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Cyclic Strain Promotes H19 Expression and Vascular Tube Formation in iPSC-Derived Endothelial Cells

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Abstract

Introduction

Induced pluripotent stem cell (iPSC)-derived endothelial cells (ECs) have the potential for therapeutic application in several cardiovascular diseases. Mechanical strain is known to regulate EC behavior and stem cell differentiation and may play a role in directing EC differentiation of iPSCs. H19, a long non-coding RNA (lncRNA), is known to affect ECs in several mechanically relevant pathologies and may play a role in this process as well. Therefore, we investigated expression changes of H19 resulting from mechanical stimulation during EC differentiation, as well as functional effects on EC tube formation.

Methods

iPSCs were subjected to 5% cyclic mechanical strain during EC differentiation. RT-PCR and flow cytometry were used to assess changes in mesoderm differentiation and gene expression in the final ECs as a result of strain. Functional outcomes of mechanically differentiated ECs were assessed with a tube formation assay and changes in H19. H19 was also overexpressed in human umbilical vein endothelial cells (HUVECs) to assess its role in non-H19-expressing ECs.

Results

Mechanical strain promoted mesoderm differentiation, marked by increased expression of brachyury 24 h after initiation of differentiation. Strain also increased expression of H19, CD31, VE-cadherin, and VEGFR2 in differentiated ECs. Strain-differentiated ECs formed tube networks with higher junction and endpoint density than statically-differentiated ECs. Overexpression of H19 in HUVECs resulted in similar patterns of tube formation.

Conclusions

H19 expression is increased by mechanical strain and promotes tube branching in iPSC-derived ECs.

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Funding

This work was supported by the National Heart, Lung, and Blood Institute (HL135790 and HL007411) and the Fondation Leducq.

Conflict of interest

No conflict of interest, financial or otherwise, are declared by Dr. Vander Roest or Dr. Merryman.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors

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Authors and Affiliations

  1. Biomedical Engineering, Vanderbilt University, Room 9445D MRB4, 2213 Garland Ave, Nashville, TN, 37212, USA

    Mark J. Vander Roest & W. David Merryman

Authors
  1. Mark J. Vander Roest
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  2. W. David Merryman
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Correspondence to W. David Merryman.

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Associate Editor Michael R. King oversaw the review of this article.

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Vander Roest, M.J., Merryman, W.D. Cyclic Strain Promotes H19 Expression and Vascular Tube Formation in iPSC-Derived Endothelial Cells. Cel. Mol. Bioeng. 13, 369–377 (2020). https://doi.org/10.1007/s12195-020-00617-0

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  • DOI: https://doi.org/10.1007/s12195-020-00617-0

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