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

超分辨率体内光学成像作为帕金森病发病机制的窗口

• 批准号: 1937986
• 负责人: Kevin Webb
• 金额: $ 40万
• 依托单位: Purdue University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-15 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1937986&HistoricalAwards=false
• 关键词: Super; Resolution; Vivo; Optical; Imaging

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.

帕金森病是一种使人衰弱的神经退行性疾病，全世界有 500 万人受其困扰，仅在美国每年就造成 140 亿美元的损失。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措施和更准确的预后。更一般地说，将开发一种工具来识别与蛋白质折叠相关的其他中枢神经系统疾病（例如阿尔茨海默病和肌萎缩侧索硬化症）的分子基础，并评估这些疾病的新治疗方法。本科生将与研究生一起参与该项目的各个方面。该研究将用于加强普渡大学女性工程项目和普渡大学春季节期间 Brain Bowl 的演示，这两个项目都针对 K-12。参与该项目的实验室将向研究人员教授的课程中的学生开放，以阐明概念并展示课程材料与研究的相关性，这是鼓励研究生研究和教育的令人兴奋的工具。将开发并应用一种适用于活体大脑的新的基于荧光的光学分子成像方法，作为首次阐明与帕金森病神经病理学时空发展相关的致病机制的手段。突触前蛋白α-突触核蛋白的异常蛋白聚集被认为是关键的早期致病事件，并且推测聚集首先发生在全身，然后发生在脑干，然后在后期发生在大脑的特定区域，其中神经元丢失或功能障碍导致主要运动和非运动症状。聚集体通过这些不同大脑区域的传播被认为与疯牛病和克雅氏综合症中涉及的朊病毒蛋白的传播相似。了解 α-突触核蛋白聚集与帕金森病之间的关系对于开发检测和治疗方案至关重要。异常的钙信号传导也是帕金森病的一个关键指标，并且可以使用可用的荧光报告基因来监测脆弱神经元中的钙离子扰动。所提出的超分辨率光学成像方法将能够确定活体动物大脑中的 α-突触核蛋白聚集和位点进展以及钙信号传导，分辨率可能达到 10 微米。该研究计划有两个目标。在第一个目标下，将进行同步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传播机制。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。根据聚集水平，这些位点的荧光寿命分布预计也会有所不同。在第二个目标下，时间信号信息将从细胞培养物中荧光钙报告基因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，以调查钙信号传导的扰动是否是帕金森病神经元功能障碍或损失的基础，以及钙和 α-突触核蛋白之间的相互作用与神经毒性有关的假设。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响进行评估，被认为值得支持 审查标准。

项目成果

Localization of Fluorescent Targets in Deep Tissue With Expanded Beam Illumination for Studies of Cancer and the Brain

• DOI: 10.1109/tmi.2020.2972200
• 发表时间: 2020-07-01
• 期刊: IEEE TRANSACTIONS ON MEDICAL IMAGING
• 影响因子: 10.6
• 作者: Bentz, Brian Z.;Mahalingam, Sakkarapalayam M.;Webb, Kevin J.
• 通讯作者: Webb, Kevin J.

Fluorescence Lifetime Imaging of Protein Aggregation to Understand the Etiology of Neurodegenerative Diseases

蛋白质聚集的荧光寿命成像以了解神经退行性疾病的病因学

• 发表时间: 2022
• 期刊: Conference on Lasers and Electro-Optics,
• 作者: P.-M. E. Ivey, A. Krishnamoorthi

Two C-terminal sequence variations determine differential neurotoxicity between human and mouse α-synuclein

• DOI: 10.1186/s13024-020-00380-w
• 发表时间: 2020-09-08
• 期刊: MOLECULAR NEURODEGENERATION
• 影响因子: 15.1
• 作者: Landeck, Natalie;Strathearn, Katherine E.;Rochet, Jean-Christophe
• 通讯作者: Rochet, Jean-Christophe