Detection of Post-Translationally Modified Proteins as a Biomarker Panel for Parkinson's Disease

项目简介（由申请人提供）：项目主任/首席研究员（Walt, David， R）：项目摘要迄今为止，旨在确定帕金森病（PD）生物标志物的研究在很大程度上集中在测量血清或脑脊液中α -突触核蛋白或DJ-1等给定蛋白质的所有同种异构体的水平。然而，这些方法还不具备作为疾病指标所必需的敏感性和特异性。最近的证据表明，特定的翻译后修饰，如磷酸化和泛素化是PD病理生理的关键。这些翻译后修饰的蛋白尚未在血清中测量，因为这些同种异构体的浓度可能低于常规elisa的检测极限。我们假设检测翻译后修饰蛋白对PD生物标志物的开发至关重要。因此，我们将使用我们实验室新开发的技术，该技术比传统的ELISA检测这些异构体的灵敏度高100-1000倍。这项新技术，单分子阵列（SiMoA），是一种数字ELISA，可以对溶液中的单个分子进行计数。本课题旨在开发针对PD相关翻译后修饰蛋白的SiMoAs。我们将创建一种简单的血液测试，并使用PD患者、健康对照者和PDBP提供的其他运动障碍患者的血清样本评估其敏感性和特异性。总之，该应用程序寻求创建一个生物标志物面板，它是敏感的，特异性的，非侵入性的，并且比目前的诊断方法便宜得多。

DESCRIPTION (provided by applicant): Program Director/Principal Investigator (Walt, David, R): Project Abstract To date, research aimed at identifying a biomarker for Parkinson's Disease (PD) has, for the most part, focused on measuring the levels of all isoforms of a given protein such as alpha-synuclein or DJ-1 in serum or CSF. These methods, however, have not had the sensitivity and specificity necessary to be used as an indicator of disease. Recent evidence suggests that specific post-translational modifications such as phosphorylation and ubiquitination are key to the pathophysiology of PD. These post-translationally modified proteins have not yet been measured in serum because the concentrations of these isoforms are likely below the limit of detection of conventional ELISAs. We hypothesize that the detection of post-translationally modified proteins will be critical to PD biomarker development. Therefore, we will use technology, newly developed in our lab, which is 100-1000 times more sensitive than traditional ELISA to detect these isoforms. This new technology, Single Molecule Arrays (SiMoA), is a digital ELISA that counts individual molecules in solution. This proposal seeks to develop SiMoAs specific for post-translationally modified proteins relevant to PD. We will create a simple blood test and assess its sensitivity and specificity using serum samples from PD patients, healthy controls and individuals with other movement disorders provided by the PDBP. In sum, this application seeks to create a biomarker panel that is sensitive, specific, noninvasive and substantially cheaper than current diagnostic methods.