Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies

Overview

Journal iScience

Publisher Cell Press

Date 2019 Dec 11

PMID 31821946

Citations 13

Authors

Youjun Wu

Tammy Chang

Yan Long

He Huang

Fouad Kandeel

Jiing-Kuan Yee

Affiliations

Soon will be listed here.

Abstract

A major challenge in using human pluripotent stem cells (hPSCs) in therapy is the risk of teratoma formation due to contaminating undifferentiated stem cells. We used CRISPR-Cas9 for in-frame insertion of a suicide gene, iC9, into the endogenous SOX2 locus in human embryonic stem cell (ESC) line H1 for specific eradication of undifferentiated cells without affecting differentiated cells. This locus was chosen over NANOG and OCT4, two other well-characterized stem cell loci, due to significantly reduced off-target effect. We showed that undifferentiated H1-iC9 cells were induced to apoptosis by iC9 inducer AP1903, whereas differentiated cell lineages including hematopoietic cells, neurons, and islet beta-like cells were not affected. We also showed that AP1903 selectively removed undifferentiated H1-iC9 cells from a mixed cell population. This strategy therefore provides a layer of safety control before transplantation of a stem-cell-derived product in therapy.

Citing Articles

Strategies to Improve the Safety of iPSC-Derived β Cells for β Cell Replacement in Diabetes.

Pellegrini S, Zamarian V, Sordi V Transpl Int. 2022; 35:10575.

PMID: 36090777 PMC: 9448870. DOI: 10.3389/ti.2022.10575.

Importance of multiple endocrine cell types in islet organoids for type 1 diabetes treatment.

Heaton E, Jin S Transl Res. 2022; 250:68-83.

PMID: 35772687 PMC: 11554285. DOI: 10.1016/j.trsl.2022.06.014.

Tumorigenicity risk of iPSCs : nip it in the bud.

Zhong C, Liu M, Pan X, Zhu H Precis Clin Med. 2022; 5(1):pbac004.

PMID: 35692443 PMC: 9026204. DOI: 10.1093/pcmedi/pbac004.

Scalable manufacturing of clinical-grade differentiated cardiomyocytes derived from human-induced pluripotent stem cells for regenerative therapy.

Morita Y, Kishino Y, Fukuda K, Tohyama S Cell Prolif. 2022; 55(8):e13248.

PMID: 35534945 PMC: 9357358. DOI: 10.1111/cpr.13248.

Inducible caspase-9 suicide gene under control of endogenous oct4 to safeguard mouse and human pluripotent stem cell therapy.

Liu Y, Yang Y, Suo Y, Li C, Chen M, Zheng S Mol Ther Methods Clin Dev. 2022; 24:332-341.

PMID: 35229007 PMC: 8851157. DOI: 10.1016/j.omtm.2022.01.014.

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