RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification
Abstract 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 is...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-05-01
|
| Series: | BMC Nephrology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12882-022-02823-6 |
| _version_ | 1811255040592052224 |
|---|---|
| author | Zhong Peng Yingjie Duan Shuzhu Zhong Juan Chen Jianlong Li Zhangxiu He |
| author_facet | Zhong Peng Yingjie Duan Shuzhu Zhong Juan Chen Jianlong Li Zhangxiu He |
| author_sort | Zhong Peng |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-04-12T17:17:53Z |
| format | Article |
| id | doaj.art-7ea4533c48be45c29a557fdf3250de68 |
| institution | Directory Open Access Journal |
| issn | 1471-2369 |
| language | English |
| last_indexed | 2024-04-12T17:17:53Z |
| publishDate | 2022-05-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Nephrology |
| spelling | doaj.art-7ea4533c48be45c29a557fdf3250de682022-12-22T03:23:35ZengBMCBMC Nephrology1471-23692022-05-0123111110.1186/s12882-022-02823-6RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcificationZhong Peng0Yingjie Duan1Shuzhu Zhong2Juan Chen3Jianlong Li4Zhangxiu He5The First Affiliated Hospital, Department of Gastroenterology, Hengyang Medical School, University of South ChinaThe First Affiliated Hospital, Department of Nephrology, Hengyang Medical School, University of South ChinaThe First Affiliated Hospital, Department of Nephrology, Hengyang Medical School, University of South ChinaSchool of Food and Biological Engineering, Hefei University of TechnologyDepartment of Orthopedic Surgery, Nanfang Hospital, Southern Medical UniversityThe First Affiliated Hospital, Department of Nephrology, Hengyang Medical School, University of South ChinaAbstract 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.https://doi.org/10.1186/s12882-022-02823-6HyperphosphatemiaExtracellular vesicles (EVs)Vascular calcificationHigh-throughput sequencing (HTS) |
| spellingShingle | Zhong Peng Yingjie Duan Shuzhu Zhong Juan Chen Jianlong Li Zhangxiu He RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification BMC Nephrology Hyperphosphatemia Extracellular vesicles (EVs) Vascular calcification High-throughput sequencing (HTS) |
| 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 | Hyperphosphatemia Extracellular vesicles (EVs) Vascular calcification High-throughput sequencing (HTS) |
| url | https://doi.org/10.1186/s12882-022-02823-6 |
| work_keys_str_mv | AT zhongpeng rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification AT yingjieduan rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification AT shuzhuzhong rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification AT juanchen rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification AT jianlongli rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification AT zhangxiuhe rnaseqanalysisofextracellularvesiclesfromhyperphosphatemiastimulatedendothelialcellsprovidesinsightintothemechanismunderlyingvascularcalcification |