Alpha-synuclein biomarkers in Parkinson disease

 描述(由申请人提供)：本申请是为宾夕法尼亚大学(U Penn)神经学讲师Rizwan Akhtar博士颁发的K08职业发展奖。这位候选人正在确立他的职业生涯，成为一名运动障碍和神经退行性疾病领域的神经学家-神经学家。这个为期五年的项目的目标是开发两个基于α-突触核蛋白的新生物标记物，并将这些生物标记物应用于帕金森病(PD)的研究。这位候选人的导师是弗吉尼亚·李博士和约翰·特罗亚诺夫斯基博士，他们是宾夕法尼亚大学神经退行性疾病研究中心(CNDR)的联合主任，以及宾夕法尼亚大学帕金森疾病和运动障碍中心(PD&MDC)主任马修·斯特恩博士。这些人在培养成功的内科科学家方面有着悠久的历史。CNDR是一个多学科的研究实验室，拥有蛋白质生物化学和分子生物学、细胞培养、显微镜、组织病理学、小动物外科手术、药物开发和高通量分析方面的内部专业知识。CNDR和PD&MDC还拥有一个多模式数据库，其中包括临床数据、生物液样本、影像研究和尸检样本，这些数据是通过美国国立卫生研究院资助的几个宾夕法尼亚大学项目拨款培养出来的，候选人将利用这些数据库来实现该项目的目标。这位候选人已经组建了一支由临床和研究导师组成的团队，并制定了一项结构化的培训计划，以发展必要的技能集，以过渡到神经退行性疾病研究的独立性。α-突触核蛋白是一种淀粉样蛋白，被认为是帕金森病发病的核心。当错误折叠、磷酸化或后跨国修饰时，α-突触核蛋白似乎通过采用病理构象而对细胞产生毒性。虽然所有帕金森病患者都有神经元丢失和大脑中含有α-突触核蛋白的蛋白质聚集体，但并不是所有的患者都遵循相同的病程或出现相同的并发症。帕金森病的临床异质性仍然不能用已知的关于潜在的神经病理来解释。目前还没有生物标记物来衡量α-突触核蛋白病理在大脑中的传播，也没有针对帕金森病的治疗方法来改变病程。该项目将检验α-突触核蛋白自身抗体(AIM#1)和病理形式的α-突触核蛋白(AIM#2)是帕金森病的生物标志物的假设。所采取的方法将是为这些生物标记物开发两种基于酶联免疫吸附试验的分析方法，然后在传播性联核病小鼠模型中验证每一种分析方法(目标3)。每个化验都将使用新产生的抗体和截短的、磷酸化的和纤维状的重组α-向斜进行严格的测试。一旦开发出来，这些分析将被用于测试CNDR的血清和脑脊液，并使用多模式方法将每个生物标记物与临床疾病状态相关联。该项目将提供必要的研究技能和培训，使候选人能够将这些生物标记物转化为临床护理，并发展一个专注于帕金森病发病机制的独立研究实验室。

 DESCRIPTION (provided by applicant): This application is for a K08 Career Development Award for Dr. Rizwan Akhtar, who is an Instructor in Neurology at the University of Pennsylvania (U Penn). The candidate is establishing his career as a neurologist - neuroscientist in the fields of movement disorders and neurodegenerative disease. The objective of this five-year project is to develop two new biomarkers based on alpha-synuclein and apply these biomarkers to the study of Parkinson disease (PD). The candidate is mentored by Drs. Virginia Lee and John Trojanowski, who co-direct the Center for Neurodegenerative Disease Research (CNDR) at U Penn, and by Dr. Matthew Stern, Director of the Parkinson Disease and Movement Disorders Center (PD&MDC) at U Penn. These individuals have a long history of training successful physician - scientists. The CNDR is a multi- disciplinary research laboratory with in-house expertise in protein biochemistry and molecular biology, cell culture, microscopy, histopathology, small animal surgery, drug development, and high throughput assays. The CNDR and PD&MDC also have a multimodal database of clinical data, biological fluid samples, imaging studies, and autopsy specimens cultivated via several NIH funded Program Project Grants at U Penn, which the candidate will utilize to achieve the Aims of this project. The candidate has assembled a team of clinical and research mentors and a structured training plan to develop the necessary skill-set to transition to independence in neurodegenerative disease research. Alpha-synuclein is an amyloid genic protein and thought to be central to the pathogenesis of PD. When misfolded, phosphorylated, or post-transnationally modified, alpha-synuclein appears to confer toxicity to cells by adopting pathological conformations. While all patients with PD have neuron loss and protein aggregates that contain alpha-synuclein in the brain, not all patients follow the same disease course or develop the same complications. The clinical heterogeneity of PD remains poorly explained by what is known regarding the underlying neuropathology. There are currently no biomarkers to measure the spread of alpha-synuclein pathology through the brain, and no treatments for PD appear to modify the disease course. This project will test the hypothesis that alpha-synuclein auto-antibodies (Aim #1) and pathologic forms of alpha-synuclein (Aim #2) are biomarkers for PD. The approach taken will be to develop two ELISA based assays for these biomarkers, and then validate each assay in a mouse model of transmissible synucleinopathy (Aim #3). Each assay will be rigorously tested using newly generated antibodies and truncated, phosphorylated, and fibrillary forms of recombinant alpha-syncline. Once developed, these assays will be used to test serum and CSF from CNDR and correlate each biomarker to clinical disease state using a multi-modal approach. This project will provide the necessary research skills and training to allow the candidate to translate these biomarkers to clinical care and to develop an independent research laboratory focused on mechanisms of PD pathogenesis.