Otic Guidance

项目总结/摘要 这项研究计划的第一个目的是使用单细胞转录组学来描述基因的变化， 在多能干细胞的耳引导过程中发生的表达。它建立在已知的原则之上 内耳发育，并利用单细胞RNA-Seq和计算建模来提取 细胞向非神经外胚层、前基板外胚层和最终耳外胚层的分化途径 诱导和感觉毛细胞谱系。在第二个目标中，建议比较研究 当胚胎干细胞的耳引导失败时，这是一个常见的问题。的 从胚胎干细胞中产生内耳细胞是一项不可靠的事业，阻碍了研究 十几年来成功引导的基因表达变化的计算比较 内耳谱系细胞的谱系偏离了耳分化，将导致鉴定特异性 导致细胞分化失败的信号通路。基于计算建模，可测试 将产生假设并进行实验验证。拟议研究的最终目标是 消除来自干细胞的耳引导的实验变化以建立可靠的耳引导 指导协议，可广泛用于许多实验室。

Project Summary / Abstract The first aim of this research proposal is to use single-cell transcriptomics to describe the changes in gene expression that happen during otic guidance from pluripotent stem cells. It builds on known principles of inner ear development and utilizes single-cell RNA-Seq and computational modeling for extracting the differentiation pathway of cells towards non-neural ectoderm, pre-placodal ectoderm, and ultimately otic induction and the sensory hair cell lineage. In a second aim, it is proposed to comparatively investigate the distinct steps when otic guidance from embryonic stem cells fails, which is a common problem. The generation of inner ear cells from embryonic stem cells is an unreliable enterprise and has hampered research for more than a decade. Computational comparisons of gene expression changes in successfully guided inner ear lineage cells with lineages that stray from otic differentiation will result in the identification of specific signaling pathways that contribute to cell differentiation failures. Based on computational modeling, testable hypotheses will be generated and experimentally validated. The ultimate goal of the proposed research is the elimination of experimental variations of otic guidance from stem cells towards establishing a reliable otic guidance protocol that can be widely utilized by many laboratories.