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

• 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
• 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

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.

在包括帕金森氏症在内 疾病，亨廷顿病，自闭症，精神分裂症和严重抑郁症。不幸的是，细胞和 分子定义了负责这些疾病中连通性破坏的发展非常困难的 由于获得原发性人脑组织的机会有限而无法概括与疾病有关的研究 网络在动物模型中定义。这是一个主要的问题，可以防止发现新颖的疗法 患者。这是这项研究的目的是开发第一种强大的方法来产生人类皮质 概括皮质 - 纹状体连通性并使用该系统研究细胞的纹状体器官 以及负责人皮质 - 纹状体网络建立和成熟的分子机制。 为此，我们将使用新方法来产生皮质，并首次从人类 干细胞衍生的单神经玫瑰花结（SNR）。在我们的初步实验中，我们发现SNR衍生 皮质器官由不同亚型组成的乳腺神经源性器，深层和浅表皮质组成 兴奋性神经元，一小部分具有侧向神经节显着性（LGE）样身份的抑制性神经元， 星形胶质细胞和少突胶质细胞，而SNR衍生的纹状体是由不同的亚型组成的 d1/d2培养基神经元的少数抑制性神经元的D1/D2培养基神经元的抑制性神经元 像身份，星形胶质细胞和少突胶质细胞一样。此外，我们证明了5个月大的SNR-中的神经元 衍生的类器官显示成熟的功能和形态学证据 接收兴奋性和抑制性突触输入，并表现出精心的树突分支和刺。我们的 这项研究的具体目的是（1）开发一种可重复的协议来组装皮质纹状体 具有明确定义的细胞组成和组织的器官； （2）表征建立 人皮质纹状体器官中的解剖和功能网络； （3）确定分子和 具有连接的皮质和纹状体神经元的功能特性。重要的是，我们将使用 “尖端”技术，例如单细胞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，以研究潜在的细胞和分子机制。在 总而言之，该项目将开发第一个用于组装功能性人体皮层 - 纹状体网络的协议和 提供有关网络连接性的细胞和分子机制的新见解。作为大脑 疾病影响几个大脑区域并破坏区域间交流，产生的能力 代表多个大脑区域的器官复制复杂的神经系统结构和 生理学将构成脑疾病建模和药物发现的重大突破。