BNC1 regulates cell heterogeneity in human pluripotent stem cell-derived epicardium.

Laure Gambardella,Victoria Moignard,Simon Andrews,Agathe Delaune,Paul R Riley,Berthold Göttgens,Sophie A Mcmanus,Sanjay Sinha,Nicolas Gambardella Le Novère,Maura A Morrison,Derya Sebukhan,William G Bernard

doi:10.1242/dev.174441

Laure Gambardella, Victoria Moignard + Show 10 more

Open Access

https://doi.org/10.1242/dev.174441

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Abstract

ABSTRACTThe murine developing epicardium heterogeneously expresses the transcription factors TCF21 and WT1. Here, we show that this cell heterogeneity is conserved in human epicardium, regulated by BNC1 and associated with cell fate and function. Single cell RNA sequencing of epicardium derived from human pluripotent stem cells (hPSC-epi) revealed that distinct epicardial subpopulations are defined by high levels of expression for the transcription factors BNC1 or TCF21. WT1+ cells are included in the BNC1+ population, which was confirmed in human foetal hearts. THY1 emerged as a membrane marker of the TCF21 population. We show that THY1+ cells can differentiate into cardiac fibroblasts (CFs) and smooth muscle cells (SMCs), whereas THY1− cells were predominantly restricted to SMCs. Knocking down BNC1 during the establishment of the epicardial populations resulted in a homogeneous, predominantly TCF21high population. Network inference methods using transcriptomic data from the different cell lineages derived from the hPSC-epi delivered a core transcriptional network organised around WT1, TCF21 and BNC1. This study unveils a list of epicardial regulators and is a step towards engineering subpopulations of epicardial cells with selective biological activities.

Highlights

The epicardium is an epithelium covering the heart, which is essential for normal cardiac development
Because both antibodies suitable for the detection of transcription factor 21 (TCF21) and Wilms’ tumor 1 (WT1) in human cells were rabbit in origin, we were previously limited to a flow cytometry strategy in which the presence of double-positive cells in the hPSC-epi was indirectly estimated (Iyer et al, 2015)
We identified the transcription factor BNC1 and the membrane protein THY1 as markers of the TCF21low and TCF21high populations, respectively

Summary

Introduction

The epicardium is an epithelium covering the heart, which is essential for normal cardiac development. Epicardial cells originate from the pro-epicardial (PE) organ, an outgrowth of non-cardiac, coelomic and proliferative cells located between the heart and the liver (Smits et al, 2018). The epicardium provides signals for proliferation, survival and maturation to the cardiomyocytes. The myocardium provides signals inducing proliferation and epithelial-to-mesenchymal transition. L.G., 0000-0001-5771-1565; V.M., 0000-0001-5089-5875; D.S., 0000-00027798-7953; A.D., 0000-0003-4552-8266; N.G.L.N., 0000-0002-6309-7327; S.S., 0000-0001-5900-1209 The mesenchymal cells derived from the epicardium (EPDCs) invade the myocardium and become mainly cardiac fibroblasts (CFs) and coronary smooth muscle cells (cSMCs). Surgical ablation of the PE organ leads to cardiac developmental abnormalities (Gittenberger-De Groot et al, 2000)

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