Defining developmental trajectories of prosensory cells in human inner ear organoids at single-cell resolution.
Defining developmental trajectories of prosensory cells in human inner ear organoids at single-cell resolution.
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DOI：
10.1242/dev.201071
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发表时间：
2023-06-15
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内耳感觉上皮细胞包括机械敏感毛细胞和支持细胞。这两种细胞类型都来自表达SOX 2的前感觉细胞，但这些细胞谱系多样化的机制仍不清楚。为了确定前感觉细胞的转录轨迹，我们使用CRISPR/Cas9建立了SOX 2 -2A-ntdTomato人胚胎干细胞系，并在分化第20天和第60天之间的不同时间点用从内耳类器官分离的SOX 2阳性细胞进行单细胞RNA测序分析。我们的pseudotime分析表明，前庭II型毛细胞主要来自支持细胞，而不是bifated prosensory细胞类器官。此外，离子通道和离子转运蛋白相关基因集在支持细胞中富集，而在前感觉细胞中富集，而Wnt信号相关基因集在毛细胞中富集，而在支持细胞中富集。这些发现提供了有价值的见解如何prosensory细胞在人类内耳发育过程中产生毛细胞和支持细胞，并可能提供线索，以促进毛细胞再生从居民支持细胞在听力损失或平衡障碍的个人。 人内耳类器官的纵向scRNA-seq分析鉴定了支持细胞作为毛细胞的可能前体，以及毛细胞分化期间先前未识别的早期基因的上调。
The inner ear sensory epithelia contain mechanosensitive hair cells and supporting cells. Both cell types arise from SOX2-expressing prosensory cells, but the mechanisms underlying the diversification of these cell lineages remain unclear. To determine the transcriptional trajectory of prosensory cells, we established a SOX2-2A-ntdTomato human embryonic stem cell line using CRISPR/Cas9, and performed single-cell RNA-sequencing analyses with SOX2-positive cells isolated from inner ear organoids at various time points between differentiation days 20 and 60. Our pseudotime analysis suggests that vestibular type II hair cells arise primarily from supporting cells, rather than bi-fated prosensory cells in organoids. Moreover, ion channel- and ion-transporter-related gene sets were enriched in supporting cells versus prosensory cells, whereas Wnt signaling-related gene sets were enriched in hair cells versus supporting cells. These findings provide valuable insights into how prosensory cells give rise to hair cells and supporting cells during human inner ear development, and may provide a clue to promote hair cell regeneration from resident supporting cells in individuals with hearing loss or balance disorders. Longitudinal scRNA-seq analysis of human inner ear organoids identifies supporting cells as possible precursors of hair cells and upregulation of previously unrecognized early genes during hair cell differentiation.