IL-10, Inflammation, and Selective INhibition of Neovascularization

描述（由申请人提供）：在糖尿病性视网膜病变和早产儿视网膜病变（ROP）中，视网膜血管损伤会产生视网膜缺氧区域，这些区域表达升高水平的促血管生成因子。奇怪的是，血管生成反应不仅诱导受损视网膜的再血管化，而且诱导病理性新生血管化（NV）进入玻璃体，在那里它可以导致失明。理想的治疗方法是抑制病理性NV而不损害视网膜内修复。由于VEGF是视网膜内和玻璃体内血管生长所必需的，因此目前靶向VEGF的疗法不能选择性抑制NV。我们最近发现，抗炎白细胞介素10（IL-10）是一种强大的和选择性的病理性NV抑制剂。在ROP的小鼠模型中，单次注射IL-10使NV降低80%，而不损害受损视网膜的血管再生。虽然IL-10具有短的半衰期（T1/2，在血浆中3小时），但它减少NV达5天。我们假设IL-10控制一个促炎过程，启动一个级联反应，引导血管生成远离视网膜并进入玻璃体。在肿瘤坏死因子α（TNF 1）受体、诱导型一氧化氮合酶（iNOS）或环氧合酶-2（考克斯-2）遗传缺陷的小鼠中已经证明了病理性NV的选择性抑制。IL-10在体外抑制这些在巨噬细胞中的表达。该研究将测试IL-10的选择性是否是剂量依赖性的，是否所有NV都可以在更高浓度下被阻止，以及它是否抑制视网膜中TNF 1、iNOS、考克斯-2或巨噬细胞趋化肽（MCP-1）的表达。目的1还将鉴定参与IL-10调节的血管生成的特定细胞类型（例如星形胶质细胞、小胶质细胞、巨噬细胞）。目的2将测试是否减少巨噬细胞募集到视网膜选择性抑制病理性NV。目的3将使用定量RT-PCR来阐明IL-10对促炎因子和促血管生成因子的视网膜表达的影响，使用已知的IL-10在巨噬细胞中的作用作为指导。 本研究的长期目标是使用ROP范式来识别驱动病理性NV的IL-10敏感事件，并最终找到治疗糖尿病视网膜病变和ROP的新疗法。

DESCRIPTION (provided by applicant): In diabetic retinopathy and retinopathy of prematurity (ROP), damage to the retinal blood vessels creates regions of retinal hypoxia that express elevated levels of proangiogenic factors. Paradoxically, the angiogenic response induces not only revascularization of the damaged retina, but also pathological neovascularization (NV) into the vitreous, where it can cause blindness. The ideal therapy would suppress pathological NV without damaging intraretinal repair. Since VEGF is required for both intraretinal and intravitreal vessel growth, current therapies targeting VEGF cannot selectively inhibit NV. We recently found that the anti-inflammatory interleukin 10 (IL-10) is a powerful and selective inhibitor of pathological NV. In a mouse model of ROP, a single injection of IL-10 reduced NV 80% without impairing revascularization of the damaged retina. Although IL-10 has a short half-life (T1/2, 3 hrs in plasma), it reduced NV for 5 days. We hypothesize that IL-10 controls a pro-inflammatory process that initiates a cascade that directs angiogenesis away from the retina and into the vitreous. Selective inhibition of pathological NV has been demonstrated in mice genetically deficient in tumor necrosis factor alpha (TNF1) receptors, inducible nitric oxide synthase (iNOS) or cyclooxygenase-2 (COX-2). IL-10 suppresses expression of these in macrophages in vitro. The proposed research will test if the selectivity of IL-10 is dose-dependent and whether all NV could be arrested at higher concentrations, and whether it suppresses TNF1, iNOS, COX-2 or macrophage chemotactic peptide (MCP-1) expression in the retina. Aim 1 will also identify specific cell types (e.g. astrocytes, microglia, macrophages) involved in IL-10 regulated angiogenesis. Aim 2 will test whether reduced macrophage recruitment into the retina selectively inhibits pathological NV. Aim 3 will use quantitative RT- PCR to elucidate the effect of IL-10 on retinal expression of pro-inflammatory and pro-angiogenic factors, using known effects of IL-10 in macrophages as a guide. The long range goal of this research is to use the ROP paradigm to identify IL-10 sensitive events that drive pathological NV, and ultimately to find new therapies for diabetic retinopathy and ROP.