CRISPR/Cas9-mediated introduction of the sodium/iodide symporter gene enables noninvasive in vivo tracking of induced pluripotent stem cell-derived cardiomyocytes

John W Ostrominski; Ravi Chandra Yada; Noriko Sato; Michael Klein; Ksenia Blinova; Dakshesh Patel; Racquel Valadez; Maryknoll Palisoc; Stefania Pittaluga; Kah-Whye Peng; Hong San; Yongshun Lin; Falguni Basuli; Xiang Zhang; Rolf E Swenson; Mark Haigney; Peter L Choyke; Jizhong Zou; Manfred Boehm; So Gun Hong; Cynthia E Dunbar

doi:10.1002/sctm.20-0019

CRISPR/Cas9-mediated introduction of the sodium/iodide symporter gene enables noninvasive in vivo tracking of induced pluripotent stem cell-derived cardiomyocytes

Stem Cells Transl Med. 2020 Oct;9(10):1203-1217. doi: 10.1002/sctm.20-0019. Epub 2020 Jul 23.

Authors

John W Ostrominski¹, Ravi Chandra Yada¹, Noriko Sato², Michael Klein³, Ksenia Blinova⁴, Dakshesh Patel⁴, Racquel Valadez⁵, Maryknoll Palisoc⁵, Stefania Pittaluga⁵, Kah-Whye Peng⁶, Hong San⁷, Yongshun Lin⁸, Falguni Basuli⁹, Xiang Zhang⁹, Rolf E Swenson⁹, Mark Haigney³, Peter L Choyke², Jizhong Zou⁸, Manfred Boehm¹⁰, So Gun Hong¹, Cynthia E Dunbar¹

Affiliations

¹ Translational Stem Cell Biology Branch, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health (NIH), Bethesda, Maryland, USA.
² Molecular Imaging Program, Laboratory of Cellular Therapeutics, National Cancer Institute (NCI), NIH, Bethesda, Maryland, USA.
³ Division of Cardiology, Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.
⁴ Office of Science and Engineering Laboratories, Center for Devices and Radiological Health, US Food and Drug Administration, Silver Spring, Maryland, USA.
⁵ Laboratory of Pathology, Center for Cancer Research, NCI, NIH, Bethesda, Maryland, USA.
⁶ Department of Molecular Medicine, Mayo Clinic, Rochester, Minnesota, USA.
⁷ Animal Surgery and Resources Core, NHLBI, NIH, Bethesda, Maryland, USA.
⁸ iPSC Core, NHLBI, NIH, Bethesda, Maryland, USA.
⁹ Chemistry and Synthesis Center, NHLBI, NIH, Bethesda, Maryland, USA.
¹⁰ Laboratory of Cardiovascular Regenerative Medicine, NHLBI, NIH, Bethesda, Maryland, USA.

Abstract

Techniques that enable longitudinal tracking of cell fate after myocardial delivery are imperative for optimizing the efficacy of cell-based cardiac therapies. However, these approaches have been underutilized in preclinical models and clinical trials, and there is considerable demand for site-specific strategies achieving long-term expression of reporter genes compatible with safe noninvasive imaging. In this study, the rhesus sodium/iodide symporter (NIS) gene was incorporated into rhesus macaque induced pluripotent stem cells (RhiPSCs) via CRISPR/Cas9. Cardiomyocytes derived from NIS-RhiPSCs (NIS-RhiPSC-CMs) exhibited overall similar morphological and electrophysiological characteristics compared to parental control RhiPSC-CMs at baseline and with exposure to physiological levels of sodium iodide. Mice were injected intramyocardially with 2 million NIS-RhiPSC-CMs immediately following myocardial infarction, and serial positron emission tomography/computed tomography was performed with ¹⁸ F-tetrafluoroborate to monitor transplanted cells in vivo. NIS-RhiPSC-CMs could be detected until study conclusion at 8 to 10 weeks postinjection. This NIS-based molecular imaging platform, with optimal safety and sensitivity characteristics, is primed for translation into large-animal preclinical models and clinical trials.

Keywords: CRISPR/Cas9; NIS; PET; cardiomyocytes; electrophysiology; iPSCs; imaging; in vivo tracking; rhesus macaque; sodium iodide symporter.

Publication types

Research Support, N.I.H., Intramural

MeSH terms

Animals
CRISPR-Cas Systems / genetics*
Cell Differentiation
Disease Models, Animal
Humans
Induced Pluripotent Stem Cells / metabolism*
Mice
Myocytes, Cardiac / metabolism*
Symporters / genetics*
Transfection

Substances

Symporters
sodium-iodide symporter