SOX9 inactivation affects the proliferation and differentiation of human lung organoids
Abstract Background The regulation of the transcription factor sex-determining region Y-box transcription factor 9 (SOX9) in lung development has been described in mouse, but the same principles apply to human lung development is unknown due to a lack of appropriate experimental approaches and model...
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| Format: | Article |
| Language: | English |
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BMC
2021-06-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13287-021-02422-6 |
| _version_ | 1818644379547467776 |
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| author | Lian Li Jianqi Feng Shanshan Zhao Zhili Rong Ying Lin |
| author_facet | Lian Li Jianqi Feng Shanshan Zhao Zhili Rong Ying Lin |
| author_sort | Lian Li |
| collection | DOAJ |
| description | Abstract Background The regulation of the transcription factor sex-determining region Y-box transcription factor 9 (SOX9) in lung development has been described in mouse, but the same principles apply to human lung development is unknown due to a lack of appropriate experimental approaches and models. Methods Here, we used gene editing technology to inactivate SOX9 in human embryonic stem cells that were then induced to differentiate into lung organoids to investigate the role of SOX9 in human lung epithelium development. Results Complete knockout of the transactivation domain of SOX9 by gene editing resulted in indels in both alleles of SOX9. SOX9−/− hESCs could be induced to differentiate into lung progenitor organoids. In vitro long-term expansion showed that SOX9 inactivation did not affect the differentiation of pulmonary epithelial cells, but promoted apoptosis and reduced proliferative capacity in the organoids. When lung progenitor organoids were transplanted under the kidney capsule of immunodeficient mice, expression of the club cell marker secretoglobin family 1A member 1 (SCGB1A1) was detected in SOX9−/− transplants but was absent in wild-type (WT) transplants. The maturation of goblet cells was also affected by SOX9 inactivation, as evidenced by the presence of mucin 5 AC (MUC5AC) in the cytoplasm of SOX9−/− grafts as compared to WT grafts in which most MUC5AC was secreted into the lumen. In vivo lung orthotopic transplantations showed that SOX9 inactivation had a limited effect on the differentiation of alveolar cells and lung regeneration in injured mice. Conclusions SOX9 modulates the proliferative capacity of lung epithelium but is not an indispensable transcription factor in the regulation of human lung epithelium development. |
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| id | doaj.art-c4eb5be75a934186ae8eb33f9fff4736 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-17T00:13:55Z |
| publishDate | 2021-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-c4eb5be75a934186ae8eb33f9fff47362022-12-21T22:10:45ZengBMCStem Cell Research & Therapy1757-65122021-06-0112111210.1186/s13287-021-02422-6SOX9 inactivation affects the proliferation and differentiation of human lung organoidsLian Li0Jianqi Feng1Shanshan Zhao2Zhili Rong3Ying Lin4State Key Laboratory of Organ Failure Research, National Clinical Research Center of Kidney Disease, Key Laboratory of Organ Failure Research (Ministry of Education), Cancer Research Institute, School of Basic Medical Sciences, Southern Medical UniversityState Key Laboratory of Organ Failure Research, National Clinical Research Center of Kidney Disease, Key Laboratory of Organ Failure Research (Ministry of Education), Cancer Research Institute, School of Basic Medical Sciences, Southern Medical UniversityState Key Laboratory of Organ Failure Research, National Clinical Research Center of Kidney Disease, Key Laboratory of Organ Failure Research (Ministry of Education), Cancer Research Institute, School of Basic Medical Sciences, Southern Medical UniversityState Key Laboratory of Organ Failure Research, National Clinical Research Center of Kidney Disease, Key Laboratory of Organ Failure Research (Ministry of Education), Cancer Research Institute, School of Basic Medical Sciences, Southern Medical UniversityState Key Laboratory of Organ Failure Research, National Clinical Research Center of Kidney Disease, Key Laboratory of Organ Failure Research (Ministry of Education), Cancer Research Institute, School of Basic Medical Sciences, Southern Medical UniversityAbstract Background The regulation of the transcription factor sex-determining region Y-box transcription factor 9 (SOX9) in lung development has been described in mouse, but the same principles apply to human lung development is unknown due to a lack of appropriate experimental approaches and models. Methods Here, we used gene editing technology to inactivate SOX9 in human embryonic stem cells that were then induced to differentiate into lung organoids to investigate the role of SOX9 in human lung epithelium development. Results Complete knockout of the transactivation domain of SOX9 by gene editing resulted in indels in both alleles of SOX9. SOX9−/− hESCs could be induced to differentiate into lung progenitor organoids. In vitro long-term expansion showed that SOX9 inactivation did not affect the differentiation of pulmonary epithelial cells, but promoted apoptosis and reduced proliferative capacity in the organoids. When lung progenitor organoids were transplanted under the kidney capsule of immunodeficient mice, expression of the club cell marker secretoglobin family 1A member 1 (SCGB1A1) was detected in SOX9−/− transplants but was absent in wild-type (WT) transplants. The maturation of goblet cells was also affected by SOX9 inactivation, as evidenced by the presence of mucin 5 AC (MUC5AC) in the cytoplasm of SOX9−/− grafts as compared to WT grafts in which most MUC5AC was secreted into the lumen. In vivo lung orthotopic transplantations showed that SOX9 inactivation had a limited effect on the differentiation of alveolar cells and lung regeneration in injured mice. Conclusions SOX9 modulates the proliferative capacity of lung epithelium but is not an indispensable transcription factor in the regulation of human lung epithelium development.https://doi.org/10.1186/s13287-021-02422-6Lung organoidsSOX9CRISPR/Cas9ProliferationDifferentiation |
| spellingShingle | Lian Li Jianqi Feng Shanshan Zhao Zhili Rong Ying Lin SOX9 inactivation affects the proliferation and differentiation of human lung organoids Stem Cell Research & Therapy Lung organoids SOX9 CRISPR/Cas9 Proliferation Differentiation |
| title | SOX9 inactivation affects the proliferation and differentiation of human lung organoids |
| title_full | SOX9 inactivation affects the proliferation and differentiation of human lung organoids |
| title_fullStr | SOX9 inactivation affects the proliferation and differentiation of human lung organoids |
| title_full_unstemmed | SOX9 inactivation affects the proliferation and differentiation of human lung organoids |
| title_short | SOX9 inactivation affects the proliferation and differentiation of human lung organoids |
| title_sort | sox9 inactivation affects the proliferation and differentiation of human lung organoids |
| topic | Lung organoids SOX9 CRISPR/Cas9 Proliferation Differentiation |
| url | https://doi.org/10.1186/s13287-021-02422-6 |
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