Hyperoxia impairs pro-angiogenic RNA production in preterm endothelial colony-forming cells

Megan A. Ahern; Claudine P. Black; Gregory J. Seedorf; Christopher D. Baker; Douglas P. Shepherd; Megan A. Ahern; Claudine P. Black; Gregory J. Seedorf; Christopher D. Baker; Douglas P. Shepherd

doi:10.3934/biophy.2017.2.284

AIMS Biophysics

2017, Volume 4, Issue 2: 284-297. doi: 10.3934/biophy.2017.2.284

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Hyperoxia impairs pro-angiogenic RNA production in preterm endothelial colony-forming cells

1.
Department of Physics, University of Colorado Denver, Denver, CO 80212, USA
2.
Pediatric Heart Lung Center, Department of Pediatrics, University of Colorado Denver Anschutz Medical Campus, Aurora CO 80045, USA

Received: 09 January 2017 Accepted: 10 April 2017 Published: 19 April 2017

Disruptions in the response of endothelial progenitor cells to changes in oxygen environment may present a possible mechanism behind multiple pediatric pulmonary disease models, such as bronchopulmonary dysplasia. Using high-throughput fixed single-cell protein and RNA imaging, we have created “stop-motion” movies of Thymosin β4 (Tβ4) and Hypoxia Inducible Factor 1α (HIF-1α) protein expression and vascular endothelial growth factor (vegf) and endothelial nitric oxide synthase (eNOS) mRNA in human umbilical cord-derived endothelial colony-forming cells (ECFC). ECFC were grown in vitro under both room air and hyperoxia (50% O₂). We find elevated basal Tβ4 protein expression in ECFC derived from prematurely born infants versus full term infants. Tβ4 is a potent growth hormone that additionally acts as an actin sequestration protein and regulates the stability of HIF-1α. This basal level increase of Tβ4 is associated with lower HIF-1α nuclear localization in preterm versus term ECFC upon exposure to hyperoxia. We find altered expression in the pro-angiogenic genes vegf and eNOS, two genes that HIF-1α acts as a transcription factor for. This provides a potential link between a developmentally regulated protein and previously observed impaired function of preterm ECFC in response to hyperoxia.
- single-molecule,
- single-cell,
- fluorescence microscopy,
- endothelial colony-forming cells,
- endothelial cell biology
Citation: Megan A. Ahern, Claudine P. Black, Gregory J. Seedorf, Christopher D. Baker, Douglas P. Shepherd. Hyperoxia impairs pro-angiogenic RNA production in preterm endothelial colony-forming cells[J]. AIMS Biophysics, 2017, 4(2): 284-297. doi: 10.3934/biophy.2017.2.284

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Abstract

Disruptions in the response of endothelial progenitor cells to changes in oxygen environment may present a possible mechanism behind multiple pediatric pulmonary disease models, such as bronchopulmonary dysplasia. Using high-throughput fixed single-cell protein and RNA imaging, we have created “stop-motion” movies of Thymosin β4 (Tβ4) and Hypoxia Inducible Factor 1α (HIF-1α) protein expression and vascular endothelial growth factor (vegf) and endothelial nitric oxide synthase (eNOS) mRNA in human umbilical cord-derived endothelial colony-forming cells (ECFC). ECFC were grown in vitro under both room air and hyperoxia (50% O₂). We find elevated basal Tβ4 protein expression in ECFC derived from prematurely born infants versus full term infants. Tβ4 is a potent growth hormone that additionally acts as an actin sequestration protein and regulates the stability of HIF-1α. This basal level increase of Tβ4 is associated with lower HIF-1α nuclear localization in preterm versus term ECFC upon exposure to hyperoxia. We find altered expression in the pro-angiogenic genes vegf and eNOS, two genes that HIF-1α acts as a transcription factor for. This provides a potential link between a developmentally regulated protein and previously observed impaired function of preterm ECFC in response to hyperoxia.

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