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、同步和门控大脑活动中发挥着关键作用 通过抑制其他神经元。它们的功能障碍，尤其是内侧神经节隆起 (MGE) 衍生的功能障碍 cINs 与各种神经发育性脑部疾病有关，例如精神分裂症 (SCZ) 和 自闭症谱系障碍（ASD）。考虑到人类大脑和啮齿动物大脑之间的差异 大脑导致许多在啮齿动物身上验证的中枢神经系统（CNS）疗法失败 模型中，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 脑部疾病。人类胎儿脑组织无法用于机制研究，但我们开发了一种 从多能干细胞中有效产生同质 MGE 型人类 cIN 群体的方法 健康或患病受试者的（PSC）。我们对它们进行了广泛的表征并展示了它们 真实性和功能性，使得研究复杂的聚合功能后果成为可能 真实患者神经元的遗传学，由于缺乏保守性而无法在小鼠神经元中进行研究 非编码区，其中存在大多数风险位点。然而，体外培养的神经元缺乏其他关键的 大脑环境的组成部分，例如星形胶质细胞、少突胶质细胞、小胶质细胞和血管， 这会显着影响它们的功能。人们一直在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重现体内生理环境，但也有 它们对体内情况的模拟程度仍然有限​​。为了解决这个问题，我们之前 研究中，我们开创了人类神经元-小鼠大脑嵌合体来研究人类 SCZ 神经元的功能 生理环境。尽管我们能够成功识别 SCZ cIN 内在连接 由于小鼠大脑存在缺陷，我们无法分析移植神经元对大脑回路的影响 由于移植小鼠中存在健康的小鼠神经元而导致的行为。因此，在这项拟议的研究中，我们 将在 NodScid gamma (NSG) 小鼠中进行大脑区域特异性 cIN 消融，然后替换 用人类 cIN 消除宿主 cIN，生成区域特异性人源化 cIN 嵌合体。基于之前的 包括我们在内的研究表明，移植的人类成功恢复了受损的小鼠抑制作用 cINs，这些小鼠将使我们能够分析移植的人类 cINs 对大脑回路的功能影响 体内生理环境中的行为。这种新颖的生理模型系统将帮助我们区分 复杂脑部疾病的细胞类型和脑区域特异性疾病机制，并有助于发展 新疗法。