BMB Reports

생화학분자생물학회 (Korean Society for Biochemistry and Molecular Biology)
권호 표지
DOI QR코드

DOI QR Code

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)
  • 투고 : 2022.12.09
  • 심사 : 2023.05.02
  • 발행 : 2023.08.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

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.

키워드

과제정보

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).

참고문헌

  1. Brand M, Orr A, Perevoshchikova I and Quinlan C (2013) The role of mitochondrial function and cellular bioenergetics in ageing and disease. Br J Dermatol 169, 1-8 https://doi.org/10.1111/bjd.12208
  2. Missiroli S, Genovese I, Perrone M, Vezzani B, Vitto VA and Giorgi C (2020) The role of mitochondria in inflammation: from cancer to neurodegenerative disorders. J Clin Med 9, 740
  3. Taanman JW (1999) The mitochondrial genome: structure, transcription, translation and replication. Biochim Biophys Acta 1410, 103-123 https://doi.org/10.1016/S0005-2728(98)00161-3
  4. Taylor RW and Turnbull DM (2005) Mitochondrial DNA mutations in human disease. Nat Rev Genet 6, 389-402 https://doi.org/10.1038/nrg1606
  5. Ryzhkova AI, Sazonova MA, Sinyov VV et al (2018) Mitochondrial diseases caused by mtDNA mutations: a minireview. Ther Clin Risk Manag. 14, 1933
  6. Greaves LC, Reeve AK, Taylor RW and Turnbull DM (2012) Mitochondrial DNA and disease. J Pathol 226, 274-286 https://doi.org/10.1002/path.3028
  7. DiMauro S and Schon EA (2003) Mitochondrial respiratory-chain diseases. N Engl J Med 348, 2656-2668 https://doi.org/10.1056/NEJMra022567
  8. Ferreira M, Evangelista T, Almeida LS et al (2011) Relative frequency of known causes of multiple mtDNA deletions: two novel POLG mutations. Neuromuscul Disord 21, 483-488 https://doi.org/10.1016/j.nmd.2011.03.011
  9. Fontana GA and Gahlon HL (2020) Mechanisms of replication and repair in mitochondrial DNA deletion formation. Nucleic Acids Res 48, 11244-11258 https://doi.org/10.1093/nar/gkaa804
  10. Rusecka J, Kaliszewska M, Bartnik E and Tonska K (2018) Nuclear genes involved in mitochondrial diseases caused by instability of mitochondrial DNA. J Appl Genet 59, 43-57 https://doi.org/10.1007/s13353-017-0424-3
  11. Korhonen JA, Pham XH, Pellegrini M and Falkenberg M (2004) Reconstitution of a minimal mtDNA replisome in vitro. EMBO J 23, 2423-2429 https://doi.org/10.1038/sj.emboj.7600257
  12. Hudson G, Amati-Bonneau P, Blakely EL et al (2008) Mutation of OPA1 causes dominant optic atrophy with external ophthalmoplegia, ataxia, deafness and multiple mitochondrial DNA deletions: a novel disorder of mtDNA maintenance. Brain 131, 329-337 https://doi.org/10.1093/brain/awm272
  13. Lee Y, Kim T, Lee M et al (2021) De novo development of mtDNA deletion due to decreased POLG and SSBP1 expression in humans. Genes 12, 284
  14. Kang E, Wang X, Tippner-Hedges R et al (2016) Age-related accumulation of somatic mitochondrial DNA mutations in adult-derived human iPSCs. Cell Stem Cell 18, 625-636 https://doi.org/10.1016/j.stem.2016.02.005
  15. Ma H, Folmes CD, Wu J et al (2015) Metabolic rescue in pluripotent cells from patients with mtDNA disease. Nature 524, 234-238 https://doi.org/10.1038/nature14546
  16. Park J, Lee Y, Shin J et al (2019) Mitochondrial genome mutations in mesenchymal stem cells derived from human dental induced pluripotent stem cells. BMB Rep 52, 689
  17. Cherry AB, Gagne KE, Mcloughlin EM et al (2013) Induced pluripotent stem cells with a mitochondrial DNA deletion. Stem Cells 31, 1287-1297 https://doi.org/10.1002/stem.1354
  18. Van Tienen F, Zelissen R, Timmer E et al (2019) Healthy, mtDNA-mutation free mesoangioblasts from mtDNA patients qualify for autologous therapy. Stem Cell Res Ther 10, 1-10 https://doi.org/10.1186/s13287-019-1510-8
  19. Bang JS, Choi NY, Lee M, Ko K, Park YS and Ko K (2019) Reprogramming of cancer cells into induced pluripotent stem cells questioned. Int J Stem Cells 12, 430-439 https://doi.org/10.15283/ijsc19067
  20. Chen T, He J, Shen L et al (2011) The mitochondrial DNA 4,977-bp deletion and its implication in copy number alteration in colorectal cancer. BMC Med Genet 12, 1-9 https://doi.org/10.1186/1471-2350-12-8
  21. Lee HJ, Choi NY, Lee SW et al (2019) Alteration of genomic imprinting status of human parthenogenetic induced pluripotent stem cells during neural lineage differentiation. Int J Stem Cells 12, 31-42 https://doi.org/10.15283/ijsc18084
  22. So S, Lee Y, Choi J et al (2020) The Rho-associated kinase inhibitor fasudil can replace Y-27632 for use in human pluripotent stem cell research. PLoS One 15, e0233057
  23. Kwon J, Lee N, Jeon I et al (2012) Neuronal differentiation of a human induced pluripotent stem cell line (FS-1) derived from newborn foreskin fibroblasts. Int J Stem Cells 5, 140
  24. Malhotra N (2016) Induced pluripotent stem (iPS) cells in dentistry: a review. Int J Stem Cells 9, 176-185 https://doi.org/10.15283/ijsc16029
  25. Madrid M, Sumen C, Aivio S and Saklayen N (2021) Autologous induced pluripotent stem cell-based cell therapies: promise, progress, and challenges. Curr Protoc 1, e88
  26. Abi Chahine N, Wehbe T, Rashed J, Hilal R and Elias N (2016) Autologous bone marrow derived stem cells for the treatment of multiple sclerosis. Int J Stem Cells 9, 207-212 https://doi.org/10.15283/ijsc16049
  27. Kazmi B, Inglefield CJ and Lewis MP (2009) Autologous cell therapy: current treatments and future prospects. Wounds 21, 234-242
  28. Li C, Zhao H, Cheng L and Wang B (2021) Allogeneic vs. autologous mesenchymal stem/stromal cells in their medication practice. Cell Biosci 11, 1-21 https://doi.org/10.1186/s13578-020-00515-y
  29. Mohamed SA, Wesch D and Blumenthal A (2004) Detection of the 4977 bp deletion of mitochondrial DNA in different human blood cells. Exp Gerontol 39, 181-188 https://doi.org/10.1016/j.exger.2003.10.011
  30. Isogai S, Yamamoto N, Hiramatsu N et al (2018) Preparation of induced pluripotent stem cells using human peripheral blood monocytes. Cell Reprogram 20, 347-355 https://doi.org/10.1089/cell.2018.0024
  31. Russell OM, Fruh I, Rai PK et al (2018) Preferential amplification of a human mitochondrial DNA deletion in vitro and in vivo. Sci Rep 8, 1-10 https://doi.org/10.1038/s41598-018-20064-2
  32. Phillips NR, Sprouse ML and Roby RK (2014) Simultaneous quantification of mitochondrial DNA copy number and deletion ratio: a multiplex real-time PCR assay. Sci Rep 4, 1-7