circRNAome profiling reveals circFgfr2 regulates myogenesis and muscle regeneration via a feedback loop
Abstract Background Circular RNAs (circRNAs) represent a novel class of non‐coding RNAs formed by a covalently closed loop and play crucial roles in many biological processes. Several circRNAs associated with myogenesis have been reported. However, the dynamic expression, function, and mechanism of...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2022-02-01
|
| Series: | Journal of Cachexia, Sarcopenia and Muscle |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jcsm.12859 |
| _version_ | 1797826745483657216 |
|---|---|
| author | Junyu Yan Yalan Yang Xinhao Fan Guoming Liang Zishuai Wang Jiju Li Liyuan Wang Yun Chen Adeyinka Abiola Adetula Yijie Tang Kui Li Dazhi Wang Zhonglin Tang |
| author_facet | Junyu Yan Yalan Yang Xinhao Fan Guoming Liang Zishuai Wang Jiju Li Liyuan Wang Yun Chen Adeyinka Abiola Adetula Yijie Tang Kui Li Dazhi Wang Zhonglin Tang |
| author_sort | Junyu Yan |
| collection | DOAJ |
| description | Abstract Background Circular RNAs (circRNAs) represent a novel class of non‐coding RNAs formed by a covalently closed loop and play crucial roles in many biological processes. Several circRNAs associated with myogenesis have been reported. However, the dynamic expression, function, and mechanism of circRNAs during myogenesis and skeletal muscle development are largely unknown. Methods Strand‐specific RNA‐sequencing (RNA‐seq) and microarray datasets were used to profile the dynamic circRNAome landscape during skeletal muscle development and myogenic differentiation. Bioinformatics analyses were used to characterize the circRNAome and identify candidate circRNAs associated with myogenesis. Bulk and single‐cell RNA‐seq were performed to identify the downstream genes and pathways of circFgfr2. The primary myoblast cells, C2C12 cells, and animal model were used to assess the function and mechanism of circFgfr2 in myogenesis and muscle regeneration in vitro or in vivo by RT‐qPCR, western blotting, dual‐luciferase activity assay, RNA immunoprecipitation, RNA fluorescence in situ hybridization, and chromatin immunoprecipitation. Results We profiled the dynamic circRNAome in pig skeletal muscle across 27 developmental stages and detected 52 918 high‐confidence circRNAs. A total of 2916 of these circRNAs are conserved across human, mouse, and pig, including four circRNAs (circFgfr2, circQrich1, circMettl9, and circCamta1) that were differentially expressed (|log2 fold change| > 1 and adjusted P value < 0.05) in various myogenesis systems. We further focused on a conserved circRNA produced from the fibroblast growth factor receptor 2 (Fgfr2) gene, termed circFgfr2, which was found to inhibit myoblast proliferation and promote differentiation and skeletal muscle regeneration. Mechanistically, circFgfr2 acted as a sponge for miR‐133 to regulate the mitogen‐activated protein kinase kinase kinase 20 (Map3k20) gene and JNK/MAPK pathway. Importantly, transcription factor Kruppel like factor 4 (Klf4), the downstream target of the JNK/MAPK pathway, directly bound to the promoter of circFgfr2 and affected its expression via an miR‐133/Map3k20/JNK/Klf4 auto‐regulatory feedback loop. RNA binding protein G3BP stress granule assembly factor 1 (G3bp1) inhibited the biogenesis of circFgfr2. Conclusions The present study provides a comprehensive circRNA resource for skeletal muscle study. The functional and mechanistic analysis of circFgfr2 uncovered a circRNA‐mediated auto‐regulatory feedback loop regulating myogenesis and muscle regeneration, which provides new insight to further understand the regulatory mechanism of circRNAs. |
| first_indexed | 2024-04-09T12:37:09Z |
| format | Article |
| id | doaj.art-0d1e52cf738d45d2ba3ae7799fa0389a |
| institution | Directory Open Access Journal |
| issn | 2190-5991 2190-6009 |
| language | English |
| last_indexed | 2024-04-09T12:37:09Z |
| publishDate | 2022-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Cachexia, Sarcopenia and Muscle |
| spelling | doaj.art-0d1e52cf738d45d2ba3ae7799fa0389a2023-05-15T09:27:16ZengWileyJournal of Cachexia, Sarcopenia and Muscle2190-59912190-60092022-02-0113169671210.1002/jcsm.12859circRNAome profiling reveals circFgfr2 regulates myogenesis and muscle regeneration via a feedback loopJunyu Yan0Yalan Yang1Xinhao Fan2Guoming Liang3Zishuai Wang4Jiju Li5Liyuan Wang6Yun Chen7Adeyinka Abiola Adetula8Yijie Tang9Kui Li10Dazhi Wang11Zhonglin Tang12Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen ChinaShenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen ChinaShenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen ChinaResearch Centre of Animal Nutritional Genomics, State Key Laboratory of Animal Nutrition, Institute of Animal Sciences Chinese Academy of Agricultural Sciences Shenzhen ChinaShenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen ChinaShenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen ChinaShenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen ChinaShenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen ChinaShenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen ChinaShenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen ChinaShenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen ChinaDepartment of Cardiology, Boston Children's Hospital Harvard Medical School Boston MA USAShenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen ChinaAbstract Background Circular RNAs (circRNAs) represent a novel class of non‐coding RNAs formed by a covalently closed loop and play crucial roles in many biological processes. Several circRNAs associated with myogenesis have been reported. However, the dynamic expression, function, and mechanism of circRNAs during myogenesis and skeletal muscle development are largely unknown. Methods Strand‐specific RNA‐sequencing (RNA‐seq) and microarray datasets were used to profile the dynamic circRNAome landscape during skeletal muscle development and myogenic differentiation. Bioinformatics analyses were used to characterize the circRNAome and identify candidate circRNAs associated with myogenesis. Bulk and single‐cell RNA‐seq were performed to identify the downstream genes and pathways of circFgfr2. The primary myoblast cells, C2C12 cells, and animal model were used to assess the function and mechanism of circFgfr2 in myogenesis and muscle regeneration in vitro or in vivo by RT‐qPCR, western blotting, dual‐luciferase activity assay, RNA immunoprecipitation, RNA fluorescence in situ hybridization, and chromatin immunoprecipitation. Results We profiled the dynamic circRNAome in pig skeletal muscle across 27 developmental stages and detected 52 918 high‐confidence circRNAs. A total of 2916 of these circRNAs are conserved across human, mouse, and pig, including four circRNAs (circFgfr2, circQrich1, circMettl9, and circCamta1) that were differentially expressed (|log2 fold change| > 1 and adjusted P value < 0.05) in various myogenesis systems. We further focused on a conserved circRNA produced from the fibroblast growth factor receptor 2 (Fgfr2) gene, termed circFgfr2, which was found to inhibit myoblast proliferation and promote differentiation and skeletal muscle regeneration. Mechanistically, circFgfr2 acted as a sponge for miR‐133 to regulate the mitogen‐activated protein kinase kinase kinase 20 (Map3k20) gene and JNK/MAPK pathway. Importantly, transcription factor Kruppel like factor 4 (Klf4), the downstream target of the JNK/MAPK pathway, directly bound to the promoter of circFgfr2 and affected its expression via an miR‐133/Map3k20/JNK/Klf4 auto‐regulatory feedback loop. RNA binding protein G3BP stress granule assembly factor 1 (G3bp1) inhibited the biogenesis of circFgfr2. Conclusions The present study provides a comprehensive circRNA resource for skeletal muscle study. The functional and mechanistic analysis of circFgfr2 uncovered a circRNA‐mediated auto‐regulatory feedback loop regulating myogenesis and muscle regeneration, which provides new insight to further understand the regulatory mechanism of circRNAs.https://doi.org/10.1002/jcsm.12859circRNAcircFgfr2Skeletal muscleDevelopmentRegenerationFeedback loop |
| spellingShingle | Junyu Yan Yalan Yang Xinhao Fan Guoming Liang Zishuai Wang Jiju Li Liyuan Wang Yun Chen Adeyinka Abiola Adetula Yijie Tang Kui Li Dazhi Wang Zhonglin Tang circRNAome profiling reveals circFgfr2 regulates myogenesis and muscle regeneration via a feedback loop Journal of Cachexia, Sarcopenia and Muscle circRNA circFgfr2 Skeletal muscle Development Regeneration Feedback loop |
| title | circRNAome profiling reveals circFgfr2 regulates myogenesis and muscle regeneration via a feedback loop |
| title_full | circRNAome profiling reveals circFgfr2 regulates myogenesis and muscle regeneration via a feedback loop |
| title_fullStr | circRNAome profiling reveals circFgfr2 regulates myogenesis and muscle regeneration via a feedback loop |
| title_full_unstemmed | circRNAome profiling reveals circFgfr2 regulates myogenesis and muscle regeneration via a feedback loop |
| title_short | circRNAome profiling reveals circFgfr2 regulates myogenesis and muscle regeneration via a feedback loop |
| title_sort | circrnaome profiling reveals circfgfr2 regulates myogenesis and muscle regeneration via a feedback loop |
| topic | circRNA circFgfr2 Skeletal muscle Development Regeneration Feedback loop |
| url | https://doi.org/10.1002/jcsm.12859 |
| work_keys_str_mv | AT junyuyan circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT yalanyang circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT xinhaofan circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT guomingliang circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT zishuaiwang circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT jijuli circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT liyuanwang circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT yunchen circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT adeyinkaabiolaadetula circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT yijietang circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT kuili circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT dazhiwang circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop AT zhonglintang circrnaomeprofilingrevealscircfgfr2regulatesmyogenesisandmuscleregenerationviaafeedbackloop |