Single-cell reconstruction of differentiation trajectory reveals a critical role of ETS1 in human cardiac lineage commitment
Single-cell reconstruction of differentiation trajectory reveals a critical role of ETS1 in human cardiac lineage commitment
复制标题
分化轨迹的单细胞重建揭示了 ETS1 在人类心脏谱系承诺中的关键作用
DOI：
10.1186/s12915-019-0709-6
复制
发表时间：
2019-11-13
期刊：
影响因子：
5.4
通讯作者：
Wang, Li
中科院分区：
文献类型：
--
作者：
Ruan, Hang;Liao, Yingnan;Wang, Li
人类多能干细胞的心脏分化为体外研究人类心脏发育提供了独特的机会，并为心脏再生提供了潜在的细胞来源。与大量研究心脏成熟和心肌细胞亚型特异性诱导的研究相比，多能干细胞早期心脏谱系承诺的分子事件仍然缺乏特征。结果为了揭示多能分化过程中控制心脏谱系承诺的关键分子事件和调节因子，我们对6879个人胚胎干细胞心脏分化过程中6个阶段的细胞进行了单细胞RNA-Seq测序，获得了高质量的数据，并鉴定了具有不同分子特征的多个细胞亚群。通过构建心脏分化的发育轨迹和假定的配体-受体相互作用，我们揭示了心脏祖细胞和内胚层细胞之间的串扰，这可能为第5天心脏谱系承诺提供潜在的细胞微环境。此外，单细胞RNA-Seq数据的计算分析揭示了ETS1 （ETS原癌基因1）激活是由心脏祖细胞和内胚层细胞之间的串扰诱导的重要下游事件。与单细胞分析的结果一致，针对ETS1的染色质免疫沉淀和高通量测序（ChIP-Seq）显示，在第9天和第14天，ETS1在心脏结构基因上的基因组占用，而ETS1的缺失显著损害了心脏分化。总之，我们的研究不仅表征了不同细胞类型的分子特征，并确定了ETS1是细胞间串扰诱导的关键因子，有助于多能状态下的心脏谱系承诺，而且可能对理解人类早期胚胎阶段的心脏发育以及再生医学中心脏分化的指导操作具有重要意义。
BackgroundCardiac differentiation from human pluripotent stem cells provides a unique opportunity to study human heart development in vitro and offers a potential cell source for cardiac regeneration. Compared to the large body of studies investigating cardiac maturation and cardiomyocyte subtype-specific induction, molecular events underlying cardiac lineage commitment from pluripotent stem cells at early stage remain poorly characterized.ResultsIn order to uncover key molecular events and regulators controlling cardiac lineage commitment from a pluripotent state during differentiation, we performed single-cell RNA-Seq sequencing and obtained high-quality data for 6879 cells collected from 6 stages during cardiac differentiation from human embryonic stem cells and identified multiple cell subpopulations with distinct molecular features. Through constructing developmental trajectory of cardiac differentiation and putative ligand-receptor interactions, we revealed crosstalk between cardiac progenitor cells and endoderm cells, which could potentially provide a cellular microenvironment supporting cardiac lineage commitment at day 5. In addition, computational analyses of single-cell RNA-Seq data unveiled ETS1 (ETS Proto-Oncogene 1) activation as an important downstream event induced by crosstalk between cardiac progenitor cells and endoderm cells. Consistent with the findings from single-cell analysis, chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-Seq) against ETS1 revealed genomic occupancy of ETS1 at cardiac structural genes at day 9 and day 14, whereas ETS1 depletion dramatically compromised cardiac differentiation.ConclusionTogether, our study not only characterized the molecular features of different cell types and identified ETS1 as a crucial factor induced by cell-cell crosstalk contributing to cardiac lineage commitment from a pluripotent state, but may also have important implications for understanding human heart development at early embryonic stage, as well as directed manipulation of cardiac differentiation in regenerative medicine.