Hypoxia enhances human myoblast differentiation: involvement of HIF1α and impact of DUX4, the FSHD causal gene
Abstract Background Hypoxia is known to modify skeletal muscle biological functions and muscle regeneration. However, the mechanisms underlying the effects of hypoxia on human myoblast differentiation remain unclear. The hypoxic response pathway is of particular interest in patients with hereditary...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-12-01
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| Series: | Skeletal Muscle |
| Online Access: | https://doi.org/10.1186/s13395-023-00330-2 |
| _version_ | 1827581289195110400 |
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| author | Thuy-Hang Nguyen Lise Paprzycki Alexandre Legrand Anne-Emilie Declèves Philipp Heher Maelle Limpens Alexandra Belayew Christopher R. S. Banerji Peter S. Zammit Alexandra Tassin |
| author_facet | Thuy-Hang Nguyen Lise Paprzycki Alexandre Legrand Anne-Emilie Declèves Philipp Heher Maelle Limpens Alexandra Belayew Christopher R. S. Banerji Peter S. Zammit Alexandra Tassin |
| author_sort | Thuy-Hang Nguyen |
| collection | DOAJ |
| description | Abstract Background Hypoxia is known to modify skeletal muscle biological functions and muscle regeneration. However, the mechanisms underlying the effects of hypoxia on human myoblast differentiation remain unclear. The hypoxic response pathway is of particular interest in patients with hereditary muscular dystrophies since many present respiratory impairment and muscle regeneration defects. For example, an altered hypoxia response characterizes the muscles of patients with facioscapulohumeral dystrophy (FSHD). Methods We examined the impact of hypoxia on the differentiation of human immortalized myoblasts (LHCN-M2) cultured in normoxia (PO2: 21%) or hypoxia (PO2: 1%). Cells were grown in proliferation (myoblasts) or differentiation medium for 2 (myocytes) or 4 days (myotubes). We evaluated proliferation rate by EdU incorporation, used myogenin-positive nuclei as a differentiation marker for myocytes, and determined the fusion index and myosin heavy chain-positive area in myotubes. The contribution of HIF1α was studied by gain (CoCl2) and loss (siRNAs) of function experiments. We further examined hypoxia in LHCN-M2-iDUX4 myoblasts with inducible expression of DUX4, the transcription factor underlying FSHD pathology. Results We found that the hypoxic response did not impact myoblast proliferation but activated precocious myogenic differentiation and that HIF1α was critical for this process. Hypoxia also enhanced the late differentiation of human myocytes, but in an HIF1α-independent manner. Interestingly, the impact of hypoxia on muscle cell proliferation was influenced by dexamethasone. In the FSHD pathological context, DUX4 suppressed HIF1α-mediated precocious muscle differentiation. Conclusion Hypoxia stimulates myogenic differentiation in healthy myoblasts, with HIF1α-dependent early steps. In FSHD, DUX4-HIF1α interplay indicates a novel mechanism by which DUX4 could interfere with HIF1α function in the myogenic program and therefore with FSHD muscle performance and regeneration. |
| first_indexed | 2024-03-08T22:34:15Z |
| format | Article |
| id | doaj.art-54f7e8b9df5a4c7f8986eef5f8194eb0 |
| institution | Directory Open Access Journal |
| issn | 2044-5040 |
| language | English |
| last_indexed | 2024-03-08T22:34:15Z |
| publishDate | 2023-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Skeletal Muscle |
| spelling | doaj.art-54f7e8b9df5a4c7f8986eef5f8194eb02023-12-17T12:33:57ZengBMCSkeletal Muscle2044-50402023-12-0113111410.1186/s13395-023-00330-2Hypoxia enhances human myoblast differentiation: involvement of HIF1α and impact of DUX4, the FSHD causal geneThuy-Hang Nguyen0Lise Paprzycki1Alexandre Legrand2Anne-Emilie Declèves3Philipp Heher4Maelle Limpens5Alexandra Belayew6Christopher R. S. Banerji7Peter S. Zammit8Alexandra Tassin9Laboratory of Respiratory Physiology, Pathophysiology and Rehabilitation, Research Institute for Health Sciences and Technology, University of MonsLaboratory of Respiratory Physiology, Pathophysiology and Rehabilitation, Research Institute for Health Sciences and Technology, University of MonsLaboratory of Respiratory Physiology, Pathophysiology and Rehabilitation, Research Institute for Health Sciences and Technology, University of MonsDepartment of Metabolic and Molecular Biochemistry, Research Institute for Health Sciences and Technology, University of MonsRandall Centre for Cell and Molecular Biophysics, King’s College London, Guy’s CampusLaboratory of Respiratory Physiology, Pathophysiology and Rehabilitation, Research Institute for Health Sciences and Technology, University of MonsLaboratory of Respiratory Physiology, Pathophysiology and Rehabilitation, Research Institute for Health Sciences and Technology, University of MonsRandall Centre for Cell and Molecular Biophysics, King’s College London, Guy’s CampusRandall Centre for Cell and Molecular Biophysics, King’s College London, Guy’s CampusLaboratory of Respiratory Physiology, Pathophysiology and Rehabilitation, Research Institute for Health Sciences and Technology, University of MonsAbstract Background Hypoxia is known to modify skeletal muscle biological functions and muscle regeneration. However, the mechanisms underlying the effects of hypoxia on human myoblast differentiation remain unclear. The hypoxic response pathway is of particular interest in patients with hereditary muscular dystrophies since many present respiratory impairment and muscle regeneration defects. For example, an altered hypoxia response characterizes the muscles of patients with facioscapulohumeral dystrophy (FSHD). Methods We examined the impact of hypoxia on the differentiation of human immortalized myoblasts (LHCN-M2) cultured in normoxia (PO2: 21%) or hypoxia (PO2: 1%). Cells were grown in proliferation (myoblasts) or differentiation medium for 2 (myocytes) or 4 days (myotubes). We evaluated proliferation rate by EdU incorporation, used myogenin-positive nuclei as a differentiation marker for myocytes, and determined the fusion index and myosin heavy chain-positive area in myotubes. The contribution of HIF1α was studied by gain (CoCl2) and loss (siRNAs) of function experiments. We further examined hypoxia in LHCN-M2-iDUX4 myoblasts with inducible expression of DUX4, the transcription factor underlying FSHD pathology. Results We found that the hypoxic response did not impact myoblast proliferation but activated precocious myogenic differentiation and that HIF1α was critical for this process. Hypoxia also enhanced the late differentiation of human myocytes, but in an HIF1α-independent manner. Interestingly, the impact of hypoxia on muscle cell proliferation was influenced by dexamethasone. In the FSHD pathological context, DUX4 suppressed HIF1α-mediated precocious muscle differentiation. Conclusion Hypoxia stimulates myogenic differentiation in healthy myoblasts, with HIF1α-dependent early steps. In FSHD, DUX4-HIF1α interplay indicates a novel mechanism by which DUX4 could interfere with HIF1α function in the myogenic program and therefore with FSHD muscle performance and regeneration.https://doi.org/10.1186/s13395-023-00330-2 |
| spellingShingle | Thuy-Hang Nguyen Lise Paprzycki Alexandre Legrand Anne-Emilie Declèves Philipp Heher Maelle Limpens Alexandra Belayew Christopher R. S. Banerji Peter S. Zammit Alexandra Tassin Hypoxia enhances human myoblast differentiation: involvement of HIF1α and impact of DUX4, the FSHD causal gene Skeletal Muscle |
| title | Hypoxia enhances human myoblast differentiation: involvement of HIF1α and impact of DUX4, the FSHD causal gene |
| title_full | Hypoxia enhances human myoblast differentiation: involvement of HIF1α and impact of DUX4, the FSHD causal gene |
| title_fullStr | Hypoxia enhances human myoblast differentiation: involvement of HIF1α and impact of DUX4, the FSHD causal gene |
| title_full_unstemmed | Hypoxia enhances human myoblast differentiation: involvement of HIF1α and impact of DUX4, the FSHD causal gene |
| title_short | Hypoxia enhances human myoblast differentiation: involvement of HIF1α and impact of DUX4, the FSHD causal gene |
| title_sort | hypoxia enhances human myoblast differentiation involvement of hif1α and impact of dux4 the fshd causal gene |
| url | https://doi.org/10.1186/s13395-023-00330-2 |
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