TY - JOUR
T1 - The porcupine inhibitor WNT974 provokes ectodermal lineage differentiation of human embryonic stem cells
AU - Kalantary-Charvadeh, Ashkan
AU - Hosseini, Vahid
AU - Mehdizadeh, Amir
AU - Nazari Soltan Ahmad, Saeed
AU - Rahbarghazi, Reza
AU - Nozad Charoudeh, Hojjatollah
AU - Nouri, Mohammad
AU - Darabi, Masoud
N1 - Funding Information:
We acknowledge Dr. Yiu To Yeung at the MRC Centre of Reproductive Health, Queens Medical Research Institute, University of Edinburgh, UK, for insightful manuscript reviews and scientific discussions. The authors appreciate the support from Professor Mehdi Totonchi at the Royan Institute for Stem Cell Biology and Technology. The research was financially supported by a grant from the Stem Cell Research Center, Tabriz University of Medical Sciences (Grant Number: 65430) and the Iranian Council for Development of Stem Cell Sciences and Technologies. The last author was supported by an Alexander von Humboldt Research Award from Germany.
Funding Information:
We acknowledge Dr. Yiu To Yeung at the MRC Centre of Reproductive Health, Queens Medical Research Institute, University of Edinburgh, UK, for insightful manuscript reviews and scientific discussions. The authors appreciate the support from Professor Mehdi Totonchi at the Royan Institute for Stem Cell Biology and Technology. The research was financially supported by a grant from the Stem Cell Research Center, Tabriz University of Medical Sciences (Grant Number: 65430) and the Iranian Council for Development of Stem Cell Sciences and Technologies. The last author was supported by an Alexander von Humboldt Research Award from Germany.
Publisher Copyright:
© 2022 John Wiley & Sons Ltd.
PY - 2022/6
Y1 - 2022/6
N2 - Porcupine (Porcn) enzyme plays an essential role in Wnt signaling activation. Stearoyl-CoA desaturase-1 (SCD1) is required to provide Porcn substrates. The aim of this study was to determine the effect of a novel Porcn inhibitor on the fate of human embryonic stem cells (hESCs) and the reliance of Porcn on SCD1 activity. hESCs were cultured on a feeder layer or Matrigel-coated plates. Small molecules WNT974 (LGK-974) and CAY10566 were used to inhibit Porcn and SCD1 activity, respectively. We assessed the effect of Porcn inhibition on viability, expression of Wnt signaling targets, pluripotency markers, proliferation, differentiation, and protein fatty acylation. hESCs' conditioned medium (CM) containing secreted Wnt proteins were applied in rescue experiments. To examine the catalytic dependency of Porcn on SCD1, the results of combined inhibitor treatment were compared with the SCD1 inhibitor alone. LGK-974 at the selected concentrations showed mild effects on hESCs viability, but significantly reduced messenger RNA and protein expression of Wnt signaling targets (Axin-2 and c-Myc) and pluripotency markers (OCT-4 and SOX-2) (p <.05). Adding 1 μM of Porcn inhibitor reduced proliferation (p =.03) and enhanced differentiation capacity into ectodermal progenitors (p =.02), which were reverted by CM. Click chemistry reaction did not show significant alteration in protein fatty acylation upon LGK-974 treatment. Moreover, combined inhibitor treatment caused no further substantial reduction in Wnt signaling targets, pluripotency markers, and protein fatty acylation relative to CAY10566-treated cultures. The substrate availability for Porcn activity is regulated by SCD1 and targeting Porcn by LGK-974 prompts the transition of hESCs from self-renewal state to ectodermal lineage.
AB - Porcupine (Porcn) enzyme plays an essential role in Wnt signaling activation. Stearoyl-CoA desaturase-1 (SCD1) is required to provide Porcn substrates. The aim of this study was to determine the effect of a novel Porcn inhibitor on the fate of human embryonic stem cells (hESCs) and the reliance of Porcn on SCD1 activity. hESCs were cultured on a feeder layer or Matrigel-coated plates. Small molecules WNT974 (LGK-974) and CAY10566 were used to inhibit Porcn and SCD1 activity, respectively. We assessed the effect of Porcn inhibition on viability, expression of Wnt signaling targets, pluripotency markers, proliferation, differentiation, and protein fatty acylation. hESCs' conditioned medium (CM) containing secreted Wnt proteins were applied in rescue experiments. To examine the catalytic dependency of Porcn on SCD1, the results of combined inhibitor treatment were compared with the SCD1 inhibitor alone. LGK-974 at the selected concentrations showed mild effects on hESCs viability, but significantly reduced messenger RNA and protein expression of Wnt signaling targets (Axin-2 and c-Myc) and pluripotency markers (OCT-4 and SOX-2) (p <.05). Adding 1 μM of Porcn inhibitor reduced proliferation (p =.03) and enhanced differentiation capacity into ectodermal progenitors (p =.02), which were reverted by CM. Click chemistry reaction did not show significant alteration in protein fatty acylation upon LGK-974 treatment. Moreover, combined inhibitor treatment caused no further substantial reduction in Wnt signaling targets, pluripotency markers, and protein fatty acylation relative to CAY10566-treated cultures. The substrate availability for Porcn activity is regulated by SCD1 and targeting Porcn by LGK-974 prompts the transition of hESCs from self-renewal state to ectodermal lineage.
UR - http://www.scopus.com/inward/record.url?scp=85128382501&partnerID=8YFLogxK
U2 - 10.1002/cbf.3700
DO - 10.1002/cbf.3700
M3 - Journal articles
C2 - 35445405
AN - SCOPUS:85128382501
SN - 0263-6484
VL - 40
SP - 359
EP - 368
JO - Cell Biochemistry and Function
JF - Cell Biochemistry and Function
IS - 4
ER -
