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: 10538043
• 负责人: Nikki Tjahjono
• 金额: $ 3.92万
• 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/10538043
• 关键词: Alzheimer' s Disease; Basal Ganglia; Biosensor; Brain Diseases; CRISPR-mediated transcriptional activation; Calcium; Cause of Death; Cell Compartmentation; Cell Death; Chemicals; Defect; Development; Devices; Disease; Disease Progression; Disease model; Dyes; Engineering; Failure; Functional Imaging; Functional disorder; Glutamates; Human; Huntington Disease; Hybrids; In Vitro; Individual; Interneurons; Label; Measurement; Measures; Methods; Microfluidic Microchips; Microfluidics; Midbrain structure; Modeling; Molecular; Molecular Disease; Monitor; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Onset of illness; Optics; Outcome; Output; Parkinson Disease; Pathologic; Pathology; Patients; Pharmaceutical Preparations; Pharmacology; 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; base; 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 circuit; neurochemistry; neuropsychiatry; neurotransmitter release; next generation; novel; optical sensor; optogenetics; overexpression; prevent; relating to nervous system; response; sensor; success; synaptic function; therapy outcome; tool; transmission process; Alzheimer' s Disease; Basal Ganglia; Biosensor; Brain Diseases; CRISPR-mediated transcriptional activation; Calcium; Cause of Death; Cell Compartmentation; Cell Death; Chemicals; Defect; Development; Devices; Disease; Disease Progression; Disease model; Dyes; Engineering; Failure; Functional Imaging; Functional disorder; Glutamates; Human; Huntington Disease; Hybrids; In Vitro; Individual; Interneurons; Label; Measurement; Measures; Methods; Microfluidic Microchips; Microfluidics; Midbrain structure; Modeling; Molecular; Molecular Disease; Monitor; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Onset of illness; Optics; Outcome; Output; Parkinson Disease; Pathologic; Pathology; Patients; Pharmaceutical Preparations; Pharmacology; 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; base; 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 circuit; neurochemistry; neuropsychiatry; neurotransmitter release; next generation; novel; optical sensor; optogenetics; overexpression; prevent; relating to nervous system; response; sensor; 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.

项目摘要 缓慢或延迟疾病发作，减轻症状并改善疾病的神经退行性疾病的治疗 迫切需要患者的生活质量。神经退行性疾病的共同特征，例如 帕金森氏病（PD）是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，新兴研究表明，这些过程可能发生在 宽度变性，使其成为有望开发神经保护疗法的目标。 因此，该项目旨在开发新工具，以更好地了解全球神经化学中的早期缺陷 神经变性的动力学，突触生理和电路结构。剥夺前模型 通过遗传操纵内源性α-核蛋白的遗传操纵，零星PD将建立 该疾病的中心标志（AIM 1）。鉴于PD啮齿动物模型的可翻译性局限 项目利用使用微流体在功能电路中组织的人类多能干细胞衍生的神经元 培养装置。该设备不仅允许对α-核蛋白的电路特定操纵，还允许 特定的成熟和组织不同的细胞类型以概括基本的神经节输入。高保真 以及该模型中电路结构和功能变化的特定电路的测量，创新 将追求新颖的光学工具。该提案中开发的光学传感器利用了混合动力 涉及基于遗传释放释放hotag的蛋白质支架和明亮的红色的化学属策略 转移的染料。远红色杂种基于卤素的化学夹谷氨酸传感器将与A一起使用 钙指示器和光遗传执行器，以允许全光操作和读出电路功能 早期PD（AIM 2）。将开发基于拆分释放屏的突触传感器以启用简单 用细胞类型和输入的突触密度和突触谷氨酸传播的变化测量 特异性（目标3）。这些工具将使我们能够测试中心假设，即电路特定的操纵 人皮质 - 基质神经节回路中的α-突触核蛋白将导致谷氨酸活性的不同调节， 突触功能和电路结构。成功完成该项目也将导致 建立其他神经退行性疾病和疗法的通用平台 可以理解干预措施。