LRRK2 and Other Novel Exosome Proteins in Parkinson's Disease

描述(申请人提供)：人们普遍认为，在临床人群中检测与帕金森氏病(PD)相关的疾病进展和生化途径的措施不足，并且缺乏有效的生物标记物已经并预计将继续阻碍有效的神经保护(疾病延缓或停止)疗法的成功临床试验。遗传学和病理学研究已经确定了几种与晚发性帕金森病有关的蛋白质，这些蛋白质为深入了解可能是疾病中心的途径提供了洞察力。然而，由于疾病易感组织的有限，这些蛋白质很难在帕金森病病例中进行分析，并且 研究通常局限于死后提取的标本。我们最近观察到，LRRK2和其他在遗传和病理上与帕金森病有关的蛋白质，可以在从人类尿液样本中分离的外切体中检测到。利用多重质谱学方法，我们可以对临床尿液外切体样本中的935个蛋白质进行定量，通过通径分析，其中140个蛋白质与神经退行性变有关。这项建议旨在1)确定与PD易感性和/或进展相关的生物标志物是否来自PD患者和对照，以及2)确定LRRK2表达和/或磷酸化是否在接受有效的LRRK2激酶抑制剂Sunitinib(一种多激酶抑制剂化合物)治疗的患者的尿外蛋白体中显著降低，以建立一种靶向效应的检测方法，用于未来的LRRK2抑制剂临床试验。潜在的感兴趣的生物标志物将在复制队列中得到验证，最有希望的线索将在未来的多中心研究中进一步探索。

DESCRIPTION (provided by applicant): It is generally recognized that there are inadequate measures of detecting disease progression and biochemical pathways associated with Parkinson's disease (PD) in clinical populations, and that this lack of effective biomarkers has hindered, and is expected to continue to impede, successful clinical trials for efficacious neuroprotective (disease-retarding or halting) therapeutics. Genetic and pathological studies have identified several proteins linked to late-onset PD, and these proteins have provided insight into pathways that might be central to disease. However, these proteins have been difficult to analyze in PD cases due to the limited availability of disease-susceptible tissue, and studies are usually confined to post-mortem derived specimens. We have made the recent observation that LRRK2, and other proteins linked genetically and pathologically to PD, are detectable in exosomes isolated from human urine samples. Using a multiplex mass spectrometry approach, we can quantify 935 proteins from clinical urine exosome samples, and 140 of these proteins are linked to neurodegeneration by pathway analysis. This proposal seeks to 1) determine whether there are biomarkers associated with PD susceptibility and/or progression in urinary exosome-proteomes derived from PD patients versus controls, and 2) to determine if LRRK2 expression and/or phosphorylation are significantly lowered in the urinary exosomes of individuals treated with the potent LRRK2 kinase inhibitor sunitinib (a multi-kinase inhibitor compound), to establish an assay for on-target effects for future LRRK2 inhibitor clinica trials. Potential biomarkers of interest will be validated in replication cohorts, and the most promising leads will be further explored in future multi-center studies.