An interplay of NOX1-derived ROS and oxygen determines the spermatogonial stem cell self-renewal efficiency under hypoxia
- Hiroko Morimoto1,
- Takuya Yamamoto2,3,4,5,
- Takehiro Miyazaki1,
- Narumi Ogonuki6,
- Atsuo Ogura6,
- Takashi Tanaka1,
- Mito Kanatsu-Shinohara1,
- Chihiro Yabe-Nishimura7,
- Hongliang Zhang8,
- Yves Pommier8,
- Andreas Trumpp9 and
- Takashi Shinohara1,3
- 1Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan;
- 2Department of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan;
- 3AMED-CREST, Chiyodaku, Tokyo 100-0004, Japan;
- 4Institute for the Advanced Study of Human Biology (WPI-ASHBi), Kyoto University, Kyoto 606-8501, Japan;
- 5Medical risk Avoidance Based on iPS Cells Team, RIKEN Center for Advanced Intelligence Project (AIP), Kyoto 606-8507, Japan;
- 6Bioresource Engineering Division, RIKEN BioResource Research Center, Ibaraki 305-0074, Japan;
- 7Department of Pharmacology, Kyoto Prefectural University of Medicine, Kyoto 606-8566, Japan;
- 8Developmental Therapeutics Branch and Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institute of Health, Bethesda, Maryland 20892, USA;
- 9Division of Stem Cells and Cancer, Deutsches Krebsforshungszentrum (DKFZ), 69120 Heidelberg, Germany
- Corresponding author: tshinoha{at}virus.kyoto-u.ac.jp
Abstract
Reactive oxygen species (ROS) produced by NADPH1 oxidase 1 (NOX1) are thought to drive spermatogonial stem cell (SSC) self-renewal through feed-forward production of ROS by the ROS-BCL6B-NOX1 pathway. Here we report the critical role of oxygen on ROS-induced self-renewal. Cultured SSCs proliferated poorly and lacked BCL6B expression under hypoxia despite increase in mitochondria-derived ROS. Due to lack of ROS amplification under hypoxia, NOX1-derived ROS were significantly reduced, and Nox1-deficient SSCs proliferated poorly under hypoxia but normally under normoxia. NOX1-derived ROS also influenced hypoxic response in vivo because Nox1-deficient undifferentiated spermatogonia showed significantly reduced expression of HIF1A, a master transcription factor for hypoxic response. Hypoxia-induced poor proliferation occurred despite activation of MYC and suppression of CDKN1A by HIF1A, whose deficiency exacerbated self-renewal efficiency. Impaired proliferation of Nox1- or Hif1a-deficient SSCs under hypoxia was rescued by Cdkn1a depletion. Consistent with these observations, Cdkn1a-deficient SSCs proliferated actively only under hypoxia but not under normoxia. On the other hand, chemical suppression of mitochondria-derived ROS or Top1mt mitochondria-specific topoisomerase deficiency did not influence SSC fate, suggesting that NOX1-derived ROS play a more important role in SSCs than mitochondria-derived ROS. These results underscore the importance of ROS origin and oxygen tension on SSC self-renewal.
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Footnotes
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Supplemental material is available for this article.
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Article published online ahead of print. Article and publication date are online at http://www.genesdev.org/cgi/doi/10.1101/gad.339903.120.
- Received May 22, 2020.
- Accepted December 18, 2020.
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