Generation and molecular characterization of multiple human neural cell types

摘要-科学核心 人类诱导多能干细胞（iPSC）为研究人类组织和 器官发育和发现人类疾病的分子和细胞基础。能够 iPSC可以分化成人体中几乎任何类型的细胞，已被证明对研究 以前无法进入的细胞群，如发育和成熟大脑的神经细胞。这些 在嗜神经病毒如寨卡病毒的研究中， ZIKV和西尼罗河病毒（WNV）。 ZIKV是一种蚊媒黄病毒，据报道目前在26个国家和地区传播， 拉丁美洲和加勒比。虽然ZIKV感染与新生儿和其他儿童的小头畸形有关， 大脑异常，如格林-巴利综合征，ZIKV如何损害大脑发育和功能， 未知西尼罗河病毒是黄病毒属的另一个成员，也可导致严重的神经系统疾病，如 脑炎或脑膜炎，其潜在的机制仍然是难以捉摸的，没有报道的研究 使用人类细胞模型。由于进入人脑组织的途径非常有限， 使用人类iPSC的模型可以促进生物发现和药物开发。最近开发 从iPSCs生成3D脑类器官的技术有望实现更准确的 这些模型概括了人类大脑发育的关键过程和特征。ZIKV和WNV建模 中枢神经系统感染，科学核心将优化和标准化的协议， 分化疾病相关的特定细胞类型（特定目的1和2），产生基因修饰的 用于功能研究的iPSC生产线（具体目标1），为所有项目提供细胞并提供现场培训 （特定目标2），并进行生物信息学分析，用于转录组学分析（特定目标3）。的 Scientific Core在为细胞培养的可重现条件设定标准方面发挥着至关重要的作用， 对项目2和3中的人员进行交叉培训，这是强大平台开发的关键组成部分。的 科学核心将与项目1在技术开发和协议优化方面密切合作， 用于记录和传播经验证的协议的管理核心。

SUMMARY – Scientific Core Human induced pluripotent stem cells (iPSCs) offer a remarkable opportunity to study human tissue and organ development and to discover the molecular and cellular basis of human diseases. Capable of being differentiated into nearly any cell type in the human body, iPSCs have proven to be invaluable for studying previously inaccessible cell populations such as neural cells of the developing and mature brain. These populations are among the most relevant cell types in the study of neurotropic viruses such as Zika virus (ZIKV) and West Nile virus (WNV). ZIKV, a mosquito-borne flavivirus, is currently reported to be circulating in 26 countries and territories in Latin America and the Caribbean. While ZIKV infection has been linked to microcephaly in newborns and other brain abnormalities such as Guillain-Barré syndrome, how ZIKV impairs brain development and function is unknown. WNV, another member of flavivirus genus, can also lead to serious neurologic illnesses such as encephalitis or meningitis, with the underlying mechanism remains elusive and there is no reported studies using human cellular models. Since access to human brain tissue is very limited, generating new cellular models using human iPSCs can facilitate biological discovery and drug development. Recently developed technology to generate 3D cerebral organoids from iPSCs holds the promise of allowing for more accurate models that recapitulate key processes and features of human brain development. To model ZIKV and WNV infections of the central nervous system, the Scientific Core will optimize and standardize protocols for differentiation of disease-relevant specific cell types (Specific Aims 1 and 2), generate genetically-modified iPSC lines for functional studies (Specific Aim 1), supply cells and provide on-site training for all projects (Specific Aim 2), and perform bioinformatics analyses for transcriptomic profiling (Specific Aim 3). The Scientific Core plays an essential role in setting standards for reproducible conditions for cell culture through cross-training of personnel in Projects 2 & 3, which is a critical component of robust platform development. The Scientific Core will work closely with Project 1 on technology development and protocol optimization and the Administrative Core for documentation and dissemination of validated protocols.