Proteomic Trajectory Mapping of Retinopathy of Prematurity

描述（由申请人提供）：脊椎动物视网膜在出生后经历剧烈的转变，成为功能齐全的成年形式。在出生后的发育过程中，视网膜细胞增殖并在神经视网膜中形成网络。健康的发育对于成年期视网膜功能的充分表现至关重要。早产儿暴露于高氧，一旦恢复到正常氧水平（常氧），会引起视网膜病变。这种疾病被称为“早产儿视网膜病变”或ROP。ROP是由高氧条件消除后常氧诱导的视网膜新生血管形成引发的，并且随之发生的眼科并发症包括出血、新血管侵入视网膜外，最终发生视网膜脱离。氧诱导视网膜病变（OIR）的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的小鼠模型来理解视网膜病变的分子途径，并使用该模型来建立测定方案以评估将抑制ROP进展的药物。为此，我们使用了一种新的技术，称为“蛋白质组学轨迹映射”，已开发的PI的小组。蛋白质组学轨迹映射描述了每个视网膜蛋白质沿着发育时间轴的表达。我们假设ROP将表现在蛋白质组轨迹的改变中，某些药物将逆转改变过程。为了评估这一假设，我们将使用OIR的小鼠模型，并提出以下三个目标，在两年内。具体目标1：定义与正常视网膜发育相比OIR小鼠模型中的蛋白质组学轨迹。具体目标二：在蛋白质水平上通过蛋白质印迹和在mRNA水平上通过真实的-时间PCR来研究视网膜发育及其在OIR中的改变的基础的关键蛋白质和基因的表达。具体目标3：细胞定位的六个转录因子/调节，已确定我们的蛋白质组学实验将在正常和OIR视网膜免疫组化研究。这些目标的实现将为我们提供有关ROP发病机制的分子途径的重要信息。本研究结果将为ROP及其他相关视网膜疾病的治疗药物的开发做出重要贡献。

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.