LRRK2 and Other Novel Exosome Proteins in Parkinson's Disease

描述（由申请人提供）：人们普遍认识到，在临床人群中检测与帕金森病（PD）相关的疾病进展和生化途径的措施不足，缺乏有效的生物标志物阻碍了并预计将继续阻碍有效的神经保护（疾病延迟或停止）治疗的成功临床试验。遗传学和病理学研究已经确定了几种与迟发性PD相关的蛋白质，这些蛋白质提供了对可能是疾病核心的途径的深入了解。然而，由于疾病易感组织的可用性有限，这些蛋白质难以在PD病例中进行分析， 研究通常局限于死后衍生的标本。我们最近观察到，LRRK 2和其他与PD遗传和病理相关的蛋白质在从人类尿液样品中分离的外泌体中是可检测的。使用多重质谱方法，我们可以定量来自临床尿液外泌体样本的935种蛋白质，其中140种蛋白质通过途径分析与神经变性有关。该提议寻求1）确定在源自PD患者与对照的尿外泌体-蛋白质组中是否存在与PD易感性和/或进展相关的生物标志物，和2）确定在用有效的LRRK 2激酶抑制剂舒尼替尼治疗的个体的尿外泌体中LRRK 2表达和/或磷酸化是否显著降低（一种多激酶抑制剂化合物），以建立用于未来LRRK 2抑制剂临床试验的靶向效应的测定。将在重复队列中验证潜在的感兴趣的生物标志物，并将在未来的多中心研究中进一步探索最有前途的线索。 公共卫生相关性：超过一百万美国人可能患有帕金森病，这是一种无法治愈的毁灭性神经退行性疾病。该提案将寻求新的标志物来帮助疾病诊断和跟踪疾病进展，这可能直接使帕金森病患者受益，并有助于为未来的临床试验提供动力，以发现新的治疗方法。

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. PUBLIC HEALTH RELEVANCE: Over one-million Americans may be affected with Parkinson's disease, a devastating neurodegenerative disorder for which there is no cure. This proposal will seek out new markers to aid in disease diagnosis and tracking of disease progression, which may directly benefit patients affected with Parkinson's disease and help power future clinical trials for the discovery of new therapeutics.