The UCLA Udall Parkinson Disease Center of Excellence

描述(由申请人提供)： 新证据表明，不同的突变导致家族性帕金森病(PD)的机制也可能在散发性帕金森病(PD)中起作用。这些新数据表明细胞功能障碍在细胞死亡之前的重要性，以及非多巴胺能神经元在疾病中的参与。因此，识别多种原因共同的细胞功能障碍机制可能为阻止或逆转疾病进程提供新的治疗靶点。加州大学洛杉矶分校Udall Parkinson疾病卓越中心的这份续签申请侧重于在表达PD导致突变的补充模型中对功能障碍进展的研究，以及在具有良好特征的患者群体中的研究。该中心由5个项目组成，由一个行政核心和一个小鼠遗传学核心提供支持。在前三个项目中，我们建议继续在当前奖项的支持下进行协调的多学科工作，以表征帕金森病遗传小鼠模型中运动和非运动行为异常和神经病理学的进展(项目1)、神经递质释放异常(项目2)和突触功能(项目3)，包括基于BAG技术的新模型。这些项目将通过增加细胞模型(项目4)来补充，以分析导致在小鼠中观察到的表型的细胞功能障碍的机制。研究功能障碍的进展也将是该中心新的以患者为导向的部分的重点。在这个项目(项目5)中，我们将对诊断后的疾病表型进行临床纵向研究，包括精神和认知并存。这将与开发和验证经改进的与健康有关的生活质量评估工具相结合。这些以患者为中心的研究将为未来对来自相同患者的遗传物质的分析提供关键的临床数据，并将我们的基础研究努力转化为更好的患者护理。为了确定导致帕金森病神经变性的细胞变化，加州大学洛杉矶分校Udall Parkinson疾病卓越中心将专注于在运动症状出现之前疾病的早期表现及其进展。结合实验模型和临床研究，我们中心的目标是了解这些细胞功能障碍的机制，以刺激能够阻止疾病进程的治疗策略的发展。 项目1 标题：DA细胞死亡前行为和病理缺陷的进展 帕金森病小鼠模型的建立 PI：玛丽-弗朗索瓦·切塞特，医学博士，博士 描述(由申请人提供)： 新证据表明，不同的突变导致家族性帕金森病(PD)的机制也可能在散发性帕金森病(PD)中起作用。在目前的资助期间，我们已经在表达各种已知导致人类帕金森病的突变的小鼠中发现了没有细胞死亡的细胞功能障碍。在这一新的应用中，我们将测试这一假说，即常见的功能障碍机制可能是由不同的突变引起的，并在疾病的早期阶段提供治疗靶点，在进一步的细胞死亡导致不可逆转的损害之前。项目1.2和3构成了当前奖项的延续，并将继续使用多学科方法来揭示现有和新的小鼠模型中神经元功能障碍的机制。在项目1的特定目标1中，我们将确定已有的在不同启动子下过表达α-突触核蛋白的小鼠和PARKIN KO小鼠的行为缺陷和神经病理学的进展。我们将使用我们开发的一系列敏感的运动测试来评估小鼠的运动表型，并将这种分析扩展到非运动行为，因为相关症状可能会对患者的生活质量产生重大影响，如中心项目5所述。我们将使用免疫组织化学来确定阿尔法-突触核蛋白病理的进展和已知的帕金森病影响的多个脑区的神经胶质激活，并检测参与神经递质释放的蛋白的异常表达，并在中心的项目4中进行检测。我们将使用配体结合和分子方法来识别多巴胺能传递的失调，这将在项目2中用神经化学方法进一步检查，在项目3中用电生理学进一步检查。最后，将在无偏见体视学的老年动物中评估受帕金森病影响的脑区(蓝斑、腹侧延髓、黑质纹状体多巴胺能神经元)的最终细胞损失。在具体目标2中，我们将把这一分析扩展到采用最先进的BAG技术的Mouse Genetics Core中产生的新的小鼠模型。其中包括一个表达Parkin突变的小鼠模型，该突变被证明会导致果蝇中多巴胺能神经元的丧失。这些小鼠的特征将与上述方法相同，然后在项目2和项目3中进行进一步分析。项目1中的研究将与项目5的纵向临床研究平行，这些研究检查帕金森病患者的疾病进展，包括精神和认知共病。他们将提供有关缺陷的时间进程的关键信息和项目2和3的新模型，并在活体哺乳动物系统中测试中心项目4的细胞模型中产生的假说。

DESCRIPTION (provided by applicant): New evidence indicates that distinct mutations cause familial Parkinson's disease (PD) by mechanisms that may also operate in sporadic PD. These new data point to the importance of cell dysfunction preceding cell death and to the involvement of non-dopaminergic neurons in the disease. Accordingly, identifying mechanisms of cellular dysfunction that are common to multiple causes of PD may offer new therapeutic targets to halt or reverse the course of the disease. This renewal application for the UCLA UDALL Parkinson Disease Center of Excellence focuses on studies of progression of dysfunction, in complementary models expressing PD-causing mutations, and in a well characterized patient population. The center consists of 5 projects supported by an administrative core and a mouse genetics core. In the first three projects we propose to continue coordinated multidisciplinary work supported by the current award to characterize the progression of motor and non-motor behavioral anomalies and neuropathology (project 1), anomalies of neurotransmitter release (project 2) and of synaptic function (project 3) in genetic mouse models of PD, including novel models based on BAG technology. These projects will be complemented by the addition of cellular models (project 4) to analyze the mechanisms of cellular dysfunction leading to the phenotypes observed in the mouse. Studying progression of dysfunction will also be the focus of the new patient oriented component of the Center. In this project (project 5), we will conduct clinical longitudinal studies of disease phenotype after diagnosis, including psychiatric and cognitive co-morbidities. This will be coupled to the development and validation of an improved health-related quality of life assessment tool. These patient oriented studies will provide crucial clinical data for future analyses of genetic material from the same patients and for the translation of our basic research efforts into improved patient care. To identify the cellular alterations leading to neurodegeneration in PD, the UCLA UDALL Parkinson Disease Center of Excellence will focus on early manifestations of the disease occurring before the onset of motor symptoms and their progression. Integrating experimental models and clinical studies, the goal of our center is to understand the mechanisms of these cellular dysfunctions in order to spur the development of therapeutic strategies able to stop the disease process. Project 1 Title: Progression of Behavioral and Pathologic Defects Preceding DA Cell Death in Mouse Models of PD PI: Marie-Francoise Chesselet, MD, PhD DESCRIPTION (provided by applicant): New evidence indicates that distinct mutations cause familial Parkinson's disease (PD) by mechanisms that may also operate in sporadic PD. During the current funding period we have identified cellular dysfunction without cell death in mice expressing various mutations known to cause PD in humans. In this renewal application we will test the hypothesis that common mechanisms of dysfunction may be induced by distinct mutations and offer therapeutic targets for treatment at early stages of the disease, before further cell death causes irreversible damage. Project 1. 2. and 3 form the continuation of the current award and will continue to use a multidisciplinary approach to uncover the mechanisms of neuronal dysfunction in existing and novel mouse models. In specific aim 1 of project 1 we will determine the progression of behavioral deficits and neuropathology in already available mice overexpressing alpha-synuclein under different promoters and parkin KO mice. We will use a battery of sensitive motor tests we have developed to assess the motor phenotype of the mice, and will extend this analysis to non-motor behaviors because related symptoms can have a major impact on patient quality of life, as examined in project 5 of the Center. We will use immunohistochemistry to determine the progression of alpha-synuclein pathology and glial activation throughout multiple brain regions known to be affected in PD, and to detect anomalies in the expression of proteins involved in neurotransmitter release and examined in project 4 of the Center. We will use ligand binding and molecular approaches to identify dysregulation of dopaminergic transmission that will be further examined with neurochemical approaches in project 2 and with electrophysiology in project 3. Finally, eventual cell loss in brain regions that are affected in PD (locus coeruleus, ventral medulla, nigrostriatal dopaminergic neurons) will be assessed in older animals with unbiased stereology. In specific aim 2, we will extend this analysis to novel mouse models generated in the Mouse Genetics Core with state of the art BAG technology. These include a mouse model expressing a parkin mutation shown to cause the loss of dopaminergic neurons in Drosophila. These mice will be characterized with the same methods described above, before being further analyzed in projects 2 and 3. Studies in project 1 will parallel longitudinal clinical studies of project 5 that examine disease progression in PD patients, including psychiatric and cognitive comorbidity. They will provide critical information on the time course of the deficits and new models for projects 2 and 3 and test in an in vivo mammalian system the hypotheses generated in cellular models in project 4 of the Center.