PDF(Pubmed)
Abstract：
UNASSIGNED: Neural induction of human induced pluripotent stem cells represents a critical switch in cell state during which pluripotency is lost and commitment to a neural lineage is initiated. Although many of the key transcription factors involved in neural induction are known, we know little of the temporal and causal relationships that are required for this state transition.
UNASSIGNED: Here, we have carried out a longitudinal analysis of the transcriptome of human iPSCs undergoing neural induction. Using the temporal relationships between the changing profile of key transcription factors and subsequent changes in their target gene expression profiles, we have identified distinct functional modules operative throughout neural induction.
UNASSIGNED: In addition to modules that govern loss of pluripotency and gain of neural ectoderm identity, we discover other modules governing cell cycle and metabolism. Strikingly, some of these functional modules are retained throughout neural induction, even though the gene membership of the module changes. Systems analysis identifies other modules associated with cell fate commitment, genome integrity, stress response and lineage specification. We then focussed on OTX2, one of the most precociously activated transcription factors during neural induction. Our temporal analysis of OTX2 target gene expression identified several OTX2 regulated gene modules representing protein remodelling, RNA splicing and RNA processing. Further CRISPRi inhibition of OTX2 prior to neural induction promotes an accelerated loss of pluripotency and a precocious and aberrant neural induction disrupting some of the previously identified modules.
UNASSIGNED: We infer that OTX2 has a diverse role during neural induction and regulates many of the biological processes that are required for loss of pluripotency and gain of neural identity. This dynamical analysis of transcriptional changes provides a unique perspective of the widespread remodelling of the cell machinery that occurs during neural induction of human iPSCs.
UNASSIGNED: Here, we have carried out a longitudinal analysis of the transcriptome of human iPSCs undergoing neural induction. Using the temporal relationships between the changing profile of key transcription factors and subsequent changes in their target gene expression profiles, we have identified distinct functional modules operative throughout neural induction.
UNASSIGNED: In addition to modules that govern loss of pluripotency and gain of neural ectoderm identity, we discover other modules governing cell cycle and metabolism. Strikingly, some of these functional modules are retained throughout neural induction, even though the gene membership of the module changes. Systems analysis identifies other modules associated with cell fate commitment, genome integrity, stress response and lineage specification. We then focussed on OTX2, one of the most precociously activated transcription factors during neural induction. Our temporal analysis of OTX2 target gene expression identified several OTX2 regulated gene modules representing protein remodelling, RNA splicing and RNA processing. Further CRISPRi inhibition of OTX2 prior to neural induction promotes an accelerated loss of pluripotency and a precocious and aberrant neural induction disrupting some of the previously identified modules.
UNASSIGNED: We infer that OTX2 has a diverse role during neural induction and regulates many of the biological processes that are required for loss of pluripotency and gain of neural identity. This dynamical analysis of transcriptional changes provides a unique perspective of the widespread remodelling of the cell machinery that occurs during neural induction of human iPSCs.
摘要：
■人诱导多能干细胞的神经诱导代表细胞状态的关键转变,在该转变过程中多能性丧失并且开始对神经谱系的承诺。尽管许多参与神经诱导的关键转录因子是已知的，我们对这种状态转换所需的时间和因果关系知之甚少。
■这里，我们对经历神经诱导的人iPSCs的转录组进行了纵向分析。利用关键转录因子的变化谱与其靶基因表达谱的后续变化之间的时间关系，我们已经确定了在整个神经诱导过程中有效的不同功能模块。
■除了控制多能性丧失和神经外胚层身份获得的模块之外，我们发现了其他控制细胞周期和新陈代谢的模块。引人注目的是，其中一些功能模块在整个神经诱导过程中保留下来，即使模块的基因成员发生了变化。系统分析确定了与细胞命运承诺相关的其他模块，基因组完整性,应激反应和谱系规范。然后，我们专注于OTX2，这是神经诱导过程中最早熟激活的转录因子之一。我们对OTX2靶基因表达的时间分析确定了几个OTX2调节的基因模块，代表蛋白质重塑，RNA剪接和RNA加工。在神经诱导之前对0TX2的进一步CRISPRi抑制促进了多能性的加速丧失和破坏一些先前鉴定的模块的早熟和异常神经诱导。
■我们推断OTX2在神经诱导过程中具有不同的作用，并调节丧失多能性和获得神经同一性所需的许多生物过程。转录变化的这种动力学分析提供了在人iPSC的神经诱导过程中发生的细胞机制的广泛重塑的独特视角。
■这里，我们对经历神经诱导的人iPSCs的转录组进行了纵向分析。利用关键转录因子的变化谱与其靶基因表达谱的后续变化之间的时间关系，我们已经确定了在整个神经诱导过程中有效的不同功能模块。
■除了控制多能性丧失和神经外胚层身份获得的模块之外，我们发现了其他控制细胞周期和新陈代谢的模块。引人注目的是，其中一些功能模块在整个神经诱导过程中保留下来，即使模块的基因成员发生了变化。系统分析确定了与细胞命运承诺相关的其他模块，基因组完整性,应激反应和谱系规范。然后，我们专注于OTX2，这是神经诱导过程中最早熟激活的转录因子之一。我们对OTX2靶基因表达的时间分析确定了几个OTX2调节的基因模块，代表蛋白质重塑，RNA剪接和RNA加工。在神经诱导之前对0TX2的进一步CRISPRi抑制促进了多能性的加速丧失和破坏一些先前鉴定的模块的早熟和异常神经诱导。
■我们推断OTX2在神经诱导过程中具有不同的作用，并调节丧失多能性和获得神经同一性所需的许多生物过程。转录变化的这种动力学分析提供了在人iPSC的神经诱导过程中发生的细胞机制的广泛重塑的独特视角。
