RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification

BACKGROUND: Hyperphosphatemia (HP) is associated with vascular calcification (VC) in chronic kidney disease (CKD). However, relationship between HP-induced-endothelial extracellular vesicles (HP-EC-EVs) and VC is unclear, and miR expression in HP-EC-EVs has not been determined. METHODS: We isolated...

Descripción completa

Detalles Bibliográficos
Autores principales: Peng, Zhong, Duan, Yingjie, Zhong, Shuzhu, Chen, Juan, Li, Jianlong, He, Zhangxiu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Research
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9123672/
https://www.ncbi.nlm.nih.gov/pubmed/35597927
http://dx.doi.org/10.1186/s12882-022-02823-6
_version_ 1784711600135995392
author Peng, Zhong
Duan, Yingjie
Zhong, Shuzhu
Chen, Juan
Li, Jianlong
He, Zhangxiu
author_facet Peng, Zhong
Duan, Yingjie
Zhong, Shuzhu
Chen, Juan
Li, Jianlong
He, Zhangxiu
author_sort Peng, Zhong
collection PubMed
description BACKGROUND: Hyperphosphatemia (HP) is associated with vascular calcification (VC) in chronic kidney disease (CKD). However, relationship between HP-induced-endothelial extracellular vesicles (HP-EC-EVs) and VC is unclear, and miR expression in HP-EC-EVs has not been determined. METHODS: We isolated HP-EC-EVs from endothelial cells with HP and observed that HP-EC-EVs were up-taken by vascular smooth muscle cells (VSMCs). HP-EC-EVs inducing calcium deposition was characterized by Alizarin Red S, colourimetric analysis and ALP activity. To investigate the mechanism of HP-EC-EVs-induced VSMC calcification, RNA-sequencing for HP-EC-EVs was performed. RESULTS: We first demonstrated that HP-EC-EVs induced VSMC calcification in vitro. RNA-seq analysis of HP-EC-EVs illustrated that one known miR (hsa-miR-3182) was statistically up-regulated and twelve miRs were significantly down-regulated, which was verified by qRT-PCR. We predicted 58,209 and 74,469 target genes for those down- and up-regulated miRs respectively through miRDB, miRWalk and miRanda databases. GO terms showed that down- and up-regulated targets were mostly enriched in calcium-dependent cell–cell adhesion via plama membrane cell-adhesion molecules (GO:0,016,338, BP) and cell adhesion (GO:0,007,155, BP), plasma membrane (GO:0,005,886, CC), and metal ion binding (GO:0,046,914, MF) and ATP binding (GO:0,005,524, MF) respectively. Top-20 pathways by KEGG analysis included calcium signaling pathway, cAMP signaling pathway, and ABC transporters, which were closely related to VC. CONCLUSION: Our results indicated that those significantly altered miRs, which were packaged in HP-EC-EVs, may play an important role in VC by regulating related pathways. It may provide novel insight into the mechanism of CKD calcification. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12882-022-02823-6.
format Online
Article
Text
id pubmed-9123672
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-91236722022-05-22 RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification Peng, Zhong Duan, Yingjie Zhong, Shuzhu Chen, Juan Li, Jianlong He, Zhangxiu BMC Nephrol Research BACKGROUND: Hyperphosphatemia (HP) is associated with vascular calcification (VC) in chronic kidney disease (CKD). However, relationship between HP-induced-endothelial extracellular vesicles (HP-EC-EVs) and VC is unclear, and miR expression in HP-EC-EVs has not been determined. METHODS: We isolated HP-EC-EVs from endothelial cells with HP and observed that HP-EC-EVs were up-taken by vascular smooth muscle cells (VSMCs). HP-EC-EVs inducing calcium deposition was characterized by Alizarin Red S, colourimetric analysis and ALP activity. To investigate the mechanism of HP-EC-EVs-induced VSMC calcification, RNA-sequencing for HP-EC-EVs was performed. RESULTS: We first demonstrated that HP-EC-EVs induced VSMC calcification in vitro. RNA-seq analysis of HP-EC-EVs illustrated that one known miR (hsa-miR-3182) was statistically up-regulated and twelve miRs were significantly down-regulated, which was verified by qRT-PCR. We predicted 58,209 and 74,469 target genes for those down- and up-regulated miRs respectively through miRDB, miRWalk and miRanda databases. GO terms showed that down- and up-regulated targets were mostly enriched in calcium-dependent cell–cell adhesion via plama membrane cell-adhesion molecules (GO:0,016,338, BP) and cell adhesion (GO:0,007,155, BP), plasma membrane (GO:0,005,886, CC), and metal ion binding (GO:0,046,914, MF) and ATP binding (GO:0,005,524, MF) respectively. Top-20 pathways by KEGG analysis included calcium signaling pathway, cAMP signaling pathway, and ABC transporters, which were closely related to VC. CONCLUSION: Our results indicated that those significantly altered miRs, which were packaged in HP-EC-EVs, may play an important role in VC by regulating related pathways. It may provide novel insight into the mechanism of CKD calcification. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12882-022-02823-6. BioMed Central 2022-05-21 /pmc/articles/PMC9123672/ /pubmed/35597927 http://dx.doi.org/10.1186/s12882-022-02823-6 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Peng, Zhong
Duan, Yingjie
Zhong, Shuzhu
Chen, Juan
Li, Jianlong
He, Zhangxiu
RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification
title RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification
title_full RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification
title_fullStr RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification
title_full_unstemmed RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification
title_short RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification
title_sort rna-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9123672/
https://www.ncbi.nlm.nih.gov/pubmed/35597927
http://dx.doi.org/10.1186/s12882-022-02823-6
work_keys_str_mv AT pengzhong rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification
AT duanyingjie rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification
AT zhongshuzhu rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification
AT chenjuan rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification
AT lijianlong rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification
AT hezhangxiu rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification

Ejemplares similares