Establishing clinical utility of CSF biomarkers for PD

项目摘要-项目3：建立脑脊液生物标记物对帕金森病的临床应用 帕金森病(PD)的准确诊断和对PD患者的监测仍然具有挑战性 排除了最有效的病人护理。尽管NINDS和其他PD患者倡导团体，如 迈克尔·J·福克斯基金会已经认识到诊断和预测帕金森病的生物标记物的必要性，没有 到目前为止，这种生物标记物已经得到了验证。这项临床研究项目将利用多种反应 监测-质谱学(MRM-MS)以确定是否可以使用特定的蛋白质磷酸化事件 作为新的帕金森病特异性诊断/预后生物标志物。 先前的研究表明，非受体酪氨酸激酶c-Abl使α-突触核蛋白磷酸化 在Y-39地点，在Y-143地点帕金。在帕金森病期间，这两条通路的失调最终导致 导致神经细胞死亡，进而导致帕金森病的临床表现。在初步研究中，我们有 已经开发了基于MRM-MS的定量分析来监测这些磷酸化和 脑脊液样本中α-突触核蛋白和帕金蛋白的非磷酸化形式。我们的假设是c-的调节失调 ABL信号级联与帕金森病的发病有内在联系，其相对的磷酸化状态 下游的c-abl底物，α-突触核蛋白和parkin，应该为存在提供一个灵敏的读数。 和/或帕金森病的严重程度。 目的1将测定α-突触核蛋白的Y-39胰蛋白酶多肽在其 帕金森病患者和对照组脑脊液中的磷酸化和非磷酸化形式。这个化验结果将是 在几个临床队列中用于确定磷酸化的Y-39残基是否可以起到(I)的作用 区分PD患者和对照组的诊断标记物和/或(Ii)PD预后标记物 患者可以用来评估PD的早期和晚期(更严重)。 目标2将采用目标1中描述的相同方法来测试c-abl介导的磷酸化 可用于辅助诊断帕金森氏病或作为以下功能的患者监测 疾病的严重性。 综上所述，该项目将确定和验证c-abl底物作为新的诊断/预后指标。 基于蛋白质的生物标记物有可能为临床医生提供新的策略来建立 明确诊断帕金森病和/或监测帕金森病患者的病情严重程度。

PROJECT SUMMARY- PROJECT 3: ESTABLISHING CLINICAL UTILITY OF CSF BIOMARKERS FOR PD Accurate diagnosis of Parkinson's disease (PD) and monitoring of PD patients remain challenging and preclude the most effective patient care. Although the NINDS and other PD patient advocacy groups such as the Michael J. Fox Foundation have recognized the need for both diagnostic and prognostic PD biomarkers, no such biomarkers have been validated thus far. This Clinical Research Project will utilize Multiple Reaction Monitoring-Mass Spectrometry (MRM-MS) to determine if specific protein phosphorylation events can be used as novel PD-specific diagnostic/prognostic biomarkers. Previous research has shown that the non-receptor tyrosine kinase, c-Abl, phosphorylates α–Synuclein at the Y-39 site and Parkin at the Y-143 site. During PD, dysregulation of these two pathways ultimately leads to neuronal cell death, which in turn leads to the clinical manifestations of PD. In preliminary studies, we have already developed MRM-MS based quantitative assays to monitor these phosphorylated and unphosphorylated forms of α-Synuclein and Parkin in CSF samples. Our hypothesis is that dysregulation of c- Abl signaling cascade is intrinsically linked to PD pathogenesis and that the relative phosphorylation state of downstream c-Abl substrates, α-Synuclein and Parkin, should provide a sensitive read-out for the presence and/or the severity of PD. Aim 1 will measure the relative concentration of the Y-39 tryptic peptide of α-Synuclein in both its phosphorylated and unphosphorylated forms in the CSF of PD patients and controls. This assay will be employed in several clinical cohorts to determine whether the phosphorylated Y-39 residue can function as (i) a diagnostic marker by differentiating between PD patients and controls and/or (ii) a prognostic PD marker in patients that can be used for assessment of early and late (more severe) stages of PD. Aim 2 will adopt the same approach as described in Aim 1 to test whether c-Abl mediated phosphorylation of Parkin at Y-143 can be used to facilitate either diagnosis with PD or patient monitoring as a function of disease severity. Taken together, this project will identify and validate c-Abl substrates as novel diagnostic/prognostic protein-based biomarkers that have the potential to provide clinicians with novel strategies for establishing a definitive diagnosis of PD and/or for monitoring the severity of disease in patients with PD.