STAT3 mediates differentiation and maintenance of human pluripotent stem-derived endothelial cells

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Title: STAT3 mediates differentiation and maintenance of human pluripotent stem-derived endothelial cells
Author(s): Foldes, G
Lawlor, K
Harding, SE
Randi, AM
Item Type: Conference Paper
Abstract: Background: High plasticity derivatives of human pluripotent stem cells (hPSC) such as embryonic stem cells (hESC) are being intensively developed for their use in endothelial replacement. Methods/Results: In this study, we found that transient addition of Activin A, followed by culture with VEGF165, BMP4 and FGF2 was an effective mechanism to induce differentiation of hESC toward the endothelial lineage. Indeed, human GeneChip microarray analysis revealed that endothelial gene regulatory networks were gradually increased during 12 days of differentiation. Isolated hESC-derived endothelial cells (hESC-EC) expressed mature endothelial-associated genes, including CD31, NRP1, VE-cadherin, Tie2, VWF and ICAM2, reaching levels comparable with human umbilical cord vascular endothelial cells by day 19. We found that a network of CD31+ tubes comprising endothelial precursor cells had formed in culture from 10 days after start of differentiation of hESC. As assessed by automated high content microscopy, the alignment and tube formation of the newly formed CD31+ vascular network were markedly decreased in response to C188-9, a novel small molecule inhibitor of STAT3 transcription factor (tube length: 57%, connected tube area: 22% of those in control, both p<0.001). Human ESC-EC were capable of transdifferentiating into mesenchymal cells in long-term cultures. Endothelial-mesenchymal transition was characterised by gradual loss of endothelial marker expression and increased mesenchymal marker FSP1 expression. We found that inhibition of STAT3 tyr705 phosphorylation by C188-9 resulted in a decreased proliferation of FSP1+ mesenchymal cells (2-fold decrease in Ki67%-positive population, p<0.001), and subsequently reduced number of FSP+ cells (36% reduction, p<0.05). At the same time, C188-9 increased the number of CD31+ hESC-EC by 30% (p=0.05, n=6). Viability remained unchanged in C188-9-treated cells (Topro3 necrosis marker, p=0.32, n=3). Conclusions: These results suggest that STAT3 pathway may play a dual role in the generation and maintenance of hPSC-derived endothelial cells. Elucidation of molecular signals during endothelial differentiation of hPSC may allow specific targeting of their activities to enhance differentiation and promote vascular tissue regeneration.
Publication Date: 23-Apr-2018
Date of Acceptance: 12-Mar-2018
URI: http://hdl.handle.net/10044/1/60143
DOI: https://dx.doi.org/10.1093/cvr/cvy060.108
ISSN: 0008-6363
Publisher: OXFORD UNIV PRESS
Start Page: S39
End Page: S39
Journal / Book Title: CARDIOVASCULAR RESEARCH
Volume: 114
Issue: suppl_1
Copyright Statement: © 2018 The Author. Published on behalf of the European Society of Cardiology. All rights reserved. For permissions please email: Journals.permissions@oup.com. This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/about_us/legal/notices). This is a pre-copy-editing, author-produced version of an article accepted for publication in Cardiovascular Research following peer review. The definitive publisher-authenticated version is available online at: https://dx.doi.org/10.1093/cvr/cvy060.108
Sponsor/Funder: British Heart Foundation
Funder's Grant Number: RE/13/4/30184
Conference Name: 5th Congress of the ESC-Council-on-Basic-Cardiovascular-Science on Frontiers in Cardio Vascular Biology
Keywords: Science & Technology
Life Sciences & Biomedicine
Cardiac & Cardiovascular Systems
Cardiovascular System & Cardiology
Science & Technology
Life Sciences & Biomedicine
Cardiac & Cardiovascular Systems
Cardiovascular System & Cardiology
1102 Cardiovascular Medicine And Haematology
Cardiovascular System & Hematology
Publication Status: Published
Start Date: 2018-04-20
Finish Date: 2018-04-22
Conference Place: Vienna, AUSTRIA
Embargo Date: 2019-04-23
Online Publication Date: 2018-04-23
Appears in Collections:National Heart and Lung Institute
Faculty of Medicine



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