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eNOS expression and NO release during hypoxia is inhibited by miR-200b in human endothelial cells
The nitric oxide (NO) secreted by vascular endothelium is required for the maintenance of cardiovascular homeostasis. Diminished release of NO generated by endothelial NO synthase contributes to endothelial dysfunction. Hypoxia and ischemia reduce endothelial eNOS expression via posttranscriptional...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Netherlands
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6208887/ https://www.ncbi.nlm.nih.gov/pubmed/29737439 http://dx.doi.org/10.1007/s10456-018-9620-y |
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author | Janaszak-Jasiecka, Anna Siekierzycka, Anna Bartoszewska, Sylwia Serocki, Marcin Dobrucki, Lawrence W. Collawn, James F. Kalinowski, Leszek Bartoszewski, Rafal |
author_facet | Janaszak-Jasiecka, Anna Siekierzycka, Anna Bartoszewska, Sylwia Serocki, Marcin Dobrucki, Lawrence W. Collawn, James F. Kalinowski, Leszek Bartoszewski, Rafal |
author_sort | Janaszak-Jasiecka, Anna |
collection | PubMed |
description | The nitric oxide (NO) secreted by vascular endothelium is required for the maintenance of cardiovascular homeostasis. Diminished release of NO generated by endothelial NO synthase contributes to endothelial dysfunction. Hypoxia and ischemia reduce endothelial eNOS expression via posttranscriptional mechanisms that result in NOS3 transcript destabilization. Here, we examine whether microRNAs contribute to this mechanism. We followed the kinetics of hypoxia-induced changes in NOS3 mRNA and eNOS protein levels in primary human umbilical vein endothelial cells (HUVECs). Utilizing in silico predictive protocols to identify potential miRNAs that regulate eNOS expression, we identified miR-200b as a candidate. We established the functional miR-200b target sequence within the NOS3 3′UTR, and demonstrated that manipulation of the miRNA levels during hypoxia using miR-200b mimics and antagomirs regulates eNOS levels, and established that miR-200b physiologically limits eNOS expression during hypoxia. Furthermore, we demonstrated that the specific ablation of the hypoxic induction of miR-200b in HUVECs restored eNOS-driven hypoxic NO release to the normoxic levels. To determine whether miR-200b might be the only miRNA that had this effect, we utilized Next Generation Sequencing (NGS) to follow hypoxia-induced changes in the miRNA levels in HUVECS and found 83 novel hypoxamiRs, with two candidate miRNAs besides miR-200b that could potentially influence eNOS levels. Taken together, the data establish miR-200b-eNOS regulation as a first hypoxamiR-based mechanism that limits NO bioavailability during hypoxia in endothelial cells, and show that hypoxamiRs could become useful therapeutic targets for cardiovascular diseases and other hypoxic-related diseases including various types of cancer. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10456-018-9620-y) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6208887 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Springer Netherlands |
record_format | MEDLINE/PubMed |
spelling | pubmed-62088872018-11-09 eNOS expression and NO release during hypoxia is inhibited by miR-200b in human endothelial cells Janaszak-Jasiecka, Anna Siekierzycka, Anna Bartoszewska, Sylwia Serocki, Marcin Dobrucki, Lawrence W. Collawn, James F. Kalinowski, Leszek Bartoszewski, Rafal Angiogenesis Brief Communication The nitric oxide (NO) secreted by vascular endothelium is required for the maintenance of cardiovascular homeostasis. Diminished release of NO generated by endothelial NO synthase contributes to endothelial dysfunction. Hypoxia and ischemia reduce endothelial eNOS expression via posttranscriptional mechanisms that result in NOS3 transcript destabilization. Here, we examine whether microRNAs contribute to this mechanism. We followed the kinetics of hypoxia-induced changes in NOS3 mRNA and eNOS protein levels in primary human umbilical vein endothelial cells (HUVECs). Utilizing in silico predictive protocols to identify potential miRNAs that regulate eNOS expression, we identified miR-200b as a candidate. We established the functional miR-200b target sequence within the NOS3 3′UTR, and demonstrated that manipulation of the miRNA levels during hypoxia using miR-200b mimics and antagomirs regulates eNOS levels, and established that miR-200b physiologically limits eNOS expression during hypoxia. Furthermore, we demonstrated that the specific ablation of the hypoxic induction of miR-200b in HUVECs restored eNOS-driven hypoxic NO release to the normoxic levels. To determine whether miR-200b might be the only miRNA that had this effect, we utilized Next Generation Sequencing (NGS) to follow hypoxia-induced changes in the miRNA levels in HUVECS and found 83 novel hypoxamiRs, with two candidate miRNAs besides miR-200b that could potentially influence eNOS levels. Taken together, the data establish miR-200b-eNOS regulation as a first hypoxamiR-based mechanism that limits NO bioavailability during hypoxia in endothelial cells, and show that hypoxamiRs could become useful therapeutic targets for cardiovascular diseases and other hypoxic-related diseases including various types of cancer. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10456-018-9620-y) contains supplementary material, which is available to authorized users. Springer Netherlands 2018-05-08 2018 /pmc/articles/PMC6208887/ /pubmed/29737439 http://dx.doi.org/10.1007/s10456-018-9620-y Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Brief Communication Janaszak-Jasiecka, Anna Siekierzycka, Anna Bartoszewska, Sylwia Serocki, Marcin Dobrucki, Lawrence W. Collawn, James F. Kalinowski, Leszek Bartoszewski, Rafal eNOS expression and NO release during hypoxia is inhibited by miR-200b in human endothelial cells |
title | eNOS expression and NO release during hypoxia is inhibited by miR-200b in human endothelial cells |
title_full | eNOS expression and NO release during hypoxia is inhibited by miR-200b in human endothelial cells |
title_fullStr | eNOS expression and NO release during hypoxia is inhibited by miR-200b in human endothelial cells |
title_full_unstemmed | eNOS expression and NO release during hypoxia is inhibited by miR-200b in human endothelial cells |
title_short | eNOS expression and NO release during hypoxia is inhibited by miR-200b in human endothelial cells |
title_sort | enos expression and no release during hypoxia is inhibited by mir-200b in human endothelial cells |
topic | Brief Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6208887/ https://www.ncbi.nlm.nih.gov/pubmed/29737439 http://dx.doi.org/10.1007/s10456-018-9620-y |
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