Carboxyethylpyrrole-Ethanolamine Phospholipids as AMD Biomarkers

 描述（由申请人提供）：视网膜相关性黄斑变性（AMD）是一种复杂的疾病，是老年人失明的主要原因。许多携带AMD风险基因型的个体从未发生这种疾病，并且只有一小部分被诊断患有AMD的人进展为具有严重视力丧失的晚期疾病。眼科医生无法确定哪些患者会进展为晚期AMD。我们的长期目标是开发用于评估晚期AMD风险和监测AMD治疗的预后技术。我们之前的生物标志物分析已经证实了血浆蛋白氧化修饰如羧乙基吡咯（CEP）、羧甲基赖氨酸（CML）和戊糖苷的AMD生物标志物潜力，并且蛋白质组学和基因组AMD生物标志物在一起使用时比单独使用更有效。我们提出了两个探索性的目的，测试的假设，CEP衍生物的乙醇胺磷脂（CEP-EPs）是上级AMD生物标志物相比，CEP蛋白质修饰。我们的初步研究表明，像CEP蛋白，CEP-EP诱导血管生成通过Toll样受体2。此外，我们还开发了将大量CEP-EP有效转化为单一衍生物（即CEP-乙醇胺（CEP-ETN））的方法，并建立了定量血浆中CEP-ETN的LC MS/MS方法。我们测量血浆CEP-ETN的初步研究表明，在AMD中CEP-EP比正常血浆升高得多，比CEP蛋白更高。在目的1中，我们将通过LC MS/MS验证AMD血浆中升高的CEP-ETN，并证明AMD生物标志物单独和与基因组AMD标志物和CEP-蛋白、CML和戊糖苷组合的效用。该结果将提高AMD生物标志物的区分准确性。在目标2中，我们将使用MALDI成像质谱法证实CEP-EP衍生物在AMD眼组织中升高，并为未来评估AMD治疗剂的动物研究建立可量化的终点。优秀的资源包括一个大型的、特征鲜明的血浆储存库、调查团队的经验和世界一流的视觉研究环境。总体影响将很大，因为有效的生物标志物将改变AMD患者的管理。

 DESCRIPTION (provided by applicant): Age-related macular degeneration (AMD) is a complex disease and the leading cause of blindness in the elderly. Many individuals carrying AMD risk genotypes never develop the disease and only a fraction of those diagnosed with AMD progress to advanced disease with severe visual loss. Ophthalmologists cannot determine which patients will progress to advanced AMD. Our long-term goal is the development of prognostic technology for assessing advanced AMD risk and for monitoring AMD therapeutics. Our previous biomarker analyses have confirmed the AMD biomarker potential of plasma protein oxidative modifications such as carboxyethylpyrrole (CEP), carboxymethyllysine (CML) and pentosidine and that proteomic and genomic AMD biomarkers are more effective when used together than alone. We propose two exploratory aims that test the hypothesis that CEP-derivatives of ethanolamine phospholipids (CEP-EPs) are superior AMD biomarkers compared to CEP-protein modifications. Our preliminary studies suggest that like CEP-protein, CEP-EPs induce angiogenesis through toll-like receptor 2. Furthermore we have developed methodology for the efficient conversion of a multitude of CEP-EPs to a single derivative, namely CEP-ethanolamine (CEP-ETN) and have established a LC MS/MS method for quantifying CEP-ETN in plasma. Our pilot study measuring plasma CEP-ETN indicates CEP-EPs are much more elevated in AMD than normal plasma, more so than CEP-protein. In aim 1, we will validate by LC MS/MS elevated CEP-ETN in AMD plasma and demonstrate AMD biomarker utility both alone and in combination with genomic AMD markers and CEP-protein, CML and pentosidine. The results will enhance the discriminatory accuracy of AMD biomarkers. In aim 2, we will confirm that CEP-EP derivatives are elevated in AMD ocular tissues using MALDI imaging mass spectrometry, and establish quantifiable endpoints for future animal studies evaluating AMD therapeutics. Outstanding resources include a large, well-characterized plasma repository, the experience of the investigative team and a world-class vision research environment. The overall impact will be high, as effective biomarkers will transform AMD patient management.