Yuanxue Gao1,2,3, Shuang Wu1,2,3, Jiayue Pan1,2,3, Kai Zhang4, Xiaoyi Li1,2,3, Yangyang Xu5, Chentao Jin1,2,3, Xiao He1,2,3, Jingjing Shi1,2,3, Lijuan Ma1,2,3, Fujian Wu6, Yao Yao1,2,3, Ping Wang7,8, Qinggang He5, Feng Lan9, Hong Zhang10,11,12,13,14, Mei Tian15,16,17,18. 1. Department of Nuclear Medicine and PET-CT Center, The Second Hospital, Zhejiang University School of Medicine, Zhejiang, 310009, Hangzhou, China. 2. Key Laboratory of Medical Molecular Imaging of Zhejiang Province, Zhejiang, 310009, Hangzhou, China. 3. Institute of Nuclear Medicine and Molecular Imaging, Zhejiang University, Zhejiang, 310009, Hangzhou, China. 4. Laboratory for Pathophysiological and Health Science, RIKEN Center for Biosystems Dynamics Research, Kobe, Hyogo, 650-0047, Japan. 5. College of Chemical & Biological Engineering, Zhejiang University, Zhejiang, 310027, Hangzhou, China. 6. Beijing Laboratory for Cardiovascular Precision Medicine, The Key Laboratory of Remodeling-Related Cardiovascular Disease, Ministry of Education, Beijing Collaborative Innovation Center for Cardiovascular Disorders, Anzhen Hospital, Capital Medical University, Beijing, 100029, China. 7. Key Laboratory for Biomedical Engineering of Ministry of Education, Zhejiang University, Zhejiang, 310027, Hangzhou, China. 8. College of Biomedical Engineering and Instrument Science of Zhejiang University, Zhejiang, 310027, Hangzhou, China. 9. Beijing Laboratory for Cardiovascular Precision Medicine, The Key Laboratory of Remodeling-Related Cardiovascular Disease, Ministry of Education, Beijing Collaborative Innovation Center for Cardiovascular Disorders, Anzhen Hospital, Capital Medical University, Beijing, 100029, China. fenglan@ccmu.edu.cn. 10. Department of Nuclear Medicine and PET-CT Center, The Second Hospital, Zhejiang University School of Medicine, Zhejiang, 310009, Hangzhou, China. hzhang21@zju.edu.cn. 11. Key Laboratory of Medical Molecular Imaging of Zhejiang Province, Zhejiang, 310009, Hangzhou, China. hzhang21@zju.edu.cn. 12. Institute of Nuclear Medicine and Molecular Imaging, Zhejiang University, Zhejiang, 310009, Hangzhou, China. hzhang21@zju.edu.cn. 13. Key Laboratory for Biomedical Engineering of Ministry of Education, Zhejiang University, Zhejiang, 310027, Hangzhou, China. hzhang21@zju.edu.cn. 14. College of Biomedical Engineering and Instrument Science of Zhejiang University, Zhejiang, 310027, Hangzhou, China. hzhang21@zju.edu.cn. 15. Department of Nuclear Medicine and PET-CT Center, The Second Hospital, Zhejiang University School of Medicine, Zhejiang, 310009, Hangzhou, China. meitian@zju.edu.cn. 16. Key Laboratory of Medical Molecular Imaging of Zhejiang Province, Zhejiang, 310009, Hangzhou, China. meitian@zju.edu.cn. 17. Institute of Nuclear Medicine and Molecular Imaging, Zhejiang University, Zhejiang, 310009, Hangzhou, China. meitian@zju.edu.cn. 18. Key Laboratory for Biomedical Engineering of Ministry of Education, Zhejiang University, Zhejiang, 310027, Hangzhou, China. meitian@zju.edu.cn.
Abstract
PURPOSE: To investigate the post-transplantation behaviour and therapeutic efficacy of human urinary-induced pluripotent stem cell-derived cardiomyocytes (hUiCMs) in infarcted heart. METHODS: We used clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRISPR/Cas9) technology to integrate a triple-fusion (TF) reporter gene into the AAVS1 locus in human urine-derived hiPSCs (hUiPSCs) to generate TF-hUiPSCs that stably expressed monomeric red fluorescent protein for fluorescence imaging, firefly luciferase for bioluminescence imaging (BLI) and herpes simplex virus thymidine kinase for positron emission tomography (PET) imaging. RESULTS: Transplanted cardiomyocytes derived from TF-hUiPSCs (TF-hUiCMs) engrafted and proliferated in the infarcted heart as monitored by both BLI and PET imaging and significantly improved cardiac function. Under ischaemic conditions, TF-hUiCMs enhanced cardiomyocyte (CM) glucose metabolism and promoted angiogenic activity. CONCLUSION: This study established a CRISPR/Cas9-mediated multimodality reporter gene imaging system that can determine the dynamics and function of TF-hUiCMs in myocardial infarction, which is helpful for investigating the application of stem cell therapy.
PURPOSE: To investigate the post-transplantation behaviour and therapeutic efficacy of human urinary-induced pluripotent stem cell-derived cardiomyocytes (hUiCMs) in infarcted heart. METHODS: We used clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRISPR/Cas9) technology to integrate a triple-fusion (TF) reporter gene into the AAVS1 locus in human urine-derived hiPSCs (hUiPSCs) to generate TF-hUiPSCs that stably expressed monomeric red fluorescent protein for fluorescence imaging, firefly luciferase for bioluminescence imaging (BLI) and herpes simplex virus thymidine kinase for positron emission tomography (PET) imaging. RESULTS: Transplanted cardiomyocytes derived from TF-hUiPSCs (TF-hUiCMs) engrafted and proliferated in the infarcted heart as monitored by both BLI and PET imaging and significantly improved cardiac function. Under ischaemic conditions, TF-hUiCMs enhanced cardiomyocyte (CM) glucose metabolism and promoted angiogenic activity. CONCLUSION: This study established a CRISPR/Cas9-mediated multimodality reporter gene imaging system that can determine the dynamics and function of TF-hUiCMs in myocardial infarction, which is helpful for investigating the application of stem cell therapy.
Authors: Sang-Ging Ong; Bruno C Huber; Won Hee Lee; Kazuki Kodo; Antje D Ebert; Yu Ma; Patricia K Nguyen; Sebastian Diecke; Wen-Yi Chen; Joseph C Wu Journal: Circulation Date: 2015-08-25 Impact factor: 29.690
Authors: Paul Windisch; Daniel R Zwahlen; Frederik L Giesel; Eberhard Scholz; Patrick Lugenbiel; Jürgen Debus; Uwe Haberkorn; Sebastian Adeberg Journal: EJNMMI Res Date: 2021-02-19 Impact factor: 3.138