Proteomic Trajectory Mapping of Retinopathy of Prematurity

描述(申请人提供)：脊椎动物的视网膜在出生后经历了剧烈的转变，变成了功能齐全的成体。在出生后的发育过程中，视网膜细胞的增殖和神经视网膜网络的形成发生了。健康的发育对于成人期视网膜功能的充分表现至关重要。早产儿暴露在高氧环境中，当恢复到正常的氧气水平(常氧)时，会导致视网膜病变。这种疾病被称为“早产儿视网膜病变”或ROP。ROP是在去除高氧状态后，由常氧诱导的视网膜新生血管触发的，继而发生的眼科并发症包括出血、新生血管侵入视网膜外，最终导致视网膜脱离。氧诱导视网膜病变(OIR)啮齿动物模型被广泛用作人类ROP的实验动物模型。我们的长期目标是使用OIR的小鼠模型来了解视网膜病变背后的分子途径，并利用该模型建立一种检测方案来评估将抑制ROP进展的药物。为此，我们使用了一种名为“蛋白质轨迹图”的新技术，这项技术是由PI的团队开发的。蛋白质组轨迹图描述了每种视网膜蛋白在发育时间轴上的表达。我们假设ROP将表现为蛋白质组轨迹的改变，某些药物将逆转这一改变过程。为了评估这一假设，我们将使用OIR的小鼠模型，并在两年内提出以下三个目标。具体目标1：确定OIR小鼠模型中的蛋白质组学轨迹，并与正常视网膜发育进行比较。具体目的2：通过蛋白质印迹和实时荧光定量聚合酶链式反应在蛋白质水平和实时荧光定量聚合酶链式反应的方法研究视网膜发育的关键蛋白和基因的表达及其在OIR中的变化。具体目标3：我们的蛋白质组学实验已经确定的六个转录因子/调节因子的细胞定位将通过免疫组织化学在正常和OIR视网膜中进行研究。这些目标的实现将为我们提供有关ROP发病机制的重要信息。这一结果将对开发治疗ROP和其他相关视网膜疾病的药物做出重大贡献。

DESCRIPTION (provided by applicant): The vertebrate retina undergoes drastic transformation after birth into the fully functional adult form. During postnatal development the proliferation of retinal cells and network formation in the neural retina take place. Healthy development is crucial to the full manifestation of retinal function in the adult stage. Exposure of premature infants to hyperoxia, upon returning to a normal oxygen level (normoxia), causes retinopathy. This disease is called "retinopathy of prematurity" or ROP. ROP is triggered by the retinal neovascularization induced by the normoxia after the hyperoxic condition is removed, and the consequent ophthalmologic complications takes place including bleeding, invasion of new vessels to outside of the retina, and finally retinal detachment. The rodent model of Oxygen Induced Retinopathy (OIR) is widely utilized as an experimental animal model for human ROP. Our long term goal is to understand the molecular pathways underlying the retinopathy using the mouse model of OIR and to use the model to establish an assay protocol to evaluate drugs that would suppress the progress of ROP. For this purpose we use a novel technology called "Proteomic Trajectory Mapping" that has been developed by the PI's group. Proteomic trajectory mapping describes the expression of each retinal protein along the developmental time axis. We hypothesize that ROP will be manifested in the alterations of the proteomic trajectories and certain drugs will reverse the alteration process. To evaluate this hypothesis, we will use the mouse model of OIR and propose the following three aims in the two year period. Specific Aim 1: To define proteomics trajectories in the mouse model of OIR in comparison to normal retinal development. Specific Aim 2: Expression of the key proteins and genes underlying the retinal development and its alteration in OIR will be investigated both at the protein level by western blot and at the mRNA level by real- time PCR. Specific Aim 3: Cellular localization of six transcription factors/regulators which have been identified by our proteomics experiments will be investigated by immunohistochemistry in both normal and OIR retinas. Achievement of these aims will give us crucial information on the molecular pathways that are involved in the pathogenesis of ROP. The results will contribute significantly to the development of drugs for the treatment of ROP and other related retinal diseases.