Development of selective cytotoxic viral vectors for concentration of undifferentiated cells in cardiomyocytes derived from human induced pluripotent stem cells

  • Sci Rep. 2019 Mar 6;9(1):3630. doi: 10.1038/s41598-018-36848-5.
Ken Kono  1 ,  Rumi Sawada  1 ,  Takuya Kuroda  1 ,  Satoshi Yasuda  1 ,  Satoko Matsuyama  1  2 ,  Akifumi Matsuyama  3 ,  Hiroyuki Mizuguchi  4 ,  Yoji Sato  5  6  7  8
Affiliations
  • 1. Division of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kanagawa, Japan.
  • 2. Platform of Therapeutics for Rare Disease, National Institutes of Biomedical Innovation, Health and Nutrition, Hyogo, Japan.
  • 3. Department of Regenerative Medicine, School of Medicine, Fujita Health University, Aichi, Japan.
  • 4. Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
  • 5. Division of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kanagawa, Japan. [email protected].
  • 6. Department of Quality Assurance Science for Pharmaceuticals, Graduate School of Pharmaceutical Sciences, Nagoya City University, Aichi, Japan. [email protected].
  • 7. Department of Cellular and Gene Therapy Products, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan. [email protected].
  • 8. Department of Translational Pharmaceutical Sciences, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan. [email protected].
PMID: 30842516 DOI: 10.1038/s41598-018-36848-5
Abstract

Cell-processed therapeutic products (CTPs) derived from human pluripotent stem cells (hPSCs) have innovative applications in regenerative medicine. However, undifferentiated hPSCs possess tumorigenic potential; thus, sensitive methods for the detection of residual undifferentiated hPSCs are essential for the clinical use of hPSC-derived CTPs. The detection limit of the methods currently available is 1/105 (0.001%, undifferentiated hPSCs/differentiated cells) or more, which could be insufficient for the detection of residual hPSCs when CTPs contain more than 1 × 105 cells. In this study, we developed a novel approach to overcome this challenge, using adenovirus and adeno-associated virus (AdV and AAV)-based selective cytotoxic vectors. We constructed AdV and AAV vectors that possess a suicide gene, iCaspase 9 (iCasp9), regulated by the CMV promoter, which is dormant in hPSCs, for the selective expression of iCasp9 in differentiated cells. As expected, AdV/CMV-iCasp9 and AAV/CMV-iCasp9 exhibited cytotoxicity in cardiomyocytes but not in human induced pluripotent stem cells (hiPSCs). The vectors also induced Apoptosis in hiPSC-derived cardiomyocytes, and the surviving cells exhibited higher levels of hPSC marker expression. These results indicate that the AdV- and AAV-based cytotoxic vectors concentrate cells expressing the undifferentiated cell markers in hiPSC-derived products and are promising biological tools for verifying the quality of CTPs.

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