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.

摘要 GABA能皮层中间神经元（cINs）在平衡、同步和门控大脑活动中起关键作用 通过抑制其他神经元。他们的功能障碍，特别是那些内侧神经节隆起（MGE）衍生 cINs与各种神经发育性脑障碍有关，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的差异 脑的损伤导致许多在啮齿类动物中验证的中枢神经系统（CNS）疗法失败 因此，研究人类神经元以更好地理解这些cIN相关的神经元的机制是至关重要的。 脑部疾病。人类胎儿脑组织无法进行机制研究，但我们已经开发了一种 从多能干细胞有效产生MGE型人cIN的同质群体的方法 在健康或患病受试者的PSC中。我们已经广泛地描述了它们的特征，并证明了它们的 真实性和功能性，使研究复杂的聚合功能后果成为可能。 在真实的患者神经元中的遗传学，由于缺乏保守性， 非编码区，大多数风险基因座都存在。然而，体外培养的神经元缺乏其他关键的 脑环境的组分，如星形胶质细胞、少突胶质细胞、小胶质细胞和血管， 这会严重影响它们的功能。已经努力优化体外培养系统， 通过添加其他脑细胞成分，可以更好地重现体内生理环境，但 但是它们在模拟体内情况的紧密程度方面仍然存在局限性。为了解决这个问题，在我们以前的 在这项研究中，我们开创了人类神经元-小鼠脑嵌合体的先河，以研究人类SCZ神经元的功能， 生理环境。虽然我们能够成功地识别SCZ cIN-内在连接， 由于小鼠大脑中存在缺陷，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，在这项研究中，我们 将在NodScid gamma（NSG）小鼠中进行脑区域特异性cIN消融，然后替换 用人cIN消融宿主cIN以产生区域特异性人源化cIN嵌合体。基于先前 包括我们在内的研究表明，移植的人 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， 在体内生理环境中的行为。这个新颖的生理模型系统将帮助我们 细胞类型和脑区域特异性疾病机制的复杂的大脑疾病，并帮助发展 新疗法。