Brain-region-specific humanized cortical interneuron mice

脑区域特异性人源化皮质中间神经元小鼠

• 批准号: 10735991
• 负责人: SANGMI CHUNG
• 金额: $ 66.05万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10735991
• 关键词: Ablation; Address; Area; Astrocytes; Behavior; Behavioral; Behavioral Assay; Biological Models; Blood Vessels; Brain; Brain Diseases; Brain region; Cell Culture Techniques; Central Nervous System; Chimera organism; Complex; Data; Development; Disease; Dorsal; Dose; Electrodes; Electrophysiology (science); Environment; Failure; Future; Genetic; Graft Survival; Hippocampus; Histologic; Human; Immunohistochemistry; In Vitro; Injections; Interneuron function; Interneurons; Medial; Methods; Microglia; Modeling; Mus; Neurons; Oligodendroglia; Pathogenesis; Patients; Phenotype; Physiological; Play; Pluripotent Stem Cells; Population; Prefrontal Cortex; Qualifying; Regulation; Rodent; Rodent Model; Role; Schizophrenia; Slice; Source; Synapses; System; Testing; Therapeutic; Titrations; Transplantation; Untranslated RNA; Viral Vector; autism spectrum disorder; cell type; cognitive function; cognitive task; cognitive testing; disease mechanisms study; experimental study; human disease; human fetal brain tissue; in vivo; in vivo Model; induced pluripotent stem cell; neuropsychiatric disorder; novel; novel therapeutics; optogenetics; physiologic model; restoration; risk variant; stem cells

Abstract GABAergic cortical interneurons (cINs) play critical roles in balancing, synchronizing, and gating brain activity by inhibiting other neurons. Their malfunction, especially those of medial ganglionic eminence (MGE)-derived cINs, has been associated with various neurodevelopmental brain disorders, such as schizophrenia (SCZ) and autism spectrum disorders (ASD). Considering the fact that the divergence between human brains and rodent brains has resulted in the failure of many central nervous system (CNS) therapeutics validated in rodent models, it is critical to study human neurons to better understand the mechanisms of these cIN-associated brain disorders. Human fetal brain tissues are not accessible for mechanistic studies, but we have developed a method to efficiently generate homogeneous populations of MGE-type human cINs from pluripotent stem cells (PSCs) of healthy or diseased subjects. We have extensively characterized them and demonstrated their authenticity and functionality, making it possible to study the converging functional consequences of complex genetics in real patient neurons, which cannot be studied in mouse neurons due to a lack of conservation of non-coding regions, where most of risk loci are present. However, in vitro cultured neurons lack other critical components of the brain environment, such as astrocytes, oligodendrocytes, microglia and blood vessels, which can significantly impact their function. There have been efforts to optimize in vitro culture systems to better recapitulate in vivo physiological environments by adding other brain cellular components, but there are still limitations as to how closely they can simulate in vivo situations. To resolve this issue, in our previous study, we pioneered human neuron-mouse brain chimeras to study the function of human SCZ neurons in physiological environments. Although we were able to successfully identify SCZ cIN-intrinsic connectivity deficits in mouse brains, we were not able to analyze the impacts of grafted neurons on brain circuits and behaviors due to the presence of healthy mouse neurons in the grafted mice. Thus, in this proposed study, we will perform brain-region-specific cIN-ablation in NodScid gamma (NSG) mice, followed by the replacement of ablated host cINs with human cINs to generate region-specific humanized cIN chimeras. Based on previous studies, including ours, that show successful restoration of compromised mouse inhibition by grafted human cINs, these mice will allow us to analyze the functional impacts of grafted human cINs on the brain circuits and behaviors in physiological in vivo environments. This novel physiological model system will help us tease apart cell-type- and brain-region-specific disease mechanisms for complex brain disorders, and aid in developing novel therapeutics.

抽象的 GABA能皮质中间神经元（CIN）在平衡，同步和门控脑活动中起关键作用 通过抑制其他神经元。它们的故障，尤其是内侧神经节显着性（MGE）的故障 CIN与各种神经发育脑部疾病有关，例如精神分裂症（SCZ）和 自闭症谱系障碍（ASD）。考虑到人大脑和啮齿动物之间的差异 大脑导致许多中枢神经系统（CNS）在啮齿动物中验证 模型，研究人类神经元以更好地了解这些CIN相关的机制至关重要 脑疾病。人胎脑组织无法用于机械研究，但我们已经开发了 有效产生来自多能干细胞的MGE型人CIN的均质种群的方法 健康或患病受试者的（PSC）。我们对它们进行了广泛的特征，并证明了他们的 真实性和功能，使得研究复杂的功能后果成为可能 实际患者神经元中的遗传学，由于缺乏保护，无法在小鼠神经元中研究 非编码区域，大多数风险基因座存在。但是，体外培养的神经元缺乏其他关键 大脑环境的成分，例如星形胶质细胞，少突胶质细胞，小胶质细胞和血管， 这可以显着影响其功能。已经努力优化体外培养系统 通过添加其他脑细胞成分，可以更好地概括体内生理环境，但是有 关于它们能够模拟体内情况的近距离，仍然存在局限性。为了解决这个问题，在我们以前的 研究，我们开创了人类神经鼠大脑嵌合体，以研究人类SCZ神经元在 生理环境。尽管我们能够成功识别SCZ CIN-Intrinsic连接性 小鼠大脑的缺陷，我们无法分析移植神经元对脑电路和 由于移植小鼠中健康小鼠神经元的存在引起的行为。因此，在这项拟议的研究中，我们 将在Nodscid伽马（NSG）小鼠中进行大脑区域特异性粉膜，然后更换 带有人类CIN的消融宿主CIN产生特定区域的人性化CIN嵌合体。基于以前的 包括我们的研究表明，嫁接人类的小鼠抑制作用成功地恢复了 CINS，这些小鼠将使我们能够分析移植的人CIN对脑电路的功能影响，并且 生理体内环境中的行为。这个新颖的生理模型系统将有助于我们分开 复杂脑疾病的细胞类型和大脑特异性疾病机制，并有助于发展 新颖的治疗学。