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CDH18 is a fetal epicardial biomarker regulating differentiation towards vascular smooth muscle cells

Junghof, Julia 京都大学 DOI:10.14989/doctor.k23815

2022.03.23

概要

The epicardium is a mesothelial layer that covers the myocardium and serves as a progenitor source supporting cardiac development, repair and regeneration. During development the epicardium is derived from the pro-epicardium (PE), a transient structure that spreads over the heart tube, and contributes to different cardiac lineage descendants, making it essential for cardiogenesis. Epicardial cell plasticity is governed by finely tuned signaling that regulates the epicardial-to-mesenchymal transition (EMT) and the cell-fate decisions of epicardial-derived cells (EPDCs). Interestingly, the epicardium reactivates upon cardiac injury promoting cardiac repair and regeneration. In recent years, the epicardium has emerged as a therapeutic target.

However, the biology of the epicardium as well as its repair and regeneration processes are still poorly understood as powerful tools to investigate epicardial function, including markers with pivotal roles in developmental signaling, are lacking. In this study, human induced pluripotent stem cells (hiPSCs) were used to recapitulate epicardiogenesis and successfully generate hiPSC-epicardial-like (EPI) cells with high efficiency. By a retro-perspective analysis of public available RNA-seq datasets, type II classical cadherin CDH18 was identified as a biomarker defining lineage specification in human active epicardium, and its expression in EPI cells as well as in explant cells derived from mouse embryonic hearts at E14 was demonstrated. The exclusiveness of CDH18 expression to epicardial cells and its absence in EPDCs were confirmed, thus strengthening the importance of CDH18 as a biomarker compared to other common epicardial marker genes.

The knockdown of CDH18 led to the loss of epicardial identity accompanied by the onset of EMT. The loss of CDH18 induced the activation of the Wnt signaling pathway and in combination with loss of TCF21 resulted in a cell-fate-specific differentiation towards cardiac smooth muscle cells (SMCs). This effect was found to be more apparent in cells representing a more PE-like state compared to cells representing a fetal-like epicardial stage. While the ectopic expression of CDH18 was not sufficient to inhibit the directed differentiation of SMCs, it did reduce marker expression and decreased the invasive potential of SMCs derived from hiPSCs. Furthermore, the correlation of GATA4 expression with epicardial CDH18 expression was shown, hinting at a potential regulator role. These results highlight the importance of tracing CDH18 expression in hiPSC-derived epicardial cells, providing a model for investigating epicardial function in human development and disease and enabling new possibilities for regenerative medicine.

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