The stabilization of hypoxia inducible factor modulates
differentiation status and inhibits the proliferation of mouse
embryonic stem cells

J 2016

The stabilization of hypoxia inducible factor modulates differentiation status and inhibits the proliferation of mouse embryonic stem cells

BINÓ, Lucia, Jan KUČERA, Kateřina ŠTEFKOVÁ, Lenka ŠVIHÁLKOVÁ ŠINDLEROVÁ, Martina LÁNOVÁ et. al.

Basic information

Original name

The stabilization of hypoxia inducible factor modulates differentiation status and inhibits the proliferation of mouse embryonic stem cells

Authors

BINÓ, Lucia (703 Slovakia, belonging to the institution), Jan KUČERA (203 Czech Republic, belonging to the institution), Kateřina ŠTEFKOVÁ (203 Czech Republic, belonging to the institution), Lenka ŠVIHÁLKOVÁ ŠINDLEROVÁ (203 Czech Republic), Martina LÁNOVÁ (203 Czech Republic, belonging to the institution), Jana KUDOVÁ (203 Czech Republic, belonging to the institution), Lukáš KUBALA (203 Czech Republic, guarantor) and Jiří PACHERNÍK (203 Czech Republic, belonging to the institution)

Edition

Chemico-Biological Interactions, Clare, Elsevier Ireland Ltd. 2016, 0009-2797

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

Genetics and molecular biology

Country of publisher

Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 3.143

RIV identification code

RIV/00216224:14310/16:00089311

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1016/j.cbi.2015.12.007

UT WoS

000369152000022

Keywords in English

DMOG; Embryonic stem cells; HIF-1; Hypoxia; JNJ-42041935; Prolyl hydroxylase

Abstract

V originále

Hypoxic conditions are suggested to affect the differentiation status of stem cells (SC), including embryonic stem cells (ESC). Hypoxia inducible factor (HIF) is one of the main intracellular molecules responsible for the cellular response to hypoxia. Hypoxia stabilizes HIF by inhibiting the activity of HIF prolyl-hydroxylases (PHD), which are responsible for targeting HIF-alpha subunits for proteosomal degradation. To address the impact of HIF stabilization on the maintenance of the stemness signature of mouse ESC (mESC), we tested the influence of the inhibition of PHDs and hypoxia (1% O2 and 5% O2) on spontaneous ESC differentiation triggered by leukemia inhibitory factor withdrawal for 24 and 48 h. The widely used panhydroxylase inhibitor dimethyloxaloylglycine (DMOG) and PHD inhibitor JNJ-42041935 (JNJ) with suggested higher specificity towards PHDs were employed. Both inhibitors and both levels of hypoxia significantly increased HIF-1alpha and HIF-2alpha protein levels and HIF transcriptional activity in spontaneously differentiating mESC. This was accompanied by significant downregulation of cell proliferation manifested by the complete inhibition of DNA synthesis and partial arrest in the S phase after 48 h. Further, HIF stabilization enhanced downregulation of the expressions of some pluripotency markers (OCT-4, NANOG, ZFP-42, TNAP) in spontaneously differentiating mESC. However, at the same time, there was also a significant decrease in the expression of some genes selected as markers of cell differentiation (e.g. SOX1, BRACH T, ELF5). In conclusion, the short term stabilization of HIF mediated by the PHD inhibitors JNJ and DMOG and hypoxia did not prevent the spontaneous loss of pluripotency markers in mESC. However, it significantly downregulated the proliferation of these cells.

Citovat

BINÓ, Lucia, Jan KUČERA, Kateřina ŠTEFKOVÁ, Lenka ŠVIHÁLKOVÁ ŠINDLEROVÁ, Martina LÁNOVÁ, Jana KUDOVÁ, Lukáš KUBALA and Jiří PACHERNÍK. The stabilization of hypoxia inducible factor modulates differentiation status and inhibits the proliferation of mouse embryonic stem cells. Chemico-Biological Interactions. Clare: Elsevier Ireland Ltd., 2016, vol. 244, January, p. 204-214. ISSN 0009-2797. Available from: https://dx.doi.org/10.1016/j.cbi.2015.12.007.

@article{1333046,
   author = {Binó, Lucia and Kučera, Jan and Štefková, Kateřina and Švihálková Šindlerová, Lenka and Lánová, Martina and Kudová, Jana and Kubala, Lukáš and Pacherník, Jiří},
   article_location = {Clare},
   article_number = {January},
   doi = {http://dx.doi.org/10.1016/j.cbi.2015.12.007},
   keywords = {DMOG; Embryonic stem cells; HIF-1; Hypoxia; JNJ-42041935; Prolyl hydroxylase},
   language = {eng},
   issn = {0009-2797},
   journal = {Chemico-Biological Interactions},
   title = {The stabilization of hypoxia inducible factor modulates differentiation status and inhibits the proliferation of mouse embryonic stem cells},
   volume = {244},
   year = {2016}
}

TY  - JOUR
ID  - 1333046
AU  - Binó, Lucia - Kučera, Jan - Štefková, Kateřina - Švihálková Šindlerová, Lenka - Lánová, Martina - Kudová, Jana - Kubala, Lukáš - Pacherník, Jiří
PY  - 2016
TI  - The stabilization of hypoxia inducible factor modulates differentiation status and inhibits the proliferation of mouse embryonic stem cells
JF  - Chemico-Biological Interactions
VL  - 244
IS  - January
SP  - 204-214
EP  - 204-214
PB  - Elsevier Ireland Ltd.
SN  - 00092797
KW  - DMOG
KW  - Embryonic stem cells
KW  - HIF-1
KW  - Hypoxia
KW  - JNJ-42041935
KW  - Prolyl hydroxylase
N2  - Hypoxic conditions are suggested to affect the differentiation status of stem cells (SC), including embryonic stem cells (ESC). Hypoxia inducible factor (HIF) is one of the main intracellular molecules responsible for the cellular response to hypoxia. Hypoxia stabilizes HIF by inhibiting the activity of HIF prolyl-hydroxylases (PHD), which are responsible for targeting HIF-alpha subunits for proteosomal degradation. To address the impact of HIF stabilization on the maintenance of the stemness signature of mouse ESC (mESC), we tested the influence of the inhibition of PHDs and hypoxia (1% O2 and 5% O2) on spontaneous ESC differentiation triggered by leukemia inhibitory factor withdrawal for 24 and 48 h. The widely used panhydroxylase inhibitor dimethyloxaloylglycine (DMOG) and PHD inhibitor JNJ-42041935 (JNJ) with suggested higher specificity towards PHDs were employed. Both inhibitors and both levels of hypoxia significantly increased HIF-1alpha and HIF-2alpha protein levels and HIF transcriptional activity in spontaneously differentiating mESC. This was accompanied by significant downregulation of cell proliferation manifested by the complete inhibition of DNA synthesis and partial arrest in the S phase after 48 h. Further, HIF stabilization enhanced downregulation of the expressions of some pluripotency markers (OCT-4, NANOG, ZFP-42, TNAP) in spontaneously differentiating mESC. However, at the same time, there was also a significant decrease in the expression of some genes selected as markers of cell differentiation (e.g. SOX1, BRACH T, ELF5). In conclusion, the short term stabilization of HIF mediated by the PHD inhibitors JNJ and DMOG and hypoxia did not prevent the spontaneous loss of pluripotency markers in mESC. However, it significantly downregulated the proliferation of these cells.
ER  -

BINÓ, Lucia, Jan KUČERA, Kateřina ŠTEFKOVÁ, Lenka ŠVIHÁLKOVÁ ŠINDLEROVÁ, Martina LÁNOVÁ, Jana KUDOVÁ, Lukáš KUBALA and Jiří PACHERNÍK. The stabilization of hypoxia inducible factor modulates differentiation status and inhibits the proliferation of mouse embryonic stem cells. \textit{Chemico-Biological Interactions}. Clare: Elsevier Ireland Ltd., 2016, vol.~244, January, p.~204-214. ISSN~0009-2797. Available from: https://dx.doi.org/10.1016/j.cbi.2015.12.007.

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