Otic Guidance

耳部指导

• 批准号: 10519111
• 负责人: Stefan Heller
• 金额: $ 15.74万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10519111
• 关键词: Affect; Appearance; Applications Grants; Awareness; Biopsy; Cell Differentiation process; Cell Lineage; Cell Separation; Cells; Computer Analysis; Computer Models; Data; Data Analyses; Dissociation; Drug Evaluation; Drug Screening; Ectoderm; Epithelial Cells; Epithelium; Failure; Gene Expression; Generations; Goals; Hair Cells; Individual; Laboratories; Labyrinth; Location; Methanol; Methods; Modification; Mus; Pathway interactions; Patients; Phenotype; Pluripotent Stem Cells; Population; Protocols documentation; Research; Research Proposals; Sensory; Sensory Hair; Signal Pathway; Signal Transduction; Supporting Cell; Technology; Validation; Variant; Vesicle; cell type; comparative; embryonic stem cell; experimental study; improved; inhibitor; inner ear development; inner ear diseases; placodal ectoderm; preservation; sample fixation; single-cell RNA sequencing; stem cells; three dimensional cell culture; transcriptomics

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.

项目摘要 /摘要 该研究建议的第一个目的是使用单细胞转录组学来描述基因的变化 在多能干细胞的氧气中发生的表达。它建立在已知原理的基础上 内耳发育并利用单细胞RNA-seq和计算建模来提取 细胞朝非神经外胚层，抛牙前外胚层和最终眼膜的差异途径 诱导和感觉毛细胞谱系。在第二个目标中，提议相对研究 当来自胚胎干细胞失败的氧气指控失败时，这是一个常见的问题。这 胚胎干细胞的内耳细胞生成是一种不可靠的企业，并且阻碍了研究 十多年来。基因表达的计算比较成功指导 内耳谱系细胞具有谱系，这些谱系会导致眼部差异，将导致特定的鉴定 有助于细胞差异故障的信号通路。基于计算建模，可测试 假设将产生并通过实验验证。拟议研究的最终目标是 消除从干细胞到建立可靠的氧气的氧气变化 许多实验室可以广泛使用的指导协议。