Elucidating the molecular mechanisms behind human neurodevelopmental disorders using brain organoids

利用脑类器官阐明人类神经发育障碍背后的分子机制

• 批准号: 10574589
• 负责人: BENNETT G NOVITCH
• 金额: $ 71.92万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-16 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10574589
• 关键词: Address; Anatomy; Anticonvulsants; Appearance; Brain; Brain region; Calcium; Cells; Complex; Coupling; Culture Techniques; DNA Sequence Alteration; Defect; Development; Disease; Disease model; Electrophysiology (science); Embryo; Exhibits; Frequencies; Functional disorder; Gene Mutation; Genes; Goals; Growth; High Frequency Oscillation; Human; Image; Link; Methods; Methyl-CpG-Binding Protein 2; Microcephaly; Modeling; Molecular; Mosaicism; Mutation; Nature; Nervous System Physiology; Neurodevelopmental Disorder; Neurologic; Neuronal Dysfunction; Neurons; Neurophysiology - biologic function; Organoids; Pathologic; Pathology; Pathway interactions; Patients; Phenotype; Population; Reporting; Rett Syndrome; SCN8A gene; Sampling; Seizures; Severities; Slice; Structural defect; Structure; Symptoms; Syndrome; System; Testing; Therapeutic; Variant; Work; X Chromosome; Zika Virus; clinical phenotype; epileptic encephalopathies; epileptiform; excitatory neuron; experimental study; global health; induced pluripotent stem cell; inhibitory neuron; lissencephaly; mosaic; nervous system disorder; network dysfunction; neural; neural circuit; neural network; neuropathology; neuropsychiatric disorder; neuroregulation; novel therapeutics; pathogen; preservation; protein function; sensor; stem; stem cell model; therapeutic development; tool

PROJECT SUMMARY Neurodevelopmental and neuropsychiatric disorders are a global health problem; yet remarkably little is known about their neurological basis in humans. Consequently, treatment options remain limited. The advent of methods to direct the formation of neurons from human embryonic and induced pluripotent stem cells (collectively hPSCs) provides unprecedented opportunities to both investigate how the function of human neural circuits is subverted by neurological disease and screen for new therapies. A major step towards these goals has been realized by the development of organoid culture techniques through which hPSC can be directed to form spatially organized, brain-like structures. Thus far, brain organoids have been successfully employed to model the impact of genetic mutations and environmental pathogens that result in overt defects in brain growth. However, overall brain structure is preserved in most neurological disorders, and defects are primarily defined by alterations in neural activities. Major challenges thus remain in developing means for defining the organization and function of neural networks within organoids and using this approach to explore underlying disease mechanisms and therapeutic opportunities. In our recent work, we discovered that remarkably complex neural network activities can emerge through the creation of cortex-ganglionic eminence fusion organoids, which permits the intermixing and functional coupling of excitatory and inhibitory neurons. Using a combination of calcium sensor imaging and electrophysiological approaches, we identified that fusion organoids exhibit sustained multifrequency neural oscillations reminiscent of higher network functions seen in intact brain samples and slice cultures. We further developed a fusion organoid model for the neurodevelopmental disorder Rett syndrome and found that organoids harboring mutations in the MECP2 gene exhibit markedly abnormal neural network activities including episodes of hypersynchronous bursting, loss of low-to-mid frequency oscillatory rhythms, and abnormal appearance of epileptiform high frequency oscillations. Together, these studies illustrate the extraordinary potential for the fusion organoid platform to report both normal and dysfunctional neural network functions and recapitulate salient pathological features seen in Rett patients such as seizures. Here, we seek to address three central questions for elucidating the mechanisms underlying neural network dysfunction associated with Rett syndrome and other neurodevelopmental disorders. First, does neural network dysfunction seen in Rett syndrome organoids generated from patients harboring different mutations correlate with the nature of the mutation? Second, what is the impact of cellular mosaicism in MECP2 function on neural network activities? Third, do organoid models for different neurological diseases with a seizure component exhibit shared or distinct network dysfunction profiles? Through our studies, we will explore how brain organoids can be best utilized to determine the root causes of a range of human neuropathologies and work towards the goal of discovering new treatments.

项目摘要 神经发育和神经精神疾病是全球健康问题。然而，很少有人知道 关于他们在人类中的神经基础。因此，治疗方案仍然有限。出现 指导人类胚胎和诱导多能干细胞形成神经元的方法 （统称HPSC）提供了前所未有的机会来研究人类的功能 神经回路被神经系统疾病和新疗法的筛查颠覆。迈向这些的主要一步 通过开发了HPSC的器官培养技术，已经实现了目标 指示形成空间组织的脑状结构。到目前为止，脑器官已成功 用于建模基因突变和环境病原体的影响，从而导致明显的缺陷 大脑生长。但是，整体大脑结构在大多数神经系统疾病中都保留了，缺陷是 主要由神经活动的改变定义。因此，主要的挑战仍然存在于发展的手段 定义器官中神经网络的组织和功能，并使用这种方法探索 潜在的疾病机制和治疗机会。在我们最近的工作中，我们发现 通过创建皮质围绕的杰出，可以出现明显复杂的神经网络活动 融合器官，允许兴奋性和抑制性神经元的混合和功能耦合。 使用钙传感器成像和电生理方法的组合，我们确定了融合 器官表现出持续的多频神经振荡，让人联想到较高的网络功能 完整的大脑样品和切片培养物。我们进一步开发了一个融合器官模型 神经发育障碍RETT综合征，发现在MECP2中携带突变的器官 基因表现出明显异常的神经网络活动，包括超同步爆发的发作， 低到中间的频率振荡节奏的丧失和癫痫样高频的异常外观 振荡。这些研究一起说明了融合器官平台的非凡潜力 报告正常和功能障碍的神经网络功能并概括显着的病理特征 在癫痫发作之类的RETT患者中看到。在这里，我们试图解决三个中心问题，以阐明 与RETT综合征和其他相关的神经网络功能障碍的机制 神经发育障碍。首先，在RETT综合征器官中看到的神经网络功能障碍是否存在 由具有不同突变的患者产生的与突变的性质相关？第二，什么 细胞镶嵌物对MECP2功能对神经网络活动的影响？第三，做类型模型 对于具有癫痫发作的不同神经疾病，具有共享或不同的网络功能障碍 个人资料？通过我们的研究，我们将探讨如何最好地利用脑器官来确定根 一系列人类神经病理学的原因，并致力于发现新疗法的目标。