Abstract
Cell reprogramming technology has allowed the in vitro control of cell fate transition, thus allowing for the generation of highly desired cell types to recapitulate in vivo developmental processes and architectures. However, the precise molecular mechanisms underlying the reprogramming process remain to be defined. Here, we show that depleting p53 and p21, which are barriers to reprogramming, yields a high reprogramming efficiency. Deletion of these factors results in a distinct mitochondrial background with low expression of oxidative phosphorylation subunits and mitochondrial fusion proteins, including mitofusin 1 and 2 (Mfn1/2). Importantly, Mfn1/2 depletion reciprocally inhibits the p53-p21 pathway and promotes both the conversion of somatic cells to a pluripotent state and the maintenance of pluripotency. Mfn1/2 depletion facilitates the glycolytic metabolic transition through the activation of the Ras-Raf and hypoxia-inducible factor 1α (HIF1α) signaling at an early stage of reprogramming. HIF1α is required for increased glycolysis and reprogramming by Mfn1/2 depletion. Taken together, these results demonstrate that Mfn1/2 constitutes a new barrier to reprogramming, and that Mfn1/2 ablation facilitates the induction of pluripotency through the restructuring of mitochondrial dynamics and bioenergetics.
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Abbreviations
- 2-DG:
-
2-deoxy-D-glucose
- AP:
-
alkaline phosphatase
- CM:
-
conditioned medium
- Drp1:
-
dynamin-related protein 1
- ESCs:
-
embryonic stem cells
- hESCs:
-
human ESCs
- HIF1α:
-
hypoxia-inducible factor 1α
- iPSC:
-
induced pluripotent stem cell
- KO:
-
knockout
- LDHA:
-
lactate dehydrogenase isoform A
- MACS:
-
magnetic-activated cell sorting
- MEFs:
-
mouse embryonic fibroblasts
- Mfn1/2:
-
mitofusin 1 and 2
- NEAA:
-
non-essential amino acids
- OSKM:
-
Oct4, Sox2, Klf4, and c-Myc
- OXPHOS:
-
oxidative phosphorylation
- PSCs:
-
pluripotent stem cells
- ROS:
-
reactive oxygen species
- RT:
-
room temperature
- UM:
-
unconditioned medium
- WT:
-
wild-type
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Acknowledgements
This work was supported by the National Research Foundation of Korea (NRF; 2010-020272(3), and 2012M3A9C7050224) and the KRIBB/NST research initiative program (NAP-09-3) grants funded by the Korea government (MSIP).
Author Contributions
Myung Jin Son: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript. Youjeong Kwon: collection and/or assembly of data, data analysis and interpretation. Mi-Young Son, Hoon-Sung Choi, and Binna Seol: collection and/or assembly of data. Seung-Wook Ryu: conception and design, data interpretation, final approval of manuscript. Chulhee Choi: conception and design, final approval of manuscript. Yee Sook Cho: financial support, data analysis and interpretation, manuscript writing, final approval of manuscript.
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Son, M., Kwon, Y., Son, MY. et al. Mitofusins deficiency elicits mitochondrial metabolic reprogramming to pluripotency. Cell Death Differ 22, 1957–1969 (2015). https://doi.org/10.1038/cdd.2015.43
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DOI: https://doi.org/10.1038/cdd.2015.43
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