c-JUN is a barrier in hESC to cardiomyocyte transition
Hui Zhong1,2, Ran Zhang1,3,4, Guihuan Li3
1Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, China.
Insights
Loss of c-JUN surprisingly enhances human cardiomyocyte generation by increasing chromatin accessibility and H3K4me3 deposition, revealing a novel role in heart development regulation.
Area of Science:
- Developmental Biology
- Stem Cell Biology
- Epigenetics
Background:
- c-JUN loss causes embryonic lethality in mice, linked to cardiac system defects.
- The role of c-JUN in human cardiomyocyte cell fate determination is currently unknown.
Purpose of the Study:
- To investigate the function of c-JUN in the in vitro differentiation of human pluripotent stem cells into cardiomyocytes.
- To elucidate the molecular mechanisms by which c-JUN influences human heart development.
Main Methods:
- In vitro differentiation of human pluripotent stem cells (hPSCs) into cardiomyocytes.
- Chromatin accessibility analysis using ATAC-seq.
- Histone modification profiling using ChIP-seq for H3K4me3.
- Gene expression analysis via single-cell RNA-seq.
- Pharmacological inhibition of histone demethylase activity.
Main Results:
- Knockout of c-JUN significantly improved cardiomyocyte generation, evidenced by increased TNNT2+ cells.
- c-JUN deficiency led to enhanced chromatin accessibility at key cardiac regulatory elements.
- c-JUN knockout increased RBBP5 and SETD1B expression, boosting H3K4me3 levels on cardiogenesis genes.
- Pharmacological inhibition of histone demethylase mimicked the c-JUN KO phenotype, enhancing cardiomyocyte production.
- Single-cell RNA-seq revealed that c-JUN knockout activated more cardiogenesis-related regulons.
Conclusions:
- c-JUN negatively regulates human cardiomyocyte differentiation.
- c-JUN controls cardiomyocyte cell fate by modulating H3K4me3 epigenetic marks and chromatin accessibility.
- These findings provide new insights into the role of c-JUN in human heart development.
Abstract:
Loss of c-JUN leads to early mouse embryonic death, possibly because of a failure to develop a normal cardiac system. How c-JUN regulates human cardiomyocyte cell fate remains unknown. Here, we used the in vitro differentiation of human pluripotent stem cells into cardiomyocytes to study the role of c-JUN. Surprisingly, the knockout of c-JUN improved cardiomyocyte generation, as determined by the number of TNNT2+ cells. ATAC-seq data showed that the c-JUN defect led to increased chromatin accessibility on critical regulatory elements related to cardiomyocyte development. ChIP-seq data showed that the knockout c-JUN increased RBBP5 and SETD1B expression, leading to improved H3K4me3 deposition on key genes that regulate cardiogenesis. The c-JUN KO phenotype could be copied using the histone demethylase inhibitor CPI-455, which also up-regulated H3K4me3 levels and increased cardiomyocyte generation. Single-cell RNA-seq data defined three cell branches, and knockout c-JUN activated more regulons that are related to cardiogenesis. In summary, our data demonstrated that c-JUN could regulate cardiomyocyte cell fate by modulating H3K4me3 modification and chromatin accessibility and shed light on how c-JUN regulates heart development in humans.
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