells; Cornea; Cystoid Macular Edema; Disease; Drops; Edema; Endothelial Cells; Equilibrium; Eye; Eyedrops; Fluorescein; Hour; Human; Hydration status; Image; Immune system; In Vitro; Inflammasome; Inflammation; Inflammatory; Injectable; Interferon Type II; Interferons; Interleukin-18; Interleukin-6; Lead; Liquid substance; Measures; MicroRNAs; Optical Coherence Tomography; Pathogenesis; Pathologic; Pathway interactions; Patients; Phosphate Buffer; Physiologic Intraocular Pressure; Play; Resistance; Retinal; Retinal Detachment; Rodent Model; Role; Saline; Secondary to; Series; Side; Signal Transduction Pathway; Staining method; Stains; Structure of retinal pigment epithelium; Surface; Testing; Therapeutic Effect; Thick; Time; Tissues; Toxic effect; Uveitis; Visual Acuity; absorption; angiogenesis; body system; capillary; cell growth; conjunctiva; cytokine; experimental study; in vivo; irritation; macula; monolayer; ocular surface; primary outcome; receptor; response; secondary outcome

We hypothesize that proper retinal hydration is maintained by a balance of the bimodal functions of interferon gamma on the retinal pigment epithelium. Mounting evidence strongly suggests that the immune system plays an important role in angiogenesis. Pro-inflammatory cytokines, such as IFNg, IL-6, TNFa and IL-1b are the major cytokines in the pathogenesis of ocular inflammatory diseases and been shown to have receptors on RPE. TNFa, IL-6 and IL-1b are regarded as pro-angiogenic factors. However, IFNg is widely accepted as an anti-angiogenic cytokine due to its inhibitory effect on endothelial cell growth and capillary formation in other organ systems. We have evaluated the effect of interferon gamma on the JAK/STAT pathway, a signal transduction pathway also present in Human RPE cells. Fluid transport assays were performed to examine whether IFN induced changes in fluid transport across hfRPE monolayers. IFNg (10 ng/ml) addition to the basal bath increased JV by 8.6 ulcm-2hr-1, reflecting an increase in fluid absorption from the retinal to the choroidal side of the tissue. There were no apparent changes in cell viability as measured by transepithelial potential (TEP) and total tissue resistance (RT). In 10 experiments, the mean JV increased from 12.9 +/- 1.6 to 20.5 +/- 3.1 uL*cm-2*hr-1 (mean +/- s.e.m., P< 0.01). A previously tested in vivo rodent model of retinal detachment was used to measure the effect of INFg on re-absorption following retinal detachment. Initial detachment was created by injecting approximately 1ul of osmotically-balanced, modified phosphate-buffered saline (MPBS) solution into the sub-retinal space (SRS). Detachments that did not change in volume more than 10% in the first 30 minutes were used to test INFg effects. After detachment stabilization volume was measured by OCT imaging, INFg (40ul of 100ng/ml) was added to the anterior surface of the eye via eye drops (Celluvisc). A series of 3D OCT images were recorded at different time point. Addition of INFg to the anterior eye surface caused a significant, rapid 50% decrease in retinal detachment volume in the first hour of observation. This result is consistent with the observed fluid transport increase in RPE cells in vitro. We hypothesize that in the normal RPE cell, both pathways are functioning simultaneously to maintain the normal retinal hydration. The Jak/Stat pathway, if stimulated from basal side, will induce fluid absorption to resolve the abnormal accumulation of edema. Other regulatory targets of Jak/Stat pathway include micro RNA 155(miR-155), inflammasome, and IL-18 secretion. We also hypothesize that monotonic elevation of steady-state miR-155 levels in RPE occur with age, contributing to constitutive inflammasome activation. Modulation of IL-18 and IFNg levels may lead to therapeutic effect to reverse pathological changes associated with uveitis.

我们假设适当的视网膜水合是通过视网膜色素上皮上干扰素γ的双峰功能的平衡来维持的。越来越多的证据强烈表明免疫系统在血管生成中发挥着重要作用。促炎细胞因子，如 IFNg、IL-6、TNFa 和 IL-1b 是眼部炎症疾病发病机制中的主要细胞因子，并已被证明在 RPE 上有受体。 TNFa、IL-6和IL-1b被认为是促血管生成因子。然而，由于 IFNg 对其他器官系统的内皮细胞生长和毛细血管形成具有抑制作用，因此 IFNg 被广泛认为是一种抗血管生成细胞因子。我们评估了干扰素 γ 对 JAK/STAT 通路的影响，JAK/STAT 通路是人 RPE 细胞中也存在的信号转导通路。进行液体转运测定以检查 IFN 是否诱导跨 hfRPE 单层的液体转运变化。 基础浴中添加 IFNg (10 ng/ml) 可使 JV 增加 8.6 ulcm-2hr-1，反映从视网膜到组织脉络膜侧的液体吸收增加。通过跨上皮电位（TEP）和总组织电阻（RT）测量，细胞活力没有明显变化。在 10 个实验中，平均 JV 从 12.9 +/- 1.6 增加到 20.5 +/- 3.1 uL*cm-2*hr-1（平均值 +/- s.e.m.，P< 0.01）。 使用先前测试的视网膜脱离体内啮齿动物模型来测量INFg对视网膜脱离后重吸收的影响。通过将约 1ul 渗透平衡的改良磷酸盐缓冲盐水 (MPBS) 溶液注入视网膜下腔 (SRS) 来产生初始脱离。在前 30 分钟内体积变化不超过 10% 的分队用于测试 INFg 效果。 通过OCT成像测量脱离稳定体积后，通过滴眼剂（Celluvisc）将INFg（40ul，100ng/ml）添加到眼睛的前表面。在不同时间点记录一系列 3D OCT 图像。在观察的第一个小时内，将 INFg 添加到眼前表面导致视网膜脱离体积显着、快速减少 50%。这一结果与体外观察到的 RPE 细胞液体转运增加一致。 我们假设在正常的 RPE 细胞中，两条通路同时发挥作用以维持正常的视网膜水合作用。如果从基底侧刺激Jak/Stat通路，将诱导液体吸收以解决水肿的异常堆积。 Jak/Stat 通路的其他调控靶点包括微小 RNA 155(miR-155)、炎性体和 IL-18 分泌。我们还假设 RPE 中稳态 miR-155 水平随着年龄的增长而单调升高，从而导致组成性炎症小体激活。 IL-18 和 IFNg 水平的调节可能会产生逆转葡萄膜炎相关病理变化的治疗效果。