CRISPR/Cas9-edited triple-fusion reporter gene imaging of dynamics and function of transplanted human urinary-induced pluripotent stem cell-derived cardiomyocytes

Yuanxue Gao; Shuang Wu; Jiayue Pan; Kai Zhang; Xiaoyi Li; Yangyang Xu; Chentao Jin; Xiao He; Jingjing Shi; Lijuan Ma; Fujian Wu; Yao Yao; Ping Wang; Qinggang He; Feng Lan; Hong Zhang; Mei Tian

doi:10.1007/s00259-020-05087-0

CRISPR/Cas9-edited triple-fusion reporter gene imaging of dynamics and function of transplanted human urinary-induced pluripotent stem cell-derived cardiomyocytes

Eur J Nucl Med Mol Imaging. 2021 Mar;48(3):708-720. doi: 10.1007/s00259-020-05087-0. Epub 2020 Nov 20.

Authors

Yuanxue Gao^#^{1

2

3}, Shuang Wu^#^{1

2

3}, Jiayue Pan^#^{1

2

3}, Kai Zhang^#⁴, Xiaoyi Li^{1

2

3}, Yangyang Xu⁵, Chentao Jin^{1

2

3}, Xiao He^{1

2

3}, Jingjing Shi^{1

2

3}, Lijuan Ma^{1

2

3}, Fujian Wu⁶, Yao Yao^{1

2

3}, Ping Wang^{7

8}, Qinggang He⁵, Feng Lan⁹, Hong Zhang^{10

11

12

13

14}, Mei Tian^{15

16

17

18}

Affiliations

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.

^# Contributed equally.

PMID: 33216174
DOI: 10.1007/s00259-020-05087-0

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.

Keywords: CRISPR/Cas 9; Molecular imaging; Myocardial infarction; Reporter gene; Stem cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems / genetics
Clustered Regularly Interspaced Short Palindromic Repeats
Genes, Reporter
Humans
Induced Pluripotent Stem Cells*
Myocytes, Cardiac

Grants and funding

81761148029, 81725009, 81900255/National Natural Science Foundation of China