Single-cell reconstruction of differentiation trajectory reveals a critical role of ETS1 in human cardiac lineage
Hang Ruan1,2, Yingnan Liao1, Zongna Ren1
1State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, 167 North Lishi Road, Beijing, 100037, People's Republic of China.
Insights
Researchers identified ETS1 activation as key to early cardiac lineage commitment from pluripotent stem cells. This discovery sheds light on human heart development and cardiac regeneration strategies.
Area of Science:
- Stem cell biology
- Developmental biology
- Cardiovascular research
Background:
- Human pluripotent stem cells (hPSCs) offer a model for studying human heart development and potential for cardiac regeneration.
- Early molecular events driving cardiac lineage commitment from hPSCs are not well understood.
- Existing research focuses more on later cardiac maturation and specific cardiomyocyte subtypes.
Purpose of the Study:
- To identify key molecular events and regulators governing cardiac lineage commitment from a pluripotent state.
- To characterize cell subpopulations and their interactions during early cardiac differentiation.
- To investigate the role of ETS1 in cardiac lineage commitment.
Main Methods:
- Single-cell RNA sequencing (scRNA-Seq) of 6879 cells across 6 differentiation stages.
- Construction of developmental trajectories and analysis of ligand-receptor interactions.
- Chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-Seq) for ETS1.
- ETS1 depletion experiments to assess its functional role.
Main Results:
- Identification of multiple cell subpopulations with distinct molecular profiles during differentiation.
- Revealed crosstalk between cardiac progenitor cells and endoderm cells at day 5, potentially supporting lineage commitment.
- Identified ETS1 activation as a downstream event of this crosstalk.
- Demonstrated ETS1 genomic occupancy at cardiac structural genes and its essential role in cardiac differentiation.
Conclusions:
- Characterized molecular features of cell types during early cardiac differentiation.
- Identified ETS1 as a crucial factor, induced by cell-cell crosstalk, for cardiac lineage commitment.
- Findings have implications for understanding early human heart development and advancing cardiac regenerative medicine.
Background:
Cardiac 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.
Results:
In 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.
Conclusion:
Together, 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.
Related Concept Videos
01:21Lineage Commitment
00:57Cellular Differentiation
A zygote is a...
01:51Determination