Super-Resolution In Vivo Optical Imaging as a Window to Parkinson's Disease Pathogenesis

帕金森病是一种使人衰弱的神经退行性疾病，困扰着全世界500万人，仅在美国每年就花费140亿美元。大多数病例的原因尚不清楚，但遗传和环境因素与风险增加有关。该项目将揭示与帕金森病神经病理学和进展相关的分子事件，从而促进新的治疗干预和更准确的诊断。更一般地说，将开发一种工具，用于鉴定与蛋白质折叠有关的其他中枢神经系统疾病（如阿尔茨海默病和肌萎缩性侧索硬化症）的分子基础，并用于评估这些疾病的新治疗方法。本科生将与研究生一起纳入本项目的各个方面。这项研究将用于增强普渡大学女性工程项目的示范，以及普渡大学春季节期间的脑碗，这两个项目都是针对K-12的。该项目所涉及的实验室将在课堂上向学生开放，研究人员教授这些课程，以说明概念并展示课堂材料与研究的相关性，这是鼓励研究生研究和教育的一个令人兴奋的工具。一种新的适用于活体大脑的基于荧光的光学分子成像方法将被开发和应用，作为一种手段，首次阐明，与帕金森病神经病理学的时间和空间发展有关的致病机制。突触前蛋白α-突触核蛋白的异常蛋白质聚集被认为是关键的早期致病事件，并且聚集被假定首先发生在全身，然后在脑干中，然后在脑的特定区域的后期阶段中，其中神经元损失或功能障碍导致主要运动和非运动症状。聚集体在这些不同大脑区域的传播被认为与疯牛病和克雅氏综合征中朊病毒蛋白的传播相似。了解α-突触核蛋白聚集和帕金森病之间的关系至关重要，以便开发检测和治疗方案。异常的钙信号传导也是一个关键的帕金森病指标，脆弱神经元中的钙离子扰动可以用可用的荧光报告分子监测。拟议的超分辨率光学成像方法将有能力确定α-突触核蛋白聚集和位点进展以及活体动物大脑中的钙信号传导，可能具有10微米的分辨率。研究计划是根据两个目标组织的。在FIRST目标下，将同时进行计算成像和大鼠模型研究，以研究被认为导致帕金森病进展的α-突触核蛋白朊病毒样扩散机制。每周收集的数据将逐步用于帮助确定α-突触核蛋白聚集的精确点位置。这些网站的荧光寿命分布预计也会有所不同，基于聚集水平。根据第二个目标，时间信号信息将从细胞培养物中荧光钙报告分子的显微镜测量中获得，并用于模拟大脑中的神经元激活。这项联合实验和建模工作将建立一个框架，以允许调查钙信号的扰动是否是帕金森病神经元功能障碍或丧失的基础，以及钙和α-该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查进行评估，被认为值得支持的搜索.

Parkinson's disease is a debilitating neurodegenerative disorder afflicting 5 million people worldwide, costing $14 billion a year in the United States alone. The causes of most cases are unknown, but genetic and environmental factors are linked to increased risk. This project will reveal the molecular events related to Parkinson's disease neuropathology and progression, thus facilitating new therapeutic interventions and more accurate prognoses. More generally, a tool will be developed for identifying the molecular basis of other central nervous system disorders related to protein folding, such as Alzheimer's disease and amyotrophic lateral sclerosis, and for evaluating new treatment methods for these diseases. Undergraduate students will be incorporated into aspects of this project with graduate students. The research will be used to enhance demonstrations for the Purdue Women in Engineering program and at the Brain Bowl during Purdue's Spring Fest, both of which target K-12. The laboratories involved on this project will be opened to students in classes that the investigators teach to illustrate concepts and demonstrate the relevance of class material to research, an exciting tool for encouraging graduate research and education.A new fluorescence-based optical molecular imaging method suitable for the living brain will be developed and applied as a means to elucidate, for the first time, pathogenic mechanisms relating to the temporal and spatial development of Parkinson's disease neuropathology. Abnormal protein aggregation of the presynaptic protein alpha-synuclein is thought to be a critical early pathogenic event, and aggregation is postulated to occur first systemically, then in the brain stem, and then in later stages in specific areas of the brain where neuronal loss or dysfunction results in the cardinal motor and non-motor symptoms. The spread of aggregates through these different brain regions is thought to be similar to the spread of the prion protein involved in mad cow disease and Creutzfeldt-Jakob syndrome. Understanding the relationship between alpha-synuclein aggregation and Parkinson's disease is of fundamental importance, in order to develop detection and treatment options. Aberrant calcium signaling is also a key Parkinson's disease indicator, and calcium ion perturbations in vulnerable neurons can be monitored with available fluorescent reporters. The proposed super-resolution optical imaging method will have the capacity to determine alpha-synuclein aggregation and site progression as well as calcium signaling in the brain of a living animal, potentially with a resolution of ten microns. The Research Plan is organized under two aims. Under the FIRST Aim, a simultaneous computational imaging and rat model study will be pursued to investigate the alpha-synuclein prion-like spreading mechanism that is thought to result in Parkinson's disease progression. Data collected weekly will be progressively used to aid the determination of precise point locations for alpha-synuclein aggregation. The fluorescent lifetime distribution of these sites is also expected to differ, based on aggregation level. Under the SECOND Aim, temporal signaling information will be obtained from microscope measurements of fluorescent calcium reporters in cell cultures and used to simulate neuron activation in the brain. This joint experimental and modeling effort will build a framework to allow investigation of whether perturbations of calcium signaling underlie neuron dysfunction or loss in Parkinson's disease, and the hypothesis that interactions between calcium and alpha-synuclein are involved in neurotoxicity.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.