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

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.

Cardiomyocyte Death and Genome-Edited Stem Cell Therapy for Ischemic Heart Disease

“…NIS is a transmembrane protein that enables imaging and identification of transplanted cells using radiotracers. When cardiomyocytes derived from these cells (NIS-RhiPSC-CM) were injected into MI-induced mice, these cells could be detected even 8-10 weeks after transplantation [133]. Although, further evaluation of cardiac function was not conducted in this research.…”

A brief review of reporter gene imaging in oncolytic virotherapy and gene therapy

“…Of particular interest, NIS has enabled tracking of OV, cell, and gene therapies in large animal models, the importance of which is discussed later. 30 , 61 , 70 , 71 , 72 , 73 , 74 , 75 , 76 The moderate sensitivity and excellent specificity of NIS in vivo imaging requires fewer animals for small and large animal studies, reducing animal use and costs and expediting pre-clinical and translational research.…”

“…To date, the only reporter gene to be repeatedly used for large animal deep-tissue imaging of OV or gene therapy is NIS, which has been studied in dogs, pigs, and non-human primates. 30 , 61 , 70 , 71 , 72 , 73 , 74 , 75 , 76 Use of the other genes discussed in this review for large animal imaging of OV and gene therapy is skewed heavily toward gene therapy and is sporadic. 87 , 152 , 153 , 154 , 155 , 156 , 157 , 158 , 159 , 160 , 161 The one exception is GFP, which has been used widely to test ocular gene therapies in dogs, cats, and non-human primates, because OI is possible due to the unique location and physiology of the eye.…”

The Advance of CRISPR-Cas9-Based and NIR/CRISPR-Cas9-Based Imaging System

“…This system successfully labeled the colon cancer cells ( Tasan et al, 2018 ). Moreover, the combination of PET and CRISPR-Cas9 had been applied in clinical translation of cell-based therapeutics ( Ostrominski et al, 2020 ). HSVtk gene was integrated into the AAVS1 locus of human urinary-induced pluripotent stem cell-derived cardiomyocytes (hUiCMs) using CRISPR-Cas9 system.…”