Inducible caspase-9 suicide gene under control of endogenous oct4 to safeguard mouse and human pluripotent stem cell therapy
Pluripotent stem cells (PSCs) are promising in regenerative medicine. A major challenge of PSC therapy is the risk of teratoma formation because of the contamination of undifferentiated stem cells. Constitutive promoters or endogenous SOX2 promoters have been used to drive inducible caspase-9 (iCasp...
| Main Authors: | , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-03-01
|
| Series: | Molecular Therapy: Methods & Clinical Development |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050122000158 |
| _version_ | 1819349528658051072 |
|---|---|
| author | Yang Liu Yang Yang Yangyang Suo Chuan Li Min Chen Shuwen Zheng Hao Li Chengcheng Tang Nana Fan Ting Lan Jizeng Zhou Yingying Li Jiaowei Wang Huangyao Chen Qingjian Zou Liangxue Lai |
| author_facet | Yang Liu Yang Yang Yangyang Suo Chuan Li Min Chen Shuwen Zheng Hao Li Chengcheng Tang Nana Fan Ting Lan Jizeng Zhou Yingying Li Jiaowei Wang Huangyao Chen Qingjian Zou Liangxue Lai |
| author_sort | Yang Liu |
| collection | DOAJ |
| description | Pluripotent stem cells (PSCs) are promising in regenerative medicine. A major challenge of PSC therapy is the risk of teratoma formation because of the contamination of undifferentiated stem cells. Constitutive promoters or endogenous SOX2 promoters have been used to drive inducible caspase-9 (iCasp9) gene expression but cannot specifically eradicate undifferentiated PSCs. Here, we inserted iCasp9 gene into the endogenous OCT4 locus of human and mouse PSCs without affecting their pluripotency. A chemical inducer of dimerization (CID), AP1903, induced iCasp9 activation, which led to the apoptosis of specific undifferentiated PSCs in vitro and in vivo. Differentiated cell lineages survived because of the silence of the endogenous OCT4 gene. Human and mouse PSCs were controllable when CID was administrated within 2 weeks after PSC injection in immunodeficient mice. However, an interval longer than 2 weeks caused teratoma formation and mouse death because a mass of somatic cells already differentiated from the PSCs. In conclusion, we have developed a specific and efficient PSC suicide system that will be of value in the clinical applications of PSC-based therapy. |
| first_indexed | 2024-12-24T19:01:58Z |
| format | Article |
| id | doaj.art-816ae9b9e7cd405baf12a610222efcf9 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-24T19:01:58Z |
| publishDate | 2022-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-816ae9b9e7cd405baf12a610222efcf92022-12-21T16:43:11ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012022-03-0124332341Inducible caspase-9 suicide gene under control of endogenous oct4 to safeguard mouse and human pluripotent stem cell therapyYang Liu0Yang Yang1Yangyang Suo2Chuan Li3Min Chen4Shuwen Zheng5Hao Li6Chengcheng Tang7Nana Fan8Ting Lan9Jizeng Zhou10Yingying Li11Jiaowei Wang12Huangyao Chen13Qingjian Zou14Liangxue Lai15School of Life Sciences, University of Science and Technology of China, Hefei 230026, China; Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Biotechnology and Health Sciences, Wuyi University, Jiangmen 529020, China; CAS Key Laboratory of Regenerative Biology, Guangdong-Hong Kong Stem Cell and Regenerative Medicine Research Centre, Joint Research Laboratory on Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, ChinaSchool of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, ChinaJoint School of Life Science, Guangzhou Institutes of Biomedicine and Health, Chinese Academic and Sciences, Guangzhou Medical University, Guangzhou 511495, ChinaGuangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Biotechnology and Health Sciences, Wuyi University, Jiangmen 529020, ChinaGuangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Biotechnology and Health Sciences, Wuyi University, Jiangmen 529020, ChinaGuangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Biotechnology and Health Sciences, Wuyi University, Jiangmen 529020, ChinaCAS Key Laboratory of Regenerative Biology, Guangdong-Hong Kong Stem Cell and Regenerative Medicine Research Centre, Joint Research Laboratory on Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, ChinaGuangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Biotechnology and Health Sciences, Wuyi University, Jiangmen 529020, ChinaCAS Key Laboratory of Regenerative Biology, Guangdong-Hong Kong Stem Cell and Regenerative Medicine Research Centre, Joint Research Laboratory on Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, ChinaCAS Key Laboratory of Regenerative Biology, Guangdong-Hong Kong Stem Cell and Regenerative Medicine Research Centre, Joint Research Laboratory on Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, ChinaSchool of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, ChinaCAS Key Laboratory of Regenerative Biology, Guangdong-Hong Kong Stem Cell and Regenerative Medicine Research Centre, Joint Research Laboratory on Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, ChinaCAS Key Laboratory of Regenerative Biology, Guangdong-Hong Kong Stem Cell and Regenerative Medicine Research Centre, Joint Research Laboratory on Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, ChinaCAS Key Laboratory of Regenerative Biology, Guangdong-Hong Kong Stem Cell and Regenerative Medicine Research Centre, Joint Research Laboratory on Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, ChinaGuangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Biotechnology and Health Sciences, Wuyi University, Jiangmen 529020, China; Corresponding author: Qingjian Zou, Guangdong Provincial Key Laboratory of Large Animal models for Biomedicine, School of Biotechnology and Health Sciences, Wuyi University, Jiangmen 529020, China.School of Life Sciences, University of Science and Technology of China, Hefei 230026, China; Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Biotechnology and Health Sciences, Wuyi University, Jiangmen 529020, China; CAS Key Laboratory of Regenerative Biology, Guangdong-Hong Kong Stem Cell and Regenerative Medicine Research Centre, Joint Research Laboratory on Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; Bioland Laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangzhou 510005, China; Research Unit of Generation of Large Animal Disease Models, Chinese Academy of Medical Sciences (2019RU015), Guangzhou 510530, China; Corresponding author: Liangxue Lai, School of Life Sciences, University of Science and Technology of China, Hefei 230026, China.Pluripotent stem cells (PSCs) are promising in regenerative medicine. A major challenge of PSC therapy is the risk of teratoma formation because of the contamination of undifferentiated stem cells. Constitutive promoters or endogenous SOX2 promoters have been used to drive inducible caspase-9 (iCasp9) gene expression but cannot specifically eradicate undifferentiated PSCs. Here, we inserted iCasp9 gene into the endogenous OCT4 locus of human and mouse PSCs without affecting their pluripotency. A chemical inducer of dimerization (CID), AP1903, induced iCasp9 activation, which led to the apoptosis of specific undifferentiated PSCs in vitro and in vivo. Differentiated cell lineages survived because of the silence of the endogenous OCT4 gene. Human and mouse PSCs were controllable when CID was administrated within 2 weeks after PSC injection in immunodeficient mice. However, an interval longer than 2 weeks caused teratoma formation and mouse death because a mass of somatic cells already differentiated from the PSCs. In conclusion, we have developed a specific and efficient PSC suicide system that will be of value in the clinical applications of PSC-based therapy.http://www.sciencedirect.com/science/article/pii/S2329050122000158pluripotent stem cells (PSCs)inducible caspase-9 (iCasp9) geneapoptosisOCT4 geneteratoma formation |
| spellingShingle | Yang Liu Yang Yang Yangyang Suo Chuan Li Min Chen Shuwen Zheng Hao Li Chengcheng Tang Nana Fan Ting Lan Jizeng Zhou Yingying Li Jiaowei Wang Huangyao Chen Qingjian Zou Liangxue Lai Inducible caspase-9 suicide gene under control of endogenous oct4 to safeguard mouse and human pluripotent stem cell therapy Molecular Therapy: Methods & Clinical Development pluripotent stem cells (PSCs) inducible caspase-9 (iCasp9) gene apoptosis OCT4 gene teratoma formation |
| title | Inducible caspase-9 suicide gene under control of endogenous oct4 to safeguard mouse and human pluripotent stem cell therapy |
| title_full | Inducible caspase-9 suicide gene under control of endogenous oct4 to safeguard mouse and human pluripotent stem cell therapy |
| title_fullStr | Inducible caspase-9 suicide gene under control of endogenous oct4 to safeguard mouse and human pluripotent stem cell therapy |
| title_full_unstemmed | Inducible caspase-9 suicide gene under control of endogenous oct4 to safeguard mouse and human pluripotent stem cell therapy |
| title_short | Inducible caspase-9 suicide gene under control of endogenous oct4 to safeguard mouse and human pluripotent stem cell therapy |
| title_sort | inducible caspase 9 suicide gene under control of endogenous oct4 to safeguard mouse and human pluripotent stem cell therapy |
| topic | pluripotent stem cells (PSCs) inducible caspase-9 (iCasp9) gene apoptosis OCT4 gene teratoma formation |
| url | http://www.sciencedirect.com/science/article/pii/S2329050122000158 |
| work_keys_str_mv | AT yangliu induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT yangyang induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT yangyangsuo induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT chuanli induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT minchen induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT shuwenzheng induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT haoli induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT chengchengtang induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT nanafan induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT tinglan induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT jizengzhou induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT yingyingli induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT jiaoweiwang induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT huangyaochen induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT qingjianzou induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy AT liangxuelai induciblecaspase9suicidegeneundercontrolofendogenousoct4tosafeguardmouseandhumanpluripotentstemcelltherapy |