Profiling of transcriptional and epigenetic changes during directed endothelial differentiation of human embryonic stem cells identifies FOXA2 as a marker of early mesoderm commitment

Howard, L., MacKenzie, R. M., Pchelintsev, N. A., McBryan, T., McClure, J. D. , McBride, M. W. , Kane, N. M., Adams, P. D., Milligan, G. and Baker, A. H. (2013) Profiling of transcriptional and epigenetic changes during directed endothelial differentiation of human embryonic stem cells identifies FOXA2 as a marker of early mesoderm commitment. Stem Cell Research and Therapy, 4(2), Art. 36. (doi:10.1186/scrt192)

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Publisher's URL: http://dx.doi.org/10.1186/scrt192

Abstract

Introduction: Differentiation of vascular endothelial cells (ECs) in clinically relevant numbers for injection into ischaemic areas could offer therapeutic potential in the treatment of cardiovascular conditions, including myocardial infarction, peripheral vascular disease and stroke. While we and others have demonstrated successful generation of functional endothelial-like cells from human embryonic stem cells (hESCs), little is understood regarding the complex transcriptional and epigenetic changes that occur during differentiation, in particular during early commitment to a mesodermal lineage. Methods: We performed the first gene expression microarray study of hESCs undergoing directed differentiation to ECs using a monolayer-based, feeder-free and serum-free protocol. Microarray results were confirmed by quantitative RT-PCR and immunocytochemistry, and chromatin immunoprecipitation (ChIP)-PCR analysis was utilised to determine the bivalent status of differentially expressed genes. Results: We identified 22 transcription factors specific to early mesoderm commitment. Among these factors, FOXA2 was observed to be the most significantly differentially expressed at the hESC–EC day 2 timepoint. ChIP-PCR analysis revealed that the FOXA2 transcription start site is bivalently marked with histone modifications for both gene activation (H3K4me3) and repression (H3K27me3) in hESCs, suggesting the transcription factor may be a key regulator of hESC differentiation. Conclusion: This enhanced knowledge of the lineage commitment process will help improve the design of directed differentiation protocols, increasing the yield of endothelial-like cells for regenerative medicine therapies in cardiovascular disease.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Baker, Professor Andrew and Adams, Professor Peter and Pchelintsev, Dr Nikolay and Kane, Dr Nicole and McBride, Dr Martin and Milligan, Professor Graeme and MacKenzie, Dr Ruth and Howard, Miss Lynsey and McBryan, Dr Anthony and McClure, Dr John
Authors: Howard, L., MacKenzie, R. M., Pchelintsev, N. A., McBryan, T., McClure, J. D., McBride, M. W., Kane, N. M., Adams, P. D., Milligan, G., and Baker, A. H.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cancer Sciences
College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Stem Cell Research and Therapy
Publisher:BioMed Central
ISSN:1757-6512
Copyright Holders:Copyright © 2013 The Authors
First Published:First published in Stem Cell Research and Therapy 4(2):36
Publisher Policy:Reproduced under a Creative Commons License
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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
531341In vitro and in vivo analysis of stem cell commitment to vascular endothelial cellsAndrew BakerBritish Heart Foundation (BHF)SP/10/005/28298RI CARDIOVASCULAR & MEDICAL SCIENCES
583361BHF Chair of Translational Cardiovascular SciencesAndrew BakerBritish Heart Foundation (BHF)CH/11/2/28733RI CARDIOVASCULAR & MEDICAL SCIENCES