Disease modeling and gene correction of LGMDR21 iPSCs elucidates the role of POGLUT1 in skeletal muscle maintenance, regeneration, and the satellite cell niche
Autosomal recessive limb-girdle muscular dystrophy 21 (LGMDR21) is caused by pathogenic variants in protein O-glucosyltransferase 1 (POGLUT1), which is responsible for O-glucosylation of specific epidermal growth factor (EGF) repeats found in ∼50 mammalian proteins, including Notch receptors. Previo...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S216225312300210X |
| _version_ | 1797740998103662592 |
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| author | Jose L. Ortiz-Vitali Jianbo Wu Nasa Xu Annie W. Shieh Nima Niknejad Megumi Takeuchi Carmen Paradas Chunru Lin Hamed Jafar-Nejad Robert S. Haltiwanger Sidney H. Wang Radbod Darabi |
| author_facet | Jose L. Ortiz-Vitali Jianbo Wu Nasa Xu Annie W. Shieh Nima Niknejad Megumi Takeuchi Carmen Paradas Chunru Lin Hamed Jafar-Nejad Robert S. Haltiwanger Sidney H. Wang Radbod Darabi |
| author_sort | Jose L. Ortiz-Vitali |
| collection | DOAJ |
| description | Autosomal recessive limb-girdle muscular dystrophy 21 (LGMDR21) is caused by pathogenic variants in protein O-glucosyltransferase 1 (POGLUT1), which is responsible for O-glucosylation of specific epidermal growth factor (EGF) repeats found in ∼50 mammalian proteins, including Notch receptors. Previous data from patient biopsies indicated that impaired Notch signaling, reduction of muscle stem cells, and accelerated differentiation are probably involved in disease etiopathology. Using patient induced pluripotent stem cells (iPSCs), their corrected isotypes, and control iPSCs, gene expression profiling indicated dysregulation of POGLUT1, NOTCH, muscle development, extracellular matrix (ECM), cell adhesion, and migration as involved pathways. They also exhibited reduced in vitro POGLUT1 enzymatic activity and NOTCH signaling as well as defective myogenesis, proliferation, migration and differentiation. Furthermore, in vivo studies demonstrated significant reductions in engraftment, muscle stem cell formation, PAX7 expression, and maintenance, along with an increased percentage of mislocalized PAX7+ cells in the interstitial space. Gene correction in patient iPSCs using CRISPR-Cas9 nickase led to the rescue of the main in vitro and in vivo phenotypes. These results demonstrate the efficacy of iPSCs and gene correction in disease modeling and rescue of the phenotypes and provide evidence of the involvement of muscle stem cell niche localization, PAX7 expression, and cell migration as possible mechanisms in LGMDR21. |
| first_indexed | 2024-03-12T14:20:32Z |
| format | Article |
| id | doaj.art-890197d4a6154dcbb8fa3f4e093f2cfc |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-03-12T14:20:32Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-890197d4a6154dcbb8fa3f4e093f2cfc2023-08-19T04:31:59ZengElsevierMolecular Therapy: Nucleic Acids2162-25312023-09-0133683697Disease modeling and gene correction of LGMDR21 iPSCs elucidates the role of POGLUT1 in skeletal muscle maintenance, regeneration, and the satellite cell nicheJose L. Ortiz-Vitali0Jianbo Wu1Nasa Xu2Annie W. Shieh3Nima Niknejad4Megumi Takeuchi5Carmen Paradas6Chunru Lin7Hamed Jafar-Nejad8Robert S. Haltiwanger9Sidney H. Wang10Radbod Darabi11Center for Stem Cell and Regenerative Medicine (CSCRM), University of Texas Health Science Center at Houston, Houston, TX 77030, USACenter for Stem Cell and Regenerative Medicine (CSCRM), University of Texas Health Science Center at Houston, Houston, TX 77030, USACenter for Stem Cell and Regenerative Medicine (CSCRM), University of Texas Health Science Center at Houston, Houston, TX 77030, USACenter for Human Genetics, The Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases (IMM), University of Texas Health Science Center at Houston, Houston, TX 77030, USADepartment of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USAComplex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USANeurology Department, Neuromuscular Disorders Unit, Instituto de Biomedicina de Sevilla, Hospital U. Virgen Del Rocío, CSIC, Universidad de Sevilla, Avd. Manuel Siurot s/n, 41013 Sevilla, SpainDepartment of Molecular and Cellular Oncology, University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USAComplex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USACenter for Human Genetics, The Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases (IMM), University of Texas Health Science Center at Houston, Houston, TX 77030, USACenter for Stem Cell and Regenerative Medicine (CSCRM), University of Texas Health Science Center at Houston, Houston, TX 77030, USA; Corresponding author: Radbod Darabi, Center for Stem Cell and Regenerative Medicine (CSCRM), University of Texas Health Science Center at Houston, Houston, TX 77030, USA.Autosomal recessive limb-girdle muscular dystrophy 21 (LGMDR21) is caused by pathogenic variants in protein O-glucosyltransferase 1 (POGLUT1), which is responsible for O-glucosylation of specific epidermal growth factor (EGF) repeats found in ∼50 mammalian proteins, including Notch receptors. Previous data from patient biopsies indicated that impaired Notch signaling, reduction of muscle stem cells, and accelerated differentiation are probably involved in disease etiopathology. Using patient induced pluripotent stem cells (iPSCs), their corrected isotypes, and control iPSCs, gene expression profiling indicated dysregulation of POGLUT1, NOTCH, muscle development, extracellular matrix (ECM), cell adhesion, and migration as involved pathways. They also exhibited reduced in vitro POGLUT1 enzymatic activity and NOTCH signaling as well as defective myogenesis, proliferation, migration and differentiation. Furthermore, in vivo studies demonstrated significant reductions in engraftment, muscle stem cell formation, PAX7 expression, and maintenance, along with an increased percentage of mislocalized PAX7+ cells in the interstitial space. Gene correction in patient iPSCs using CRISPR-Cas9 nickase led to the rescue of the main in vitro and in vivo phenotypes. These results demonstrate the efficacy of iPSCs and gene correction in disease modeling and rescue of the phenotypes and provide evidence of the involvement of muscle stem cell niche localization, PAX7 expression, and cell migration as possible mechanisms in LGMDR21.http://www.sciencedirect.com/science/article/pii/S216225312300210XMT: RNA/DNA EditingPOGLUT1LGMDR21iPSCsgene correctionCRISPR-Cas9 |
| spellingShingle | Jose L. Ortiz-Vitali Jianbo Wu Nasa Xu Annie W. Shieh Nima Niknejad Megumi Takeuchi Carmen Paradas Chunru Lin Hamed Jafar-Nejad Robert S. Haltiwanger Sidney H. Wang Radbod Darabi Disease modeling and gene correction of LGMDR21 iPSCs elucidates the role of POGLUT1 in skeletal muscle maintenance, regeneration, and the satellite cell niche Molecular Therapy: Nucleic Acids MT: RNA/DNA Editing POGLUT1 LGMDR21 iPSCs gene correction CRISPR-Cas9 |
| title | Disease modeling and gene correction of LGMDR21 iPSCs elucidates the role of POGLUT1 in skeletal muscle maintenance, regeneration, and the satellite cell niche |
| title_full | Disease modeling and gene correction of LGMDR21 iPSCs elucidates the role of POGLUT1 in skeletal muscle maintenance, regeneration, and the satellite cell niche |
| title_fullStr | Disease modeling and gene correction of LGMDR21 iPSCs elucidates the role of POGLUT1 in skeletal muscle maintenance, regeneration, and the satellite cell niche |
| title_full_unstemmed | Disease modeling and gene correction of LGMDR21 iPSCs elucidates the role of POGLUT1 in skeletal muscle maintenance, regeneration, and the satellite cell niche |
| title_short | Disease modeling and gene correction of LGMDR21 iPSCs elucidates the role of POGLUT1 in skeletal muscle maintenance, regeneration, and the satellite cell niche |
| title_sort | disease modeling and gene correction of lgmdr21 ipscs elucidates the role of poglut1 in skeletal muscle maintenance regeneration and the satellite cell niche |
| topic | MT: RNA/DNA Editing POGLUT1 LGMDR21 iPSCs gene correction CRISPR-Cas9 |
| url | http://www.sciencedirect.com/science/article/pii/S216225312300210X |
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