Cargando…

Identification of circRNA Expression Profiles in BMSCs from Glucocorticoid-Induced Osteoporosis Model

BACKGROUND: Circular RNAs (circRNAs) contribute to the regulation of many diseases. However, little is known about the role of circRNAs in the development of glucocorticoid-induced osteoporosis (GIOP). The present study is aimed at systematically characterizing the circRNA expression profiles in GIO...

Descripción completa

Detalles Bibliográficos
Autores principales: Chen, Zhipeng, Lin, Wei, Zhao, Shengli, Mo, Xiaoyi, Wen, Zhenxing, Cheung, Wing Hoi, Fu, Dan, Chen, Bailing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8837445/
https://www.ncbi.nlm.nih.gov/pubmed/35154330
http://dx.doi.org/10.1155/2022/3249737
_version_ 1784649910257188864
author Chen, Zhipeng
Lin, Wei
Zhao, Shengli
Mo, Xiaoyi
Wen, Zhenxing
Cheung, Wing Hoi
Fu, Dan
Chen, Bailing
author_facet Chen, Zhipeng
Lin, Wei
Zhao, Shengli
Mo, Xiaoyi
Wen, Zhenxing
Cheung, Wing Hoi
Fu, Dan
Chen, Bailing
author_sort Chen, Zhipeng
collection PubMed
description BACKGROUND: Circular RNAs (circRNAs) contribute to the regulation of many diseases. However, little is known about the role of circRNAs in the development of glucocorticoid-induced osteoporosis (GIOP). The present study is aimed at systematically characterizing the circRNA expression profiles in GIOP and predict the potential functions of the associated regulatory networks. METHODS: A small animal GIOP model was developed in Sprague-Dawley rats given daily intraperitoneal doses of the synthetic glucocorticoid dexamethasone. Micro-CT and bone histomorphometry were performed to characterize the bone loss. Alizarin red S (ARS) staining and alkaline phosphatase (ALP) activity were assessed to determine the osteogenic differentiation potential of BMSCs. RNA sequencing was performed to identify differentially expressed circRNAs in BMSCs between the GIOP and normal groups, which were validated by qRT-PCR. siRNA interference experiments were used to demonstrate their function. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to predict the functions of differentially expressed circRNAs. The microRNA (miRNA) targets of the circRNAs and circRNA-miRNA interactions were predicted. RESULTS: Micro-CT and bone histomorphometry confirmed the rat GIOP model. Both ARS intensity and ALP activity were decreased in GIOP BMSCs. Seventeen circRNAs were identified by fold change = 2.0, p < 0.05, and false discovery rate < 0.05, of which 7 were upregulated and 10 were downregulated. The qRT-PCR results of the selected circRNAs were consistent with the RNA-seq results and showed that circARSB and circAKT3 were significantly upregulated, while circPTEN and circTRPM7 were downregulated in the GIOP group. Further functional experiments found that downregulation of circARSB and circPTEN expression resulted in a corresponding change in osteogenic differentiation, suggesting that circARSB negatively, while circPTEN positively, regulates BMSC osteogenic differentiation. Analysis of circRNA-targeted miRNAs predicted that miR-135a-5p was associated with circARSB and circAKT3, and miR-881-3p was associated with circPTEN and circTRPM7. Furthermore, the signalling pathways associated with these differentially expressed circRNAs were predicted. CONCLUSIONS: The present study identified circARSB, circAKT3, circPTEN, and circTRPM7 as being associated with osteogenic differentiation during GIOP through a circRNA-targeted miRNA-mRNA axis, which might provide insight into the pathophysiological mechanism of GIOP.
format Online
Article
Text
id pubmed-8837445
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Hindawi
record_format MEDLINE/PubMed
spelling pubmed-88374452022-02-12 Identification of circRNA Expression Profiles in BMSCs from Glucocorticoid-Induced Osteoporosis Model Chen, Zhipeng Lin, Wei Zhao, Shengli Mo, Xiaoyi Wen, Zhenxing Cheung, Wing Hoi Fu, Dan Chen, Bailing Stem Cells Int Research Article BACKGROUND: Circular RNAs (circRNAs) contribute to the regulation of many diseases. However, little is known about the role of circRNAs in the development of glucocorticoid-induced osteoporosis (GIOP). The present study is aimed at systematically characterizing the circRNA expression profiles in GIOP and predict the potential functions of the associated regulatory networks. METHODS: A small animal GIOP model was developed in Sprague-Dawley rats given daily intraperitoneal doses of the synthetic glucocorticoid dexamethasone. Micro-CT and bone histomorphometry were performed to characterize the bone loss. Alizarin red S (ARS) staining and alkaline phosphatase (ALP) activity were assessed to determine the osteogenic differentiation potential of BMSCs. RNA sequencing was performed to identify differentially expressed circRNAs in BMSCs between the GIOP and normal groups, which were validated by qRT-PCR. siRNA interference experiments were used to demonstrate their function. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to predict the functions of differentially expressed circRNAs. The microRNA (miRNA) targets of the circRNAs and circRNA-miRNA interactions were predicted. RESULTS: Micro-CT and bone histomorphometry confirmed the rat GIOP model. Both ARS intensity and ALP activity were decreased in GIOP BMSCs. Seventeen circRNAs were identified by fold change = 2.0, p < 0.05, and false discovery rate < 0.05, of which 7 were upregulated and 10 were downregulated. The qRT-PCR results of the selected circRNAs were consistent with the RNA-seq results and showed that circARSB and circAKT3 were significantly upregulated, while circPTEN and circTRPM7 were downregulated in the GIOP group. Further functional experiments found that downregulation of circARSB and circPTEN expression resulted in a corresponding change in osteogenic differentiation, suggesting that circARSB negatively, while circPTEN positively, regulates BMSC osteogenic differentiation. Analysis of circRNA-targeted miRNAs predicted that miR-135a-5p was associated with circARSB and circAKT3, and miR-881-3p was associated with circPTEN and circTRPM7. Furthermore, the signalling pathways associated with these differentially expressed circRNAs were predicted. CONCLUSIONS: The present study identified circARSB, circAKT3, circPTEN, and circTRPM7 as being associated with osteogenic differentiation during GIOP through a circRNA-targeted miRNA-mRNA axis, which might provide insight into the pathophysiological mechanism of GIOP. Hindawi 2022-02-04 /pmc/articles/PMC8837445/ /pubmed/35154330 http://dx.doi.org/10.1155/2022/3249737 Text en Copyright © 2022 Zhipeng Chen et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Chen, Zhipeng
Lin, Wei
Zhao, Shengli
Mo, Xiaoyi
Wen, Zhenxing
Cheung, Wing Hoi
Fu, Dan
Chen, Bailing
Identification of circRNA Expression Profiles in BMSCs from Glucocorticoid-Induced Osteoporosis Model
title Identification of circRNA Expression Profiles in BMSCs from Glucocorticoid-Induced Osteoporosis Model
title_full Identification of circRNA Expression Profiles in BMSCs from Glucocorticoid-Induced Osteoporosis Model
title_fullStr Identification of circRNA Expression Profiles in BMSCs from Glucocorticoid-Induced Osteoporosis Model
title_full_unstemmed Identification of circRNA Expression Profiles in BMSCs from Glucocorticoid-Induced Osteoporosis Model
title_short Identification of circRNA Expression Profiles in BMSCs from Glucocorticoid-Induced Osteoporosis Model
title_sort identification of circrna expression profiles in bmscs from glucocorticoid-induced osteoporosis model
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8837445/
https://www.ncbi.nlm.nih.gov/pubmed/35154330
http://dx.doi.org/10.1155/2022/3249737
work_keys_str_mv AT chenzhipeng identificationofcircrnaexpressionprofilesinbmscsfromglucocorticoidinducedosteoporosismodel
AT linwei identificationofcircrnaexpressionprofilesinbmscsfromglucocorticoidinducedosteoporosismodel
AT zhaoshengli identificationofcircrnaexpressionprofilesinbmscsfromglucocorticoidinducedosteoporosismodel
AT moxiaoyi identificationofcircrnaexpressionprofilesinbmscsfromglucocorticoidinducedosteoporosismodel
AT wenzhenxing identificationofcircrnaexpressionprofilesinbmscsfromglucocorticoidinducedosteoporosismodel
AT cheungwinghoi identificationofcircrnaexpressionprofilesinbmscsfromglucocorticoidinducedosteoporosismodel
AT fudan identificationofcircrnaexpressionprofilesinbmscsfromglucocorticoidinducedosteoporosismodel
AT chenbailing identificationofcircrnaexpressionprofilesinbmscsfromglucocorticoidinducedosteoporosismodel