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

检测翻译后修饰蛋白作为帕金森病的生物标志物组

• 批准号: 9607157
• 负责人: DAVID R. WALT
• 金额: $ 28.61万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-15 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9607157
• 关键词: Detection; Post; Translationally; Modified; Proteins

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.

描述（由申请人提供）：计划主管/首席研究员（Walt，David，R）：迄今为止的项目摘要，旨在识别帕金森氏病的生物标志物（PD）在大多数情况下，重点是测量给定蛋白质的所有同工型，例如Alpha-synuclein或dj-1 in in Alpha-synnuclein或dj-1。但是，这些方法没有用作疾病指标所必需的敏感性和特异性。最近的证据表明，诸如磷酸化和泛素化等特定的翻译后修饰是PD病理生理学的关键。这些翻译后修饰的蛋白质尚未在血清中测量，因为这些同工型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的蛋白质对PD生物标志物的发育至关重要。因此，我们将使用在实验室中新开发的技术，该技术比传统ELISA敏感100-1000倍，以检测这些同工型。这项新技术，即单分子阵列（SIMOA），是一种数字ELISA，可计算溶液中的单个分子。该提案旨在开发针对与PD相关的翻译后修改后修饰的蛋白质的SIMOA。我们将使用来自PD患者的血清样本，健康对照组以及PDBP提供的其他运动障碍的个体来创建简单的血液测试，并评估其敏感性和特异性。总而言之，该应用程序旨在创建一个生物标志物面板，该面板比当前的诊断方法敏感，特异性，无创和便宜。