Optimal ROS Signaling Is Critical for Nuclear Reprogramming.
Efficient nuclear reprogramming of somatic cells to pluripotency requires activation of innate immunity. Because innate immune activation triggers reactive oxygen species (ROS) signaling, we sought to determine whether there was a role of ROS signaling in nuclear reprogramming. We examined ROS production during the reprogramming of doxycycline (dox)-inducible mouse embryonic fibroblasts (MEFs) carrying the Yamanaka factors (Oct4, Sox2, Klf4, and c-Myc [OSKM]) into induced pluripotent stem cells (iPSCs). ROS generation was substantially increased with the onset of reprogramming. Depletion of ROS via antioxidants or Nox inhibitors substantially decreased reprogramming efficiency. Similarly, both knockdown and knockout of p22(phox)-a critical subunit of the Nox (1-4) complex-decreased reprogramming efficiency. However, excessive ROS generation using genetic and pharmacological approaches also impaired reprogramming. Overall, our data indicate that ROS signaling is activated early with nuclear reprogramming, and optimal levels of ROS signaling are essential to induce pluripotency.
Department of Cardiovascular Sciences, Center for Cardiovascular Regeneration, Houston Methodist Research Institute (HMRI), 6670 Bertner Avenue, Houston, TX 77030, USA.
Gang
G
Zhou
Department of Cardiovascular Sciences, Center for Cardiovascular Regeneration, Houston Methodist Research Institute (HMRI), 6670 Bertner Avenue, Houston, TX 77030, USA.
Shu
S
Meng
Department of Cardiovascular Sciences, Center for Cardiovascular Regeneration, Houston Methodist Research Institute (HMRI), 6670 Bertner Avenue, Houston, TX 77030, USA.
Yanhui
Y
Li
Department of Cardiovascular Sciences, Center for Cardiovascular Regeneration, Houston Methodist Research Institute (HMRI), 6670 Bertner Avenue, Houston, TX 77030, USA.
Yohannes T
YT
Ghebre
Department of Cardiovascular Sciences, Center for Cardiovascular Regeneration, Houston Methodist Research Institute (HMRI), 6670 Bertner Avenue, Houston, TX 77030, USA. Electronic address: jpcooke@houstonmethodist.org.
John P
JP
Cooke
D000818:Animals; D065150:Cellular Reprogramming; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction; D015854:Up-Regulation
C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D008562:Membrane Glycoproteins; D016328:NF-kappa B; D017382:Reactive Oxygen Species; C488842:Cybb protein, mouse; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases
CRISPR/Cas9; NADPH oxidase; Nrf2; iPSCs; nuclear reprogramming; reactive oxygen species