Engineered skeletal muscle recapitulates human muscle development, regeneration and dystrophy
Abstract Background Human pluripotent stem cell‐derived muscle models show great potential for translational research. Here, we describe developmentally inspired methods for the derivation of skeletal muscle cells and their utility in skeletal muscle tissue engineering with the aim to model skeletal...
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| Format: | Article |
| Language: | English |
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Wiley
2022-12-01
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| Series: | Journal of Cachexia, Sarcopenia and Muscle |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jcsm.13094 |
| _version_ | 1797204094932221952 |
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| author | Mina Shahriyari Md Rezaul Islam Sadman M. Sakib Malte Rinn Anastasia Rika Dennis Krüger Lalit Kaurani Verena Gisa Mandy Winterhoff Harithaa Anandakumar Orr Shomroni Matthias Schmidt Gabriela Salinas Andreas Unger Wolfgang A. Linke Jana Zschüntzsch Jens Schmidt Rhonda Bassel‐Duby Eric N. Olson André Fischer Wolfram‐Hubertus Zimmermann Malte Tiburcy |
| author_facet | Mina Shahriyari Md Rezaul Islam Sadman M. Sakib Malte Rinn Anastasia Rika Dennis Krüger Lalit Kaurani Verena Gisa Mandy Winterhoff Harithaa Anandakumar Orr Shomroni Matthias Schmidt Gabriela Salinas Andreas Unger Wolfgang A. Linke Jana Zschüntzsch Jens Schmidt Rhonda Bassel‐Duby Eric N. Olson André Fischer Wolfram‐Hubertus Zimmermann Malte Tiburcy |
| author_sort | Mina Shahriyari |
| collection | DOAJ |
| description | Abstract Background Human pluripotent stem cell‐derived muscle models show great potential for translational research. Here, we describe developmentally inspired methods for the derivation of skeletal muscle cells and their utility in skeletal muscle tissue engineering with the aim to model skeletal muscle regeneration and dystrophy in vitro. Methods Key steps include the directed differentiation of human pluripotent stem cells to embryonic muscle progenitors followed by primary and secondary foetal myogenesis into three‐dimensional muscle. To simulate Duchenne muscular dystrophy (DMD), a patient‐specific induced pluripotent stem cell line was compared to a CRISPR/Cas9‐edited isogenic control line. Results The established skeletal muscle differentiation protocol robustly and faithfully recapitulates critical steps of embryonic myogenesis in two‐dimensional and three‐dimensional cultures, resulting in functional human skeletal muscle organoids (SMOs) and engineered skeletal muscles (ESMs) with a regeneration‐competent satellite‐like cell pool. Tissue‐engineered muscle exhibits organotypic maturation and function (up to 5.7 ± 0.5 mN tetanic twitch tension at 100 Hz in ESM). Contractile performance could be further enhanced by timed thyroid hormone treatment, increasing the speed of contraction (time to peak contraction) as well as relaxation (time to 50% relaxation) of single twitches from 107 ± 2 to 75 ± 4 ms (P < 0.05) and from 146 ± 6 to 100 ± 6 ms (P < 0.05), respectively. Satellite‐like cells could be documented as largely quiescent PAX7+ cells (75 ± 6% Ki67−) located adjacent to muscle fibres confined under a laminin‐containing basal membrane. Activation of the engineered satellite‐like cell niche was documented in a cardiotoxin injury model with marked recovery of contractility to 57 ± 8% of the pre‐injury force 21 days post‐injury (P < 0.05 compared to Day 2 post‐injury), which was completely blocked by preceding irradiation. Absence of dystrophin in DMD ESM caused a marked reduction of contractile force (−35 ± 7%, P < 0.05) and impaired expression of fast myosin isoforms resulting in prolonged contraction (175 ± 14 ms, P < 0.05 vs. gene‐edited control) and relaxation (238 ± 22 ms, P < 0.05 vs. gene‐edited control) times. Restoration of dystrophin levels by gene editing rescued the DMD phenotype in ESM. Conclusions We introduce human muscle models with canonical properties of bona fide skeletal muscle in vivo to study muscle development, maturation, disease and repair. |
| first_indexed | 2024-04-11T13:58:54Z |
| format | Article |
| id | doaj.art-1634c6c809c344e3adcd3f61bb3b0fea |
| institution | Directory Open Access Journal |
| issn | 2190-5991 2190-6009 |
| language | English |
| last_indexed | 2024-04-24T08:29:46Z |
| publishDate | 2022-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Cachexia, Sarcopenia and Muscle |
| spelling | doaj.art-1634c6c809c344e3adcd3f61bb3b0fea2024-04-16T20:10:21ZengWileyJournal of Cachexia, Sarcopenia and Muscle2190-59912190-60092022-12-011363106312110.1002/jcsm.13094Engineered skeletal muscle recapitulates human muscle development, regeneration and dystrophyMina Shahriyari0Md Rezaul Islam1Sadman M. Sakib2Malte Rinn3Anastasia Rika4Dennis Krüger5Lalit Kaurani6Verena Gisa7Mandy Winterhoff8Harithaa Anandakumar9Orr Shomroni10Matthias Schmidt11Gabriela Salinas12Andreas Unger13Wolfgang A. Linke14Jana Zschüntzsch15Jens Schmidt16Rhonda Bassel‐Duby17Eric N. Olson18André Fischer19Wolfram‐Hubertus Zimmermann20Malte Tiburcy21Institute of Pharmacology and Toxicology University Medical Center Göttingen, Georg August University Göttingen GermanyDepartment for Epigenetics and Systems Medicine in Neurodegenerative Diseases German Center for Neurodegenerative Diseases (DZNE) Göttingen Göttingen GermanyDepartment for Epigenetics and Systems Medicine in Neurodegenerative Diseases German Center for Neurodegenerative Diseases (DZNE) Göttingen Göttingen GermanyInstitute of Pharmacology and Toxicology University Medical Center Göttingen, Georg August University Göttingen GermanyInstitute of Pharmacology and Toxicology University Medical Center Göttingen, Georg August University Göttingen GermanyDepartment for Epigenetics and Systems Medicine in Neurodegenerative Diseases German Center for Neurodegenerative Diseases (DZNE) Göttingen Göttingen GermanyDepartment for Epigenetics and Systems Medicine in Neurodegenerative Diseases German Center for Neurodegenerative Diseases (DZNE) Göttingen Göttingen GermanyDepartment for Epigenetics and Systems Medicine in Neurodegenerative Diseases German Center for Neurodegenerative Diseases (DZNE) Göttingen Göttingen GermanyInstitute of Pharmacology and Toxicology University Medical Center Göttingen, Georg August University Göttingen GermanyInstitute of Pharmacology and Toxicology University Medical Center Göttingen, Georg August University Göttingen GermanyNGS Integrative Genomics Core Unit, Institute of Human Genetics University Medical Center Göttingen, Georg August University Göttingen GermanyDepartment of Neurology, Neuromuscular Center University Medical Center Göttingen, Georg August University Göttingen GermanyNGS Integrative Genomics Core Unit, Institute of Human Genetics University Medical Center Göttingen, Georg August University Göttingen GermanyInstitute of Physiology II University of Münster Münster GermanyInstitute of Physiology II University of Münster Münster GermanyDepartment of Neurology, Neuromuscular Center University Medical Center Göttingen, Georg August University Göttingen GermanyDepartment of Neurology, Neuromuscular Center University Medical Center Göttingen, Georg August University Göttingen GermanyDepartment of Molecular Biology University of Texas Southwestern Medical Center Dallas TX USADepartment of Molecular Biology University of Texas Southwestern Medical Center Dallas TX USADepartment for Epigenetics and Systems Medicine in Neurodegenerative Diseases German Center for Neurodegenerative Diseases (DZNE) Göttingen Göttingen GermanyInstitute of Pharmacology and Toxicology University Medical Center Göttingen, Georg August University Göttingen GermanyInstitute of Pharmacology and Toxicology University Medical Center Göttingen, Georg August University Göttingen GermanyAbstract Background Human pluripotent stem cell‐derived muscle models show great potential for translational research. Here, we describe developmentally inspired methods for the derivation of skeletal muscle cells and their utility in skeletal muscle tissue engineering with the aim to model skeletal muscle regeneration and dystrophy in vitro. Methods Key steps include the directed differentiation of human pluripotent stem cells to embryonic muscle progenitors followed by primary and secondary foetal myogenesis into three‐dimensional muscle. To simulate Duchenne muscular dystrophy (DMD), a patient‐specific induced pluripotent stem cell line was compared to a CRISPR/Cas9‐edited isogenic control line. Results The established skeletal muscle differentiation protocol robustly and faithfully recapitulates critical steps of embryonic myogenesis in two‐dimensional and three‐dimensional cultures, resulting in functional human skeletal muscle organoids (SMOs) and engineered skeletal muscles (ESMs) with a regeneration‐competent satellite‐like cell pool. Tissue‐engineered muscle exhibits organotypic maturation and function (up to 5.7 ± 0.5 mN tetanic twitch tension at 100 Hz in ESM). Contractile performance could be further enhanced by timed thyroid hormone treatment, increasing the speed of contraction (time to peak contraction) as well as relaxation (time to 50% relaxation) of single twitches from 107 ± 2 to 75 ± 4 ms (P < 0.05) and from 146 ± 6 to 100 ± 6 ms (P < 0.05), respectively. Satellite‐like cells could be documented as largely quiescent PAX7+ cells (75 ± 6% Ki67−) located adjacent to muscle fibres confined under a laminin‐containing basal membrane. Activation of the engineered satellite‐like cell niche was documented in a cardiotoxin injury model with marked recovery of contractility to 57 ± 8% of the pre‐injury force 21 days post‐injury (P < 0.05 compared to Day 2 post‐injury), which was completely blocked by preceding irradiation. Absence of dystrophin in DMD ESM caused a marked reduction of contractile force (−35 ± 7%, P < 0.05) and impaired expression of fast myosin isoforms resulting in prolonged contraction (175 ± 14 ms, P < 0.05 vs. gene‐edited control) and relaxation (238 ± 22 ms, P < 0.05 vs. gene‐edited control) times. Restoration of dystrophin levels by gene editing rescued the DMD phenotype in ESM. Conclusions We introduce human muscle models with canonical properties of bona fide skeletal muscle in vivo to study muscle development, maturation, disease and repair.https://doi.org/10.1002/jcsm.13094Duchenne muscular dystrophyhypaxial dermomyotomelimb musclesatellite cellsskeletal muscle organoidsomite |
| spellingShingle | Mina Shahriyari Md Rezaul Islam Sadman M. Sakib Malte Rinn Anastasia Rika Dennis Krüger Lalit Kaurani Verena Gisa Mandy Winterhoff Harithaa Anandakumar Orr Shomroni Matthias Schmidt Gabriela Salinas Andreas Unger Wolfgang A. Linke Jana Zschüntzsch Jens Schmidt Rhonda Bassel‐Duby Eric N. Olson André Fischer Wolfram‐Hubertus Zimmermann Malte Tiburcy Engineered skeletal muscle recapitulates human muscle development, regeneration and dystrophy Journal of Cachexia, Sarcopenia and Muscle Duchenne muscular dystrophy hypaxial dermomyotome limb muscle satellite cells skeletal muscle organoid somite |
| title | Engineered skeletal muscle recapitulates human muscle development, regeneration and dystrophy |
| title_full | Engineered skeletal muscle recapitulates human muscle development, regeneration and dystrophy |
| title_fullStr | Engineered skeletal muscle recapitulates human muscle development, regeneration and dystrophy |
| title_full_unstemmed | Engineered skeletal muscle recapitulates human muscle development, regeneration and dystrophy |
| title_short | Engineered skeletal muscle recapitulates human muscle development, regeneration and dystrophy |
| title_sort | engineered skeletal muscle recapitulates human muscle development regeneration and dystrophy |
| topic | Duchenne muscular dystrophy hypaxial dermomyotome limb muscle satellite cells skeletal muscle organoid somite |
| url | https://doi.org/10.1002/jcsm.13094 |
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