CRISPR/Cas9-edited triple-fusion reporter gene imaging of dynamics and function of transplanted human urinary-induced pl
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ORIGINAL ARTICLE
CRISPR/Cas9-edited triple-fusion reporter gene imaging of dynamics and function of transplanted human urinary-induced pluripotent stem cell-derived cardiomyocytes Yuanxue Gao 1,2,3 & Shuang Wu 1,2,3 & Jiayue Pan 1,2,3 & Kai Zhang 4 & Xiaoyi Li 1,2,3 & Yangyang Xu 5 & Chentao Jin 1,2,3 & Xiao He 1,2,3 & Jingjing Shi 1,2,3 & Lijuan Ma 1,2,3 & Fujian Wu 6 & Yao Yao 1,2,3 & Ping Wang 7,8 & Qinggang He 5 & Feng Lan 6 & Hong Zhang 1,2,3,7,8 & Mei Tian 1,2,3,7 Received: 4 August 2020 / Accepted: 20 October 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
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, TFhUiCMs 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. Keywords Stem cells . CRISPR/Cas 9 . Molecular imaging . Myocardial infarction . Reporter gene
Yuanxue Gao, Shuang Wu, Jiayue Pan and Kai Zhang contributed equally to this work. This article is part of the Topical Collection on Preclinical Imaging * Mei Tian [email protected] * Hong Zhang [email protected] * Feng Lan [email protected]
4
Laboratory for Pathophysiological and Health Science, RIKEN Center for Biosystems Dynamics Research, Kobe, Hyogo 650-0047, Japan
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College of Chemical & Biological Engineering, Zhejiang University, Zhejiang 310027, Hangzhou, China
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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
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Department of Nuclear Medicine and PET-CT Center, The Second Hospital, Zhejiang University School of Medicine, Zhejiang 310009, Hangzhou, China
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Key Laboratory of Medical Molecular Imaging of Zhejiang Province, Zhejiang 310009, Hangzhou, China
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Key Laboratory for Biomedical Engineering of Ministry of Education, Zhejiang University, Zhejiang 3
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