The paradox of hnRNPK: both absence and excess impair skeletal muscle function in mice
Abstract Background The RNA-binding protein hnRNPK is essential for animal growth and development, with a particular emphasis in myogenesis. Despite its importance, the precise mechanisms by which hnRNPK influences skeletal muscle physiology and development remain inadequately characterized. Methods...
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2025-08-01
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| Series: | Skeletal Muscle |
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| Online Access: | https://doi.org/10.1186/s13395-025-00393-3 |
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| author | Yongjie Xu Yuxi Wang Xiaofang Cheng Mengjia Zhang Nuo Chen Jiahua Guo Yueru Huang Quanxi Li Tianyu Li Tiantian Meng Cencen Li Pengpeng Zhang Haixia Xu |
| author_facet | Yongjie Xu Yuxi Wang Xiaofang Cheng Mengjia Zhang Nuo Chen Jiahua Guo Yueru Huang Quanxi Li Tianyu Li Tiantian Meng Cencen Li Pengpeng Zhang Haixia Xu |
| author_sort | Yongjie Xu |
| collection | DOAJ |
| description | Abstract Background The RNA-binding protein hnRNPK is essential for animal growth and development, with a particular emphasis in myogenesis. Despite its importance, the precise mechanisms by which hnRNPK influences skeletal muscle physiology and development remain inadequately characterized. Methods To explore its regulatory function, we developed a Myf5-cre-mediated myoblast precursor-specific knockout mouse model (Hnrnpk mKO), an Acta1-CreEsr1-mediated myofiber-specific inducible knockout mouse model (Hnrnpk aKO), and an AAV9-mediated skeletal muscle-specific overexpression mouse model (AAV9-hnRNPK). Morphological alterations in skeletal muscle were assessed using hematoxylin and eosin (HE) staining subsequent to hnRNPK knockout or overexpression. Global gene expression changes in the tibialis anterior (TA) muscle were assessed via RNA sequencing (RNA-seq). Furthermore, reverse transcription quantitative polymerase chain reaction (RT-qPCR), western blot analysis, immunofluorescence, immunohistochemistry, co-immunoprecipitation (Co-IP), dual luciferase analysis, and reactive oxygen species (ROS) detection were utilized to elucidate the molecular mechanisms by which hnRNPK contributes to skeletal muscle development. Results Our findings indicate that the ablation of hnRNPK in myoblast precursors significantly impairs muscle development, disrupts fetal myogenesis, and results in embryonic lethality. In adult mice, both the loss and gain of hnRNPK function led to reduced muscle mass, decreased fiber size, and compromised skeletal muscle homeostasis. Importantly, the knockout of hnRNPK had a more substantial impact on skeletal muscle development compared to its overexpression, with myofiber-specific knockout leading to mortality within two weeks. Mechanistically, hnRNPK deficiency was associated with increased apoptosis and muscle atrophy, characterized by elevated expression of genes involved in apoptosis, muscle atrophy, and protein catabolism, along with impaired muscle contraction and extracellular matrix (ECM) organization. Conversely, hnRNPK overexpression was correlated with enhanced ferroptosis pathway and improved ECM organization, but was also associated with reduced oxidative phosphorylation and protein synthesis. The overexpression likely promotes ferroptosis via the hnRNPK/P53/Slc7a11/Gpx4 pathway, thereby accelerating muscle aging and reducing muscle mass. Conclusion In conclusion, our findings underscore the critical importance of precise hnRNPK expression levels in maintaining skeletal muscle health. Both deficiency and overexpression of hnRNPK disrupt skeletal muscle development, highlighting its pivotal role in muscle physiology. Clinical trial number Not applicable. |
| format | Article |
| id | doaj-art-679598719aa14a6f898efbdf18a4a982 |
| institution | Kabale University |
| issn | 2044-5040 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | BMC |
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| series | Skeletal Muscle |
| spelling | doaj-art-679598719aa14a6f898efbdf18a4a9822025-08-20T03:46:25ZengBMCSkeletal Muscle2044-50402025-08-0115111310.1186/s13395-025-00393-3The paradox of hnRNPK: both absence and excess impair skeletal muscle function in miceYongjie Xu0Yuxi Wang1Xiaofang Cheng2Mengjia Zhang3Nuo Chen4Jiahua Guo5Yueru Huang6Quanxi Li7Tianyu Li8Tiantian Meng9Cencen Li10Pengpeng Zhang11Haixia Xu12College of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityCollege of Life Science, Xinyang Normal UniversityAbstract Background The RNA-binding protein hnRNPK is essential for animal growth and development, with a particular emphasis in myogenesis. Despite its importance, the precise mechanisms by which hnRNPK influences skeletal muscle physiology and development remain inadequately characterized. Methods To explore its regulatory function, we developed a Myf5-cre-mediated myoblast precursor-specific knockout mouse model (Hnrnpk mKO), an Acta1-CreEsr1-mediated myofiber-specific inducible knockout mouse model (Hnrnpk aKO), and an AAV9-mediated skeletal muscle-specific overexpression mouse model (AAV9-hnRNPK). Morphological alterations in skeletal muscle were assessed using hematoxylin and eosin (HE) staining subsequent to hnRNPK knockout or overexpression. Global gene expression changes in the tibialis anterior (TA) muscle were assessed via RNA sequencing (RNA-seq). Furthermore, reverse transcription quantitative polymerase chain reaction (RT-qPCR), western blot analysis, immunofluorescence, immunohistochemistry, co-immunoprecipitation (Co-IP), dual luciferase analysis, and reactive oxygen species (ROS) detection were utilized to elucidate the molecular mechanisms by which hnRNPK contributes to skeletal muscle development. Results Our findings indicate that the ablation of hnRNPK in myoblast precursors significantly impairs muscle development, disrupts fetal myogenesis, and results in embryonic lethality. In adult mice, both the loss and gain of hnRNPK function led to reduced muscle mass, decreased fiber size, and compromised skeletal muscle homeostasis. Importantly, the knockout of hnRNPK had a more substantial impact on skeletal muscle development compared to its overexpression, with myofiber-specific knockout leading to mortality within two weeks. Mechanistically, hnRNPK deficiency was associated with increased apoptosis and muscle atrophy, characterized by elevated expression of genes involved in apoptosis, muscle atrophy, and protein catabolism, along with impaired muscle contraction and extracellular matrix (ECM) organization. Conversely, hnRNPK overexpression was correlated with enhanced ferroptosis pathway and improved ECM organization, but was also associated with reduced oxidative phosphorylation and protein synthesis. The overexpression likely promotes ferroptosis via the hnRNPK/P53/Slc7a11/Gpx4 pathway, thereby accelerating muscle aging and reducing muscle mass. Conclusion In conclusion, our findings underscore the critical importance of precise hnRNPK expression levels in maintaining skeletal muscle health. Both deficiency and overexpression of hnRNPK disrupt skeletal muscle development, highlighting its pivotal role in muscle physiology. Clinical trial number Not applicable.https://doi.org/10.1186/s13395-025-00393-3HnRNPKSkeletal muscleKnockoutOverexpressionMiceMuscle atrophy |
| spellingShingle | Yongjie Xu Yuxi Wang Xiaofang Cheng Mengjia Zhang Nuo Chen Jiahua Guo Yueru Huang Quanxi Li Tianyu Li Tiantian Meng Cencen Li Pengpeng Zhang Haixia Xu The paradox of hnRNPK: both absence and excess impair skeletal muscle function in mice Skeletal Muscle HnRNPK Skeletal muscle Knockout Overexpression Mice Muscle atrophy |
| title | The paradox of hnRNPK: both absence and excess impair skeletal muscle function in mice |
| title_full | The paradox of hnRNPK: both absence and excess impair skeletal muscle function in mice |
| title_fullStr | The paradox of hnRNPK: both absence and excess impair skeletal muscle function in mice |
| title_full_unstemmed | The paradox of hnRNPK: both absence and excess impair skeletal muscle function in mice |
| title_short | The paradox of hnRNPK: both absence and excess impair skeletal muscle function in mice |
| title_sort | paradox of hnrnpk both absence and excess impair skeletal muscle function in mice |
| topic | HnRNPK Skeletal muscle Knockout Overexpression Mice Muscle atrophy |
| url | https://doi.org/10.1186/s13395-025-00393-3 |
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