論文

査読有り 国際誌
2019年12月20日

Overexpression of Na+/H+ exchanger 1 specifically induces cell death in human iPS cells via sustained activation of the Rho kinase ROCK.

The Journal of biological chemistry
  • Shigeo Wakabayashi
  • ,
  • Hirofumi Morihara
  • ,
  • Shunichi Yokoe
  • ,
  • Takatoshi Nakagawa
  • ,
  • Kazumasa Moriwaki
  • ,
  • Kiichiro Tomoda
  • ,
  • Michio Asahi

294
51
開始ページ
19577
終了ページ
19588
記述言語
英語
掲載種別
DOI
10.1074/jbc.RA119.010329

Understanding the specific properties of human induced pluripotent stem cells (iPSCs) is important for quality control of iPSCs. Having incidentally discovered that overexpression of plasma membrane Na+/H+ exchanger 1 (NHE1) induces cell death in iPSCs, we investigated the mechanism of NHE1-induced cell death. Doxycycline-induced NHE1 overexpression arrested cell growth, and nearly all cells were killed by a necrotic process within 72 h. NHE1 overexpression led to sustained activation of Rho-associated coiled-coil kinase (ROCK), accompanied by dramatic changes in cell shape, cell elongation, and swelling of peripheral cells in iPSC colonies, as well as marked stress fiber formation. The ROCK inhibitor Y27632 reduced NHE1-induced cell death. ROCK-dependent phenotypes were suppressed by a loss-of-function mutation of NHE1 and inhibited by an inhibitor of NHE1 activity, indicating that NHE1-mediated transport activity is required. Moreover, ROCK was activated by trimethylamine treatment-mediated cytosolic alkalinization and accumulated in the plasma membrane near NHE1 in peripheral iPSCs of cell colonies. By contrast, cell death did not occur in mesendoderm-like cells that had differentiated from iPSCs, indicating that the NHE1-mediated effects were specific for iPSCs. These results suggest that NHE1 overexpression specifically induces death of iPSCs via sustained ROCK activation, probably caused by an increase in local pH near NHE1. Finally, monensin, a Na+/H+ exchange ionophore, selectively killed iPSCs, suggesting that monensin could help eliminate iPSCs that remain after differentiation, a strategy that might be useful for improving regenerative medicine.

リンク情報
DOI
https://doi.org/10.1074/jbc.RA119.010329
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/31723030
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6926445
ID情報
  • DOI : 10.1074/jbc.RA119.010329
  • ISSN : 0021-9258
  • PubMed ID : 31723030
  • PubMed Central 記事ID : PMC6926445

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