PMID- 32272203
OWN - NLM
STAT- MEDLINE
DCOM- 20201221
LR  - 20220727
IS  - 1872-8006 (Electronic)
IS  - 0304-4165 (Linking)
VI  - 1864
IP  - 8
DP  - 2020 Aug
TI  - Metabolic characterization of a paused-like pluripotent state.
PG  - 129612
LID - S0304-4165(20)30124-0 [pii]
LID - 10.1016/j.bbagen.2020.129612 [doi]
AB  - Embryonic diapause is a conserved reproductive strategy in which development 
      arrests at the blastocyst phase. Recently mammalian target of rapamycin (mTOR) 
      inhibition was shown to induce diapause on mouse blastocysts and a paused-like 
      state on mouse embryonic stem cells (mESCs). In this work, we aimed to further 
      characterize this new paused-pluripotent state, focusing on its glycolytic and 
      oxidative metabolic function. We therefore exposed mESCs, to the mTOR inhibitor 
      INK-128 and evaluated proliferation, pluripotency status and energy-related 
      metabolism, as well as the mTOR inhibition status and translational function. 
      Unexpectedly, in our hands INK-128 did not inhibit the phosphorylation of mTOR or 
      its downstream targets after 48 h. Accordingly, no alterations on protein 
      translational function were observed. Nonetheless, INK-128 could still 
      successfully induce a paused-like state in naive mESCs regardless of their 
      culturing conditions, by greatly slowing proliferation without affecting 
      pluripotency status. This effect was more prevalent in 2i cultured cells. 
      Interestingly, in this paused-like state, mESCs present a glucose-related 
      hypometabolic profile, which is a hallmark of diapaused blastocysts, with 
      decreased glycolytic and oxidative metabolism and decreased nutrient uptake. 
      Despite the lack of mTOR inhibition and translational suppression, INK-128 still 
      induced a paused-like pluripotent state through cell cycle and metabolic 
      modulation, rather than by translational suppression, suggesting more than one 
      avenue for this type of pluripotent phenotype.
CI  - Copyright (c) 2020 The Author(s). Published by Elsevier B.V. All rights reserved.
FAU - Sousa, Maria Ines
AU  - Sousa MI
AD  - CNC-Center for Neuroscience and Cell Biology, CIBB, Azinhaga de Santa Comba, Polo 
      3, University of Coimbra, Coimbra, Portugal; University of Coimbra, Department of 
      Life Sciences, Calcada Martim de Freitas, 3000-456 Coimbra, Portugal.
FAU - Correia, Bibiana
AU  - Correia B
AD  - CNC-Center for Neuroscience and Cell Biology, CIBB, Azinhaga de Santa Comba, Polo 
      3, University of Coimbra, Coimbra, Portugal.
FAU - Rodrigues, Ana Sofia
AU  - Rodrigues AS
AD  - CNC-Center for Neuroscience and Cell Biology, CIBB, Azinhaga de Santa Comba, Polo 
      3, University of Coimbra, Coimbra, Portugal. Electronic address: 
      sofiarodrigues@fmed.uc.pt.
FAU - Ramalho-Santos, Joao
AU  - Ramalho-Santos J
AD  - CNC-Center for Neuroscience and Cell Biology, CIBB, Azinhaga de Santa Comba, Polo 
      3, University of Coimbra, Coimbra, Portugal; University of Coimbra, Department of 
      Life Sciences, Calcada Martim de Freitas, 3000-456 Coimbra, Portugal. Electronic 
      address: jramalho@uc.pt.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200406
PL  - Netherlands
TA  - Biochim Biophys Acta Gen Subj
JT  - Biochimica et biophysica acta. General subjects
JID - 101731726
RN  - 0 (Benzoxazoles)
RN  - 0 (Pyrimidines)
RN  - EC 2.7.1.1 (mTOR protein, mouse)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
RN  - JGH0DF1U03 (sapanisertib)
SB  - IM
MH  - Animals
MH  - Benzoxazoles/pharmacology
MH  - Cell Cycle/drug effects
MH  - Cell Proliferation/drug effects
MH  - Cells, Cultured
MH  - Energy Metabolism
MH  - Mice
MH  - Pluripotent Stem Cells/*cytology/drug effects/*metabolism
MH  - Pyrimidines/pharmacology
MH  - TOR Serine-Threonine Kinases/metabolism
OTO - NOTNLM
OT  - Diapause
OT  - Embryonic stem cells
OT  - Metabolism
OT  - mTOR
COIS- Declaration of Competing Interest The authors declare that no conflict of 
      interest exists.
EDAT- 2020/04/10 06:00
MHDA- 2020/12/22 06:00
CRDT- 2020/04/10 06:00
PHST- 2019/10/08 00:00 [received]
PHST- 2020/03/02 00:00 [revised]
PHST- 2020/04/03 00:00 [accepted]
PHST- 2020/04/10 06:00 [pubmed]
PHST- 2020/12/22 06:00 [medline]
PHST- 2020/04/10 06:00 [entrez]
AID - S0304-4165(20)30124-0 [pii]
AID - 10.1016/j.bbagen.2020.129612 [doi]
PST - ppublish
SO  - Biochim Biophys Acta Gen Subj. 2020 Aug;1864(8):129612. doi: 
      10.1016/j.bbagen.2020.129612. Epub 2020 Apr 6.