Human Genomic Safe Harbors and the Suicide Gene-Based Safeguard System for iPSC-Based Cell Therapy

Author:

Kimura Yasuyoshi123,Shofuda Tomoko4,Higuchi Yuichiro5,Nagamori Ippei2,Oda Masaaki26,Nakamori Masayuki1,Onodera Masafumi7,Kanematsu Daisuke8,Yamamoto Atsuyo4,Katsuma Asako8,Suemizu Hiroshi5,Nakano Toru26,Kanemura Yonehiro8910,Mochizuki Hideki1

Affiliation:

1. Department of Neurology Graduate School of Medicine, Osaka University, Osaka, Japan

2. Department of Pathology Graduate School of Medicine, Osaka University, Osaka, Japan

3. Neuroregeneration and Stem Cell Programs Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

4. Division of Stem Cell Research, Department of Biomedical Research and Innovation Institute for Clinical Research, National Hospital Organization Osaka National Hospital, Osaka, Japan

5. Laboratory Animal Research Department, Biomedical Research Laboratory Central Institute for Experimental Animals, Kanagawa, Japan

6. Graduate School of Frontier Biosciences Osaka University, Osaka, Japan

7. Department of Human Genetics National Center for Child Health and Development, Tokyo, Japan

8. Division of Regenerative Medicine, Department of Biomedical Research and Innovation Institute for Clinical Research, National Hospital Organization Osaka National Hospital, Osaka, Japan

9. Department of Neurosurgery National Hospital Organization Osaka National Hospital, Osaka, Japan

10. Department of Physiology Keio University School of Medicine, Shinjuku, Tokyo, Japan

Abstract

Abstract The use of human induced pluripotent stem cells (hiPSCs) and recent advances in cell engineering have opened new prospects for cell-based therapy. However, there are concerns that must be addressed prior to their broad clinical applications and a major concern is tumorigenicity. Suicide gene approaches could eliminate wayward tumor-initiating cells even after cell transplantation, but their efficacy remains controversial. Another concern is the safety of genome editing. Our knowledge of human genomic safe harbors (GSHs) is still insufficient, making it difficult to predict the influence of gene integration on nearby genes. Here, we showed the topological architecture of human GSH candidates, AAVS1, CCR5, human ROSA26, and an extragenic GSH locus on chromosome 1 (Chr1-eGSH). Chr1-eGSH permitted robust transgene expression, but a 2 Mb-distant gene within the same topologically associated domain showed aberrant expression. Although knockin iPSCs carrying the suicide gene, herpes simplex virus thymidine kinase (HSV-TK), were sufficiently sensitive to ganciclovir in vitro, the resulting teratomas showed varying degrees of resistance to the drug in vivo. Our findings suggest that the Chr1-eGSH is not suitable for therapeutic gene integration and highlight that topological analysis could facilitate exploration of human GSHs for regenerative medicine applications. Our data indicate that the HSV-TK/ganciclovir suicide gene approach alone may be not an adequate safeguard against the risk of teratoma, and suggest that the combination of several distinct approaches could reduce the risks associated with cell therapy. Stem Cells Translational Medicine  2019;8:627&638

Funder

Japan Agency for Medical Research and Development, AMED

JSPS KAKENHI

Publisher

Oxford University Press (OUP)

Subject

Cell Biology,Developmental Biology,General Medicine

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