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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Selection of iPSCs without mtDNA deletion for autologous cell therapy in a patient with Pearson syndrome

  • Yeonmi Lee (Department of Biomedical Science, College of Life Science, CHA University) ;
  • Jongsuk Han (Department of Biomedical Science, College of Life Science, CHA University) ;
  • Sae-Byeok Hwang (Department of Biomedical Science, College of Life Science, CHA University) ;
  • Soon-Suk Kang (Cell Therapy 3 Center, CHA Advanced Research Institute, CHA University) ;
  • Hyeoung-Bin Son (Department of Biomedical Science, College of Life Science, CHA University) ;
  • Chaeyeon Jin (Department of Biomedical Science, College of Life Science, CHA University) ;
  • Jae Eun Kim (Cell Therapy 3 Center, CHA Advanced Research Institute, CHA University) ;
  • Beom Hee Lee (Medical Genetics Center, Asan Medical Center, University of Ulsan College of Medicine) ;
  • Eunju Kang (Department of Biomedical Science, College of Life Science, CHA University)
  • Received : 2022.12.09
  • Accepted : 2023.05.02
  • Published : 2023.08.31
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Abstract

Screening for genetic defects in the cells should be examined for clinical application. The Pearson syndrome (PS) patient harbored nuclear mutations in the POLG and SSBP1 genes, which could induce systemic large-scale mitochondrial genome (mtDNA) deletion. We investigated iPSCs with mtDNA deletions in PS patient and whether deletion levels could be maintained during differentiation. The iPSC clones derived from skin fibroblasts (9% deletion) and blood mononuclear cells (24% deletion) were measured for mtDNA deletion levels. Of the 13 skin-derived iPSC clones, only 3 were found to be free of mtDNA deletions, whereas all blood-derived iPSC clones were found to be free of deletions. The iPSC clones with (27%) and without mtDNA deletion (0%) were selected and performed in vitro and in vivo differentiation, such as embryonic body (EB) and teratoma formation. After differentiation, the level of deletion was retained or increased in EBs (24%) or teratoma (45%) from deletion iPSC clone, while, the absence of deletions showed in all EBs and teratomas from deletion-free iPSC clones. These results demonstrated that non-deletion in iPSCs was maintained during in vitro and in vivo differentiation, even in the presence of nuclear mutations, suggesting that deletion-free iPSC clones could be candidates for autologous cell therapy in patients.

Keywords

Acknowledgement

This work was supported by the Ministry of Health and Welfare of Korea (HR16C0002(HI16C1559)), and intramural grants from the Asan Institute for Life Sciences, AMC (No. 2019-756).

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