Assembly and characterization of human cortico-striatal neural networks

人类皮质纹状体神经网络的组装和表征

基本信息

批准号：
10629418
负责人：
Oleksandr Shcheglovitov
金额：
$ 53.21万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10629418
关键词：
3-Dimensional Action Potentials Address Affect Anatomy Animal Model Astrocytes Behavior Brain Brain Diseases Brain region Cells Chronic Coculture Techniques Communication Complex Corpus striatum structure Development Diagnostic Disease Disease model Electrodes Electrophysiology (science)Epilepsy Exhibits Genetic Materials Human Huntington Disease Immunohistochemistry Implant Laboratories Major Depressive Disorder Mental disorders Methods Modeling Molecular Molecular Abnormality Mood Disorders Morphology Nervous System Neurons Oligodendroglia Organoids Parkinson Disease Pathologic Pathway interactions Patients Perception Phelan-McDermid syndrome Phenotype Physiological Physiology Population Property Protocols documentation Quantitative Reverse Transcriptase PCR Reproducibility Schizophrenia Science Slice Synapses System Techniques Testing Time Vertebral column With laterality Work autism spectrum disorder brain tissue drug discovery excitatory neuron experimental study functional disability human model human stem cells induced pluripotent stem cell inhibitory neuron innovation insight model development multi-electrode arrays nerve stem cell nervous system disorder neural neural network novel novel strategies novel therapeutics optogenetics patch clamp precision drugs prevent rabies viral tracing self organization single cell mRNA sequencing single-cell RNA sequencing stem cells system architecture

项目摘要

Disruption of the cortico-striatal network has been found in many human brain disorders, including Parkinson’s disease, Huntington’s disease, autism, schizophrenia, and major depression. Unfortunately, the cellular and molecular deficits responsible for the development of disrupted connectivity in these disorders are very difficult to study because of the limited access to primary human brain tissue and inability to recapitulate disease-related network deficits in animal models. This is a major problem that prevents the discovery of novel therapies for patients. Thus, the objectives of this study are to develop the first robust method for generating human cortico- striatal organoids that recapitulate cortico-striatal connectivity and to use this system to investigate the cellular and molecular mechanisms responsible for the establishment and maturation of human cortico-striatal networks. To this end, we will use our new method to generate cortical and, for the first time, striatal organoids from human stem cell-derived single neural rosettes (SNRs). In our preliminary experiments, we found that SNR-derived cortical organoids consist of different subtypes of pallial neural progenitors, deep and superficial layer cortical excitatory neurons, a small fraction of inhibitory neurons with lateral ganglionic eminence (LGE)-like identities, astrocytes, and oligodendrocytes, whereas SNR-derived striatal organoids are composed of different subtypes of subpallial neural progenitors, D1/D2 medium spiny neurons, a large fraction of inhibitory neurons with LGE- like identities, astrocytes, and oligodendrocytes. In addition, we demonstrated that neurons in 5-month-old SNR- derived organoids show functional and morphological evidence of maturity—firing repetitive action potentials, receiving excitatory and inhibitory synaptic inputs, and exhibiting elaborate dendritic branches and spines. Our specific aims in this study are (1) to develop a robust and reproducible protocol for assembling cortico-striatal organoids with well-defined cell composition and organization; (2) to characterize the establishment of anatomical and functional networks in human cortico-striatal organoids; and (3) to determine the molecular and functional properties of the cortical and striatal neurons that make the connections. Importantly, we will use “cutting-edge” techniques such as single-cell mRNA sequencing, chronically implanted multi-electrode probes, rabies virus tracing, and optogenetics to investigate the underlying cellular and molecular mechanisms. In summary, this project will develop the first protocol for assembling functional human cortico-striatal networks and provide novel insights into the cellular and molecular mechanisms of network connectivity. As most brain disorders impact several brain regions and disrupt interregional brain communication, the ability to generate organoids representing multiple brain regions that replicate the complex nervous system architecture and physiology will constitute a major breakthrough in brain disease modeling and drug discovery.

在许多人类大脑疾病中都发现了皮质纹状体网络的破坏，包括帕金森病不幸的是，细胞和疾病。导致这些疾病中连通性破坏的分子缺陷是非常困难的由于原始人脑组织的获取有限且无法重现与疾病相关的情况而进行研究动物模型中的网络缺陷是阻碍新疗法发现的一个主要问题。因此，本研究的目标是开发第一个可靠的方法来生成人类皮质- 纹状体类器官概括了皮质纹状体的连接性并使用该系统来研究细胞以及负责人类皮质纹状体网络建立和成熟的分子机制。为此，我们将使用我们的新方法从人类中生成皮质类器官，并首次生成纹状体类器官在我们的初步实验中，我们发现 SNR 衍生的。皮质类器官由不同亚型的大脑皮层神经祖细胞、深层和浅层皮质组成兴奋性神经元，一小部分具有外侧神经节隆起（LGE）样特征的抑制性神经元，星形胶质细胞和少突胶质细胞，而 SNR 衍生的纹状体类器官由不同的亚型组成大脑皮层下神经祖细胞、D1/D2 中型多棘神经元、大部分具有 LGE- 的抑制性神经元此外，我们还证明了 5 个月大的神经元 SNR-。衍生的类器官显示出成熟的功能和形态证据——激发重复动作电位，接收兴奋性和抑制性突触输入，并表现出复杂的树突分支和脊柱。本研究的具体目标是（1）开发一种稳健且可重复的组装皮质纹状体的协议 (2) 表征具有明确细胞组成和组织的类器官的建立；人类皮质纹状体类器官的解剖和功能网络；以及（3）确定分子和重要的是，我们将使用建立连接的皮质和纹状体神经元的功能特性。 “尖端”技术，如单细胞 mRNA 测序、长期植入的多电极探针、狂犬病病毒追踪和光遗传学研究潜在的细胞和分子机制。总之，该项目将开发第一个用于组装功能性人类皮质纹状体网络的协议，以及与大多数大脑一样，为网络连接的细胞和分子机制提供了新的见解。疾病会影响多个大脑区域并扰乱区域间的大脑交流，即产生代表多个大脑区域的类器官，复制复杂的神经系统架构生理学将构成脑疾病建模和药物发现的重大突破。