Smad8 Is Increased in Duchenne Muscular Dystrophy and Suppresses miR-1, miR-133a, and miR-133b
Duchenne muscular dystrophy (DMD) is an X-linked recessive disease characterized by skeletal muscle instability, progressive muscle wasting, and fibrosis. A major driver of DMD pathology stems from aberrant upregulation of transforming growth factor β (TGFβ) signaling. In this report, we investigate...
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| Format: | Article |
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MDPI AG
2022-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/14/7515 |
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| author | Michael A. Lopez Ying Si Xianzhen Hu Valentyna Williams Fuad Qushair Jackson Carlyle Lyndsy Alesce Michael Conklin Shawn Gilbert Marcas M. Bamman Matthew S. Alexander Peter H. King |
| author_facet | Michael A. Lopez Ying Si Xianzhen Hu Valentyna Williams Fuad Qushair Jackson Carlyle Lyndsy Alesce Michael Conklin Shawn Gilbert Marcas M. Bamman Matthew S. Alexander Peter H. King |
| author_sort | Michael A. Lopez |
| collection | DOAJ |
| description | Duchenne muscular dystrophy (DMD) is an X-linked recessive disease characterized by skeletal muscle instability, progressive muscle wasting, and fibrosis. A major driver of DMD pathology stems from aberrant upregulation of transforming growth factor β (TGFβ) signaling. In this report, we investigated the major transducers of TGFβ signaling, i.e., receptor Smads (R-Smads), in DMD patient skeletal muscle and observed a 48-fold increase in <i>Smad8</i> mRNA. <i>Smad1</i>, <i>Smad2</i>, <i>Smad3</i>, and <i>Smad5</i> mRNA were only minimally increased. A similar pattern was observed in the muscle from the <i>mdx<sup>5cv</sup></i> mouse. Western blot analysis showed upregulation of phosphorylated Smad1, Smad5, and Smad8 compared to total Smad indicating activation of this pathway. In parallel, we observed a profound diminishment of muscle-enriched microRNAs (myomiRs): miR-1, miR-133a, and miR-133b. The pattern of Smad8 induction and myomiR suppression was recapitulated in C2C12 muscle cells after stimulation with bone morphogenetic protein 4 (BMP4), a signaling factor that we found upregulated in DMD muscle. Silencing <i>Smad8</i> in C2C12 myoblasts derepressed myomiRs and promoted myoblast differentiation; there was also a concomitant upregulation of myogenic regulatory factors (myogenin and myocyte enhancer factor 2D) and suppression of a pro-inflammatory cytokine (interleukin-6). Our data suggest that Smad8 is a negative regulator of miR-1, miR-133a, and miR-133b in muscle cells and that the BMP4-Smad8 axis is a driver of dystrophic pathology in DMD. |
| first_indexed | 2024-03-09T13:44:31Z |
| format | Article |
| id | doaj.art-b5a1840f1eb54c35931546ac52798328 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T13:44:31Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-b5a1840f1eb54c35931546ac527983282023-11-30T21:03:00ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-07-012314751510.3390/ijms23147515Smad8 Is Increased in Duchenne Muscular Dystrophy and Suppresses miR-1, miR-133a, and miR-133bMichael A. Lopez0Ying Si1Xianzhen Hu2Valentyna Williams3Fuad Qushair4Jackson Carlyle5Lyndsy Alesce6Michael Conklin7Shawn Gilbert8Marcas M. Bamman9Matthew S. Alexander10Peter H. King11Children’s of Alabama, Birmingham, AL 35233, USADepartment of Neurology, University of Alabama at Birmingham (UAB), Civitan 545C, 1530 3rd Avenue South, Birmingham, AL 35294, USADepartment of Pediatrics, University of Alabama at Birmingham (UAB), CHB314, 1600 7th Avenue South, Birmingham, AL 35233, USADepartment of Pediatrics, University of Alabama at Birmingham (UAB), CHB314, 1600 7th Avenue South, Birmingham, AL 35233, USADepartment of Pediatrics, University of Alabama at Birmingham (UAB), CHB314, 1600 7th Avenue South, Birmingham, AL 35233, USADepartment of Pediatrics, University of Alabama at Birmingham (UAB), CHB314, 1600 7th Avenue South, Birmingham, AL 35233, USADepartment of Neurology, University of Alabama at Birmingham (UAB), Civitan 545C, 1530 3rd Avenue South, Birmingham, AL 35294, USAChildren’s of Alabama, Birmingham, AL 35233, USAChildren’s of Alabama, Birmingham, AL 35233, USADepartment of Neurology, University of Alabama at Birmingham (UAB), Civitan 545C, 1530 3rd Avenue South, Birmingham, AL 35294, USAChildren’s of Alabama, Birmingham, AL 35233, USADepartment of Neurology, University of Alabama at Birmingham (UAB), Civitan 545C, 1530 3rd Avenue South, Birmingham, AL 35294, USADuchenne muscular dystrophy (DMD) is an X-linked recessive disease characterized by skeletal muscle instability, progressive muscle wasting, and fibrosis. A major driver of DMD pathology stems from aberrant upregulation of transforming growth factor β (TGFβ) signaling. In this report, we investigated the major transducers of TGFβ signaling, i.e., receptor Smads (R-Smads), in DMD patient skeletal muscle and observed a 48-fold increase in <i>Smad8</i> mRNA. <i>Smad1</i>, <i>Smad2</i>, <i>Smad3</i>, and <i>Smad5</i> mRNA were only minimally increased. A similar pattern was observed in the muscle from the <i>mdx<sup>5cv</sup></i> mouse. Western blot analysis showed upregulation of phosphorylated Smad1, Smad5, and Smad8 compared to total Smad indicating activation of this pathway. In parallel, we observed a profound diminishment of muscle-enriched microRNAs (myomiRs): miR-1, miR-133a, and miR-133b. The pattern of Smad8 induction and myomiR suppression was recapitulated in C2C12 muscle cells after stimulation with bone morphogenetic protein 4 (BMP4), a signaling factor that we found upregulated in DMD muscle. Silencing <i>Smad8</i> in C2C12 myoblasts derepressed myomiRs and promoted myoblast differentiation; there was also a concomitant upregulation of myogenic regulatory factors (myogenin and myocyte enhancer factor 2D) and suppression of a pro-inflammatory cytokine (interleukin-6). Our data suggest that Smad8 is a negative regulator of miR-1, miR-133a, and miR-133b in muscle cells and that the BMP4-Smad8 axis is a driver of dystrophic pathology in DMD.https://www.mdpi.com/1422-0067/23/14/7515BMP4DuchennemiRNAmuscleSmad8 |
| spellingShingle | Michael A. Lopez Ying Si Xianzhen Hu Valentyna Williams Fuad Qushair Jackson Carlyle Lyndsy Alesce Michael Conklin Shawn Gilbert Marcas M. Bamman Matthew S. Alexander Peter H. King Smad8 Is Increased in Duchenne Muscular Dystrophy and Suppresses miR-1, miR-133a, and miR-133b International Journal of Molecular Sciences BMP4 Duchenne miRNA muscle Smad8 |
| title | Smad8 Is Increased in Duchenne Muscular Dystrophy and Suppresses miR-1, miR-133a, and miR-133b |
| title_full | Smad8 Is Increased in Duchenne Muscular Dystrophy and Suppresses miR-1, miR-133a, and miR-133b |
| title_fullStr | Smad8 Is Increased in Duchenne Muscular Dystrophy and Suppresses miR-1, miR-133a, and miR-133b |
| title_full_unstemmed | Smad8 Is Increased in Duchenne Muscular Dystrophy and Suppresses miR-1, miR-133a, and miR-133b |
| title_short | Smad8 Is Increased in Duchenne Muscular Dystrophy and Suppresses miR-1, miR-133a, and miR-133b |
| title_sort | smad8 is increased in duchenne muscular dystrophy and suppresses mir 1 mir 133a and mir 133b |
| topic | BMP4 Duchenne miRNA muscle Smad8 |
| url | https://www.mdpi.com/1422-0067/23/14/7515 |
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