Improved Retinal Organoid Differentiation by Modulating Signaling Pathways Revealed by Comparative Transcriptome Analyses with Development In Vivo
Improved Retinal Organoid Differentiation by Modulating Signaling Pathways Revealed by Comparative Transcriptome Analyses with Development In Vivo
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DOI：
10.1016/j.stemcr.2019.09.009
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发表时间：
2019-11-12
影响因子：
5.9
通讯作者：
Swaroop, Anand
中科院分区：
文献类型：
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作者：
Brooks, Matthew J.;Chen, Holly Y.;Swaroop, Anand
干细胞衍生的视网膜类器官概括了体内分化的许多标志，但缺乏不同细胞类型（尤其是光感受器）的功能成熟。通过全面的时间转录组分析，我们发现，从出生后第 6 天（P6）到 P10 的转录组转变（与小鼠视网膜发育过程中的形态发生和突触形成相关）在类器官中并不明显，并且具有相似模式的共表达簇包含不同的基因组。此外，网络分析发现体内和类器官中视网膜发育之间存在不同的调节动态，其中特定信号通路的时间失调以及涉及光感受器功能和存活的基因表达延迟或减少。因此，在类器官培养物中添加二十二碳六烯酸和成纤维细胞生长因子1可以特异性促进光感受器（包括视锥细胞）的成熟。因此，我们的研究确定了视网膜类器官发育中缺乏的调节信号，并通过在体外产生更成熟的视网膜来提供实验验证，从而促进疾病建模和治疗的研究。
Stem cell-derived retinal organoids recapitulate many landmarks of in vivo differentiation but lack functional maturation of distinct cell types, especially photoreceptors. Using comprehensive temporal transcriptome analyses, we show that transcriptome shift from postnatal day 6 (P6) to P10, associated with morphogenesis and synapse formation during mouse retina development, was not evident in organoids, and co-expression clusters with similar patterns included different sets of genes. Furthermore, network analysis identified divergent regulatory dynamics between developing retina in vivo and in organoids, with temporal dysregulation of specific signaling pathways and delayed or reduced expression of genes involved in photoreceptor function(s) and survival. Accordingly, addition of docosahexaenoic acid and fibroblast growth factor 1 to organoid cultures specifically promoted the maturation of photoreceptors, including cones. Our study thus identifies regulatory signals deficient in developing retinal organoids and provides experimental validation by producing a more mature retina in vitro, thereby facilitating investigations in disease modeling and therapies.