An All Optical Platform for Modeling and Monitoring Early Neurodegeneration in Human Neural Circuits

用于建模和监测人类神经回路早期神经退行性变的全光学平台

• 批准号: 10684701
• 负责人: Nikki Tjahjono
• 金额: $ 4.01万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10684701
• 关键词: Alzheimer' s Disease; Basal Ganglia; Biosensor; Brain Diseases; CRISPR-mediated transcriptional activation; Calcium; Cause of Death; Cell Death; Chemicals; Defect; Development; Devices; Disease; Disease Progression; Disease model; Disparate; Dyes; Engineering; Failure; Functional Imaging; Functional disorder; Ganglia; Glutamates; Human; Huntington Disease; Hybrids; In Vitro; Individual; Label; Measurement; Measures; Methods; Microfluidic Microchips; Microfluidics; Midbrain structure; Modeling; Molecular; Molecular Disease; Monitor; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Onset of illness; Optics; Outcome; Output; Parkinson Disease; Pathologic; Pathology; Patients; Pharmaceutical Preparations; Physiological; Physiology; Process; Proteins; Quality of life; Research; Resolution; Rodent Model; Scaffolding Protein; Solid; Specificity; Structure; Symptoms; Synapses; Synaptic Transmission; System; Testing; Therapeutic; Therapeutic Intervention; World Health Organization; alpha synuclein; burden of illness; calcium indicator; cell type; density; drug development; functional plasticity; genetic manipulation; genome editing; human pluripotent stem cell; immunocytochemistry; improved; innovation; multiplexed imaging; neural; neural circuit; neurochemistry; neuroprotection; neuropsychiatry; neurotransmitter release; next generation; novel; optical sensor; optogenetics; outcome disparities; overexpression; pharmacologic; postsynaptic; presynaptic; prevent; redshift; response; sensor; sporadic Parkinson' s Disease; success; synaptic function; therapy outcome; tool; transmission process

PROJECT SUMMARY Treatments for neurodegenerative disorders that slow or delay disease onset, mitigate symptoms, and improve patient quality of life are desperately needed. A common feature of neurodegenerative diseases such as Parkinson’s Disease (PD) is synaptic dysfunction and excitatory-inhibitory imbalance. Although traditionally seen as consequences of cell death, emerging research has shown that these processes may occur before widespread degeneration, making them promising targets for the development of neuroprotective therapeutics. Therefore, this project aims to develop new tools to better understand early defects in global neurochemical dynamics, synaptic physiology, and circuit structure in neurodegeneration. A pre-degeneration model of sporadic PD will be established in this project by genetic manipulation of endogenous alpha-synuclein, a central hallmark of the disease (Aim 1). Given the limitations in the translatability of rodent models of PD, this project utilizes human pluripotent stem cell derived neurons organized in functional circuits using a microfluidic culture device. This device allows for not only circuit-specific manipulation of alpha-synuclein, but also the specific maturation and organization of distinct cell types to recapitulate basal ganglia inputs. For high-fidelity and circuit-specific measurements of changes in circuit structure and function in this model, innovations in novel optical tools will be pursued. The optical sensors developed in this proposal utilizes a hybrid chemigenetic strategy that involves a genetically encoded HaloTag based protein scaffold and bright, red- shifted dyes. A far-red hybrid HaloTag-based chemigenetic glutamate sensor will be utilized alongside a calcium indicator and optogenetic actuator to allow for all-optical manipulation and read-out of circuit function in early PD (Aim 2). A split HaloTag-based synaptic sensor will be developed to enable simultaneous measurement of changes in synaptic density and synaptic glutamate transmission with cell type and input specificity (Aim 3). These tools will allow us to test the central hypothesis that circuit-specific manipulation of alpha-synuclein in human cortico-basal ganglia circuitry will result in disparate modulation of glutamate activity, synapse function, and circuit structure. Successful completion of this project will also result in the establishment of a general platform from which other neurodegenerative disorders and therapeutic interventions can be understood.

项目总结 治疗神经退行性疾病，延缓或延缓疾病的发生，缓解症状并改善 患者的生活质量是迫切需要的。神经退行性疾病的一个共同特征是 帕金森病(PD)是指突触功能障碍和兴奋性-抑制性失衡。虽然传统上 被视为细胞死亡的后果，新兴的研究表明，这些过程可能发生在 广泛的变性，使它们成为神经保护疗法发展的有希望的靶点。 因此，该项目旨在开发新的工具，以更好地了解全球神经化学的早期缺陷 神经退行性变的动力学、突触生理学和电路结构。一种新的预退化模型 在这个项目中，将通过对内源性α-突触核蛋白的遗传操作来建立散发性帕金森病。 疾病的中心特征(目标1)。考虑到帕金森病啮齿动物模型的可译性限制，这 该项目利用人类多能干细胞来源的神经元，利用微流控技术在功能电路中进行组织 培养装置。这种装置不仅允许对α-突触核蛋白进行电路特异性操作，而且还允许 不同细胞类型的特定成熟和组织，以概括基底神经节的输入。高保真 以及对此模型中电路结构和功能变化的特定电路测量， 新的光学工具将被追求。在这项提案中开发的光学传感器利用了一种混合 一种化学遗传策略，涉及一种基于HaloTag的基因编码蛋白质支架和明亮的红色- 转移的染料。一种基于HaloTag的远红混合化学谷氨酸传感器将与一种 钙指示器和光生致动器，允许全光学操作和读出电路功能 早期帕金森病(AIM 2)。将开发一种基于HaloTag的分离式突触传感器，使同步 突触密度和突触谷氨酸传递随细胞类型和输入的变化 特异性(目标3)。这些工具将允许我们测试特定于电路的操作的中心假设 人类皮质-基底节回路中的α-突触核蛋白将导致谷氨酸活性的不同调节， 突触功能，以及电路结构。这一项目的成功完成还将导致 建立一个通用平台，其他神经退行性疾病和治疗 干预是可以理解的。