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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要神经发育和神经精神疾病是一个全球性的健康问题；然而我们却知之甚少关于它们在人类中的神经学基础。因此，治疗选择仍然有限。的出现指导人类胚胎干细胞和诱导多能干细胞形成神经元的方法（统称 hPSC）为研究人类功能如何发挥作用提供了前所未有的机会神经回路被神经系统疾病破坏并寻找新疗法。朝着这些目标迈出的重要一步类器官培养技术的发展已经实现了目标，通过该技术可以将 hPSC 旨在形成空间组织的、类似大脑的结构。目前，脑类器官已成功用于模拟基因突变和环境病原体的影响，这些影响会导致明显的缺陷大脑生长。然而，在大多数神经系统疾病中，大脑的整体结构得到了保留，并且缺陷主要由神经活动的改变来定义。因此，主要挑战仍然是开发手段定义类器官内神经网络的组织和功能，并使用这种方法来探索潜在的疾病机制和治疗机会。在我们最近的工作中，我们发现通过皮层神经节隆起的创建可以出现非常复杂的神经网络活动融合类器官，允许兴奋性和抑制性神经元的混合和功能耦合。通过结合钙传感器成像和电生理学方法，我们确定了融合类器官表现出持续的多频率神经振荡，让人想起在完整的大脑样本和切片培养物。我们进一步开发了融合类器官模型神经发育障碍 Rett 综合征，并发现含有 MECP2 突变的类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，中低频率振荡节律丧失，以及癫痫样高频异常出现振荡。这些研究共同证明了融合类器官平台的巨大潜力报告正常和功能失调的神经网络功能并概括显着的病理特征见于 Rett 患者，如癫痫发作。在这里，我们试图解决三个核心问题，以阐明与 Rett 综合征和其他疾病相关的神经网络功能障碍的机制神经发育障碍。首先，雷特综合征类器官中是否存在神经网络功能障碍？由携带不同突变的患者产生的结果与突变的性质相关吗？二、什么 MECP2 功能中的细胞嵌合对神经网络活动有影响吗？三、做类器官模型对于具有癫痫发作成分的不同神经系统疾病，表现出共同或不同的网络功能障碍个人资料？通过我们的研究，我们将探索如何最好地利用大脑类器官来确定根源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。