Cas9/AAV9-Mediated Somatic Mutagenesis Uncovered the Cell-Autonomous Role of Sarcoplasmic/Endoplasmic Reticulum Calcium ATPase 2 in Murine Cardiomyocyte Maturation
Sarcoplasmic/endoplasmic reticulum calcium ATPase 2 (SERCA2) is a key player in cardiomyocyte calcium handling and also a classic target in the gene therapy for heart failure. SERCA2 expression dramatically increases during cardiomyocyte maturation in the postnatal phase of heart development, which...
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-04-01
|
| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2022.864516/full |
| _version_ | 1828872753879449600 |
|---|---|
| author | Junsen Lin Zhan Chen Luzi Yang Lei Liu Peng Yue Yueshen Sun Yueshen Sun Mingming Zhao Xiaoling Guo Xiaomin Hu Xiaomin Hu Yan Zhang Hong Zhang Yifei Li Yuxuan Guo Erdan Dong Erdan Dong |
| author_facet | Junsen Lin Zhan Chen Luzi Yang Lei Liu Peng Yue Yueshen Sun Yueshen Sun Mingming Zhao Xiaoling Guo Xiaomin Hu Xiaomin Hu Yan Zhang Hong Zhang Yifei Li Yuxuan Guo Erdan Dong Erdan Dong |
| author_sort | Junsen Lin |
| collection | DOAJ |
| description | Sarcoplasmic/endoplasmic reticulum calcium ATPase 2 (SERCA2) is a key player in cardiomyocyte calcium handling and also a classic target in the gene therapy for heart failure. SERCA2 expression dramatically increases during cardiomyocyte maturation in the postnatal phase of heart development, which is essential for the heart to acquire its full function in adults. However, whether and how SERCA2 regulates cardiomyocyte maturation remains unclear. Here, we performed Cas9/AAV9-mediated somatic mutagenesis (CASAAV) in mice and achieved cardiomyocyte-specific knockout of Atp2a2, the gene coding SERCA2. Through a cardiac genetic mosaic analysis, we demonstrated the cell-autonomous role of SERCA2 in building key ultrastructures of mature ventricular cardiomyocytes, including transverse-tubules and sarcomeres. SERCA2 also exerts a profound impact on oxidative respiration gene expression and sarcomere isoform switching from Myh7/Tnni1 to Myh6/Tnni3, which are transcriptional hallmarks of cardiomyocyte maturation. Together, this study uncovered a pivotal role of SERCA2 in heart development and provided new insights about SERCA2-based cardiac gene therapy. |
| first_indexed | 2024-12-13T07:00:55Z |
| format | Article |
| id | doaj.art-d32ccb3885124a17b7f843724dfec12f |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-12-13T07:00:55Z |
| publishDate | 2022-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-d32ccb3885124a17b7f843724dfec12f2022-12-21T23:55:56ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2022-04-011010.3389/fcell.2022.864516864516Cas9/AAV9-Mediated Somatic Mutagenesis Uncovered the Cell-Autonomous Role of Sarcoplasmic/Endoplasmic Reticulum Calcium ATPase 2 in Murine Cardiomyocyte MaturationJunsen Lin0Zhan Chen1Luzi Yang2Lei Liu3Peng Yue4Yueshen Sun5Yueshen Sun6Mingming Zhao7Xiaoling Guo8Xiaomin Hu9Xiaomin Hu10Yan Zhang11Hong Zhang12Yifei Li13Yuxuan Guo14Erdan Dong15Erdan Dong16Peking University Health Science Center, School of Basic Medical Sciences, The Institute of Cardiovascular Sciences, Key Laboratory of Molecular Cardiovascular Science of Ministry of Education, Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing, ChinaPeking University Health Science Center, School of Basic Medical Sciences, The Institute of Cardiovascular Sciences, Key Laboratory of Molecular Cardiovascular Science of Ministry of Education, Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing, ChinaPeking University Health Science Center, School of Basic Medical Sciences, The Institute of Cardiovascular Sciences, Key Laboratory of Molecular Cardiovascular Science of Ministry of Education, Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing, ChinaKey Laboratory of Birth Defects and Related Diseases of Women and Children of Ministry of Education (MOE), Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu, ChinaKey Laboratory of Birth Defects and Related Diseases of Women and Children of Ministry of Education (MOE), Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu, ChinaState Key Laboratory of Complex Severe and Rare Diseases, Department of Cardiology, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, ChinaDepartment of Medical Research Center, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, ChinaDepartment of Cardiology and Institute of Vascular Medicine, Peking University Third Hospital, National Health Commission of China (NHC) Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Beijing Key Laboratory of Cardiovascular Receptors Research. Beijing, ChinaBasic Medical Research Center, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, ChinaState Key Laboratory of Complex Severe and Rare Diseases, Department of Cardiology, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, ChinaDepartment of Medical Research Center, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, ChinaPeking University Health Science Center, School of Basic Medical Sciences, The Institute of Cardiovascular Sciences, Key Laboratory of Molecular Cardiovascular Science of Ministry of Education, Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing, ChinaPeking University Health Science Center, School of Basic Medical Sciences, The Institute of Cardiovascular Sciences, Key Laboratory of Molecular Cardiovascular Science of Ministry of Education, Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing, ChinaKey Laboratory of Birth Defects and Related Diseases of Women and Children of Ministry of Education (MOE), Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu, ChinaPeking University Health Science Center, School of Basic Medical Sciences, The Institute of Cardiovascular Sciences, Key Laboratory of Molecular Cardiovascular Science of Ministry of Education, Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing, ChinaPeking University Health Science Center, School of Basic Medical Sciences, The Institute of Cardiovascular Sciences, Key Laboratory of Molecular Cardiovascular Science of Ministry of Education, Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing, ChinaDepartment of Cardiology and Institute of Vascular Medicine, Peking University Third Hospital, National Health Commission of China (NHC) Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Beijing Key Laboratory of Cardiovascular Receptors Research. Beijing, ChinaSarcoplasmic/endoplasmic reticulum calcium ATPase 2 (SERCA2) is a key player in cardiomyocyte calcium handling and also a classic target in the gene therapy for heart failure. SERCA2 expression dramatically increases during cardiomyocyte maturation in the postnatal phase of heart development, which is essential for the heart to acquire its full function in adults. However, whether and how SERCA2 regulates cardiomyocyte maturation remains unclear. Here, we performed Cas9/AAV9-mediated somatic mutagenesis (CASAAV) in mice and achieved cardiomyocyte-specific knockout of Atp2a2, the gene coding SERCA2. Through a cardiac genetic mosaic analysis, we demonstrated the cell-autonomous role of SERCA2 in building key ultrastructures of mature ventricular cardiomyocytes, including transverse-tubules and sarcomeres. SERCA2 also exerts a profound impact on oxidative respiration gene expression and sarcomere isoform switching from Myh7/Tnni1 to Myh6/Tnni3, which are transcriptional hallmarks of cardiomyocyte maturation. Together, this study uncovered a pivotal role of SERCA2 in heart development and provided new insights about SERCA2-based cardiac gene therapy.https://www.frontiersin.org/articles/10.3389/fcell.2022.864516/fullcardiomyocyte maturationCas9/AAV9-mediated somatic mutagenesissarcoplasmic/endoplasmic reticulum calcium ATPase 2heart failuregenetic mosaic analysis |
| spellingShingle | Junsen Lin Zhan Chen Luzi Yang Lei Liu Peng Yue Yueshen Sun Yueshen Sun Mingming Zhao Xiaoling Guo Xiaomin Hu Xiaomin Hu Yan Zhang Hong Zhang Yifei Li Yuxuan Guo Erdan Dong Erdan Dong Cas9/AAV9-Mediated Somatic Mutagenesis Uncovered the Cell-Autonomous Role of Sarcoplasmic/Endoplasmic Reticulum Calcium ATPase 2 in Murine Cardiomyocyte Maturation Frontiers in Cell and Developmental Biology cardiomyocyte maturation Cas9/AAV9-mediated somatic mutagenesis sarcoplasmic/endoplasmic reticulum calcium ATPase 2 heart failure genetic mosaic analysis |
| title | Cas9/AAV9-Mediated Somatic Mutagenesis Uncovered the Cell-Autonomous Role of Sarcoplasmic/Endoplasmic Reticulum Calcium ATPase 2 in Murine Cardiomyocyte Maturation |
| title_full | Cas9/AAV9-Mediated Somatic Mutagenesis Uncovered the Cell-Autonomous Role of Sarcoplasmic/Endoplasmic Reticulum Calcium ATPase 2 in Murine Cardiomyocyte Maturation |
| title_fullStr | Cas9/AAV9-Mediated Somatic Mutagenesis Uncovered the Cell-Autonomous Role of Sarcoplasmic/Endoplasmic Reticulum Calcium ATPase 2 in Murine Cardiomyocyte Maturation |
| title_full_unstemmed | Cas9/AAV9-Mediated Somatic Mutagenesis Uncovered the Cell-Autonomous Role of Sarcoplasmic/Endoplasmic Reticulum Calcium ATPase 2 in Murine Cardiomyocyte Maturation |
| title_short | Cas9/AAV9-Mediated Somatic Mutagenesis Uncovered the Cell-Autonomous Role of Sarcoplasmic/Endoplasmic Reticulum Calcium ATPase 2 in Murine Cardiomyocyte Maturation |
| title_sort | cas9 aav9 mediated somatic mutagenesis uncovered the cell autonomous role of sarcoplasmic endoplasmic reticulum calcium atpase 2 in murine cardiomyocyte maturation |
| topic | cardiomyocyte maturation Cas9/AAV9-mediated somatic mutagenesis sarcoplasmic/endoplasmic reticulum calcium ATPase 2 heart failure genetic mosaic analysis |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2022.864516/full |
| work_keys_str_mv | AT junsenlin cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT zhanchen cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT luziyang cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT leiliu cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT pengyue cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT yueshensun cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT yueshensun cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT mingmingzhao cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT xiaolingguo cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT xiaominhu cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT xiaominhu cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT yanzhang cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT hongzhang cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT yifeili cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT yuxuanguo cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT erdandong cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration AT erdandong cas9aav9mediatedsomaticmutagenesisuncoveredthecellautonomousroleofsarcoplasmicendoplasmicreticulumcalciumatpase2inmurinecardiomyocytematuration |