Cellular and molecular mechanisms of brain dysfunction in NF1

尽管众所周知，1型神经纤维瘤病(NF1)存在认知缺陷，但其背后的机制 神经病理生理学仍不清楚。明显缺乏从小鼠模型的发现到临床的转化 表现出啮齿动物和人类在发育、结构和功能上的固有物种差异 大脑，并强调迫切需要开发和使用人体模型系统来研究人类的各个方面 大脑紊乱，并弥合翻译的差距到临床。我们的长期目标是开发和利用人类 用细胞模型阐明相关认知缺陷的分子和细胞机制 与NF1合作。表观遗传重编程、干细胞生物学和基因工程领域的最新进展 为我们提供了一个独特的机会来模拟神经发育障碍，如NF1 与小鼠转基因方法高度互补，因为它保持了对复杂人类细胞的保真度 上下文。目前这项研究的目的是利用这些工具来表征脑血管畸形 NF1相关突变引起的神经发育和神经元网络活动 神经纤维蛋白1(NF1编码蛋白)调控生物学的细胞和分子机制 在不同的蜂窝环境中的过程。根据申请人实验室提供的初步数据，我们 假设NF1基因的疾病相关突变影响人类的增殖和细胞命运 控制神经元输出的神经前体细胞；并通过潜在的 人类特有的机制。这一假设将使用两个互补的细胞模型进行验证：1)人类 脑有机体将用于研究神经纤维蛋白1在人类神经前体细胞类型中的作用， 尤其是在啮齿动物发育的皮质中大量缺失的外径向神经胶质细胞。一个多学科的 方法将被用来表征神经上皮细胞扩增、迁移、 2)诱导神经元(IN)系统，该系统是由 申请者和其他人将被用来剖析不规则神经元网络背后的细胞机制 在由NF1突变神经元组成的培养物中观察到活性。目前的研究将是第一个系统化的研究 研究神经纤维蛋白1利用人体模型系统完成神经系统中所建议的投影功能。成功将为神经纤维蛋白1在不同神经细胞中的作用提供新的知识 与NF1的病理生理相关的细胞类型。鉴于NF1自闭症的高患病率， 拟议中的研究也有可能阐明关键的分子和细胞机制 特发性自闭症。

Despite the well-recognized cognitive deficits in Neurofibromatosis type 1 (NF1), the mechanisms underlying the neuropathophysiology remain unclear. The clear lack of translation from findings in mouse models to the clinic manifests the inherent species differences in the development, architecture and function of rodent and human brains, and underscores the urgent need to develop and use human model systems to study aspects of human brain disorders and to bridge the translational gap to the clinic. Our long-term goal is to develop and use human cellular models to elucidate the molecular and cellular mechanisms underlying the cognitive deficits associated with NF1. Recent advances in the field of epigenetic reprogramming, stem cell biology and genetic engineering has rendered us a unique opportunity to model neurodevelopmental disorders such as NF1 in a manner that is highly complementary to murine transgenic approaches by that maintains fidelity with complex human cellular contexts. The objective of the current study is to harness such tools to characterize the abnormalities in neurodevelopment and neuronal network activity caused by NF1-associated mutations and determine the cellular and molecular mechanisms underlying Neurofibromin 1 (NF1 encoding protein) - regulated biological processes in different cellular contexts. Based on the preliminary data produced in the applicant’s laboratory, we hypothesize that disease associated mutations in NF1 affect proliferation and cell fate commitment of human neural progenitor cells that control neuronal output; and impair neuronal network activity through a potentially human-specific mechanism. The hypothesis will be tested using two complementary cellular models: 1) human brain organoids will be used to investigate the role of Neurofibromin 1 in human neural progenitor cell types, especially the outer radial glial cells that are largely absent in the rodent developing cortices. A multidisciplinary approach will be used to characterize cell type-specific defects in neuroepithelial cell expansion, migration, differentiation, and the mitotic properties of cells; 2) induced neuronal (iN) system, which was developed by the applicant and others, will be used to dissect the cellular mechanisms underlying the irregular neuronal network activity observed in cultures consisting of NF1 mutant neurons. The current study would be the first systematic investigation of Neurofibromin 1 completion of the proposed project function in the neural system using a human model system. Successful will provide novel knowledge on the role of Neurofibromin 1 in different neural cell types that are relevant to the pathophysiology of NF1. Given the high prevalence of Autism in NF1, the proposed research also has the potential to shed light on the key molecular and cellular mechanisms underlying idiopathic Autism.