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

描述(由申请人提供)：项目主任/首席研究员(Walt，David，R)：项目摘要到目前为止，旨在确定帕金森氏病(PD)生物标记物的研究主要集中在测量特定蛋白质的所有异构体的水平，如血清或脑脊液中的α-突触核蛋白或DJ-1。然而，这些方法还没有作为疾病指示剂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，是帕金森病病理生理学的关键。这些翻译后修饰的蛋白质还没有在血清中被检测到，因为这些异构体的浓度很可能低于传统ELISA的检测极限。我们假设，翻译后修饰蛋白的检测将是PD生物标记物开发的关键。因此，我们将使用我们实验室新开发的技术来检测这些异构体，该技术的灵敏度比传统的ELISA高100-1000倍。这项名为单分子阵列(SiMoA)的新技术是一种数字酶联免疫吸附试验，可以对溶液中的单个分子进行计数。该提案寻求开发专用于与帕金森病相关的翻译后修饰蛋白的SiMoAs。我们将创建一种简单的血液测试，并使用PDBP提供的PD患者、健康对照组和有其他运动障碍的个人的血清样本来评估其敏感性和特异性。总之，这项申请寻求创建一种灵敏、特异、非侵入性的生物标志物面板，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.