Cargando…

NODAL/TGFβ signalling mediates the self-sustained stemness induced by PIK3CA(H1047R) homozygosity in pluripotent stem cells

Activating PIK3CA mutations are known ‘drivers’ of human cancer and developmental overgrowth syndromes. We recently demonstrated that the ‘hotspot’ PIK3CA(H1047R) variant exerts unexpected allele dose-dependent effects on stemness in human pluripotent stem cells (hPSCs). In this study, we combine hi...

Descripción completa

Detalles Bibliográficos
Autores principales: Madsen, Ralitsa R., Longden, James, Knox, Rachel G., Robin, Xavier, Völlmy, Franziska, Macleod, Kenneth G., Moniz, Larissa S., Carragher, Neil O., Linding, Rune, Vanhaesebroeck, Bart, Semple, Robert K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Company of Biologists Ltd 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7969366/
https://www.ncbi.nlm.nih.gov/pubmed/33514588
http://dx.doi.org/10.1242/dmm.048298
_version_ 1783666229184036864
author Madsen, Ralitsa R.
Longden, James
Knox, Rachel G.
Robin, Xavier
Völlmy, Franziska
Macleod, Kenneth G.
Moniz, Larissa S.
Carragher, Neil O.
Linding, Rune
Vanhaesebroeck, Bart
Semple, Robert K.
author_facet Madsen, Ralitsa R.
Longden, James
Knox, Rachel G.
Robin, Xavier
Völlmy, Franziska
Macleod, Kenneth G.
Moniz, Larissa S.
Carragher, Neil O.
Linding, Rune
Vanhaesebroeck, Bart
Semple, Robert K.
author_sort Madsen, Ralitsa R.
collection PubMed
description Activating PIK3CA mutations are known ‘drivers’ of human cancer and developmental overgrowth syndromes. We recently demonstrated that the ‘hotspot’ PIK3CA(H1047R) variant exerts unexpected allele dose-dependent effects on stemness in human pluripotent stem cells (hPSCs). In this study, we combine high-depth transcriptomics, total proteomics and reverse-phase protein arrays to reveal potentially disease-related alterations in heterozygous cells, and to assess the contribution of activated TGFβ signalling to the stemness phenotype of homozygous PIK3CA(H1047R) cells. We demonstrate signalling rewiring as a function of oncogenic PI3K signalling strength, and provide experimental evidence that self-sustained stemness is causally related to enhanced autocrine NODAL/TGFβ signalling. A significant transcriptomic signature of TGFβ pathway activation in heterozygous PIK3CA(H1047R) was observed but was modest and was not associated with the stemness phenotype seen in homozygous mutants. Notably, the stemness gene expression in homozygous PIK3CA(H1047R) hPSCs was reversed by pharmacological inhibition of NODAL/TGFβ signalling, but not by pharmacological PI3Kα pathway inhibition. Altogether, this provides the first in-depth analysis of PI3K signalling in hPSCs and directly links strong PI3K activation to developmental NODAL/TGFβ signalling. This work illustrates the importance of allele dosage and expression when artificial systems are used to model human genetic disease caused by activating PIK3CA mutations. This article has an associated First Person interview with the first author of the paper.
format Online
Article
Text
id pubmed-7969366
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher The Company of Biologists Ltd
record_format MEDLINE/PubMed
spelling pubmed-79693662021-03-18 NODAL/TGFβ signalling mediates the self-sustained stemness induced by PIK3CA(H1047R) homozygosity in pluripotent stem cells Madsen, Ralitsa R. Longden, James Knox, Rachel G. Robin, Xavier Völlmy, Franziska Macleod, Kenneth G. Moniz, Larissa S. Carragher, Neil O. Linding, Rune Vanhaesebroeck, Bart Semple, Robert K. Dis Model Mech Research Article Activating PIK3CA mutations are known ‘drivers’ of human cancer and developmental overgrowth syndromes. We recently demonstrated that the ‘hotspot’ PIK3CA(H1047R) variant exerts unexpected allele dose-dependent effects on stemness in human pluripotent stem cells (hPSCs). In this study, we combine high-depth transcriptomics, total proteomics and reverse-phase protein arrays to reveal potentially disease-related alterations in heterozygous cells, and to assess the contribution of activated TGFβ signalling to the stemness phenotype of homozygous PIK3CA(H1047R) cells. We demonstrate signalling rewiring as a function of oncogenic PI3K signalling strength, and provide experimental evidence that self-sustained stemness is causally related to enhanced autocrine NODAL/TGFβ signalling. A significant transcriptomic signature of TGFβ pathway activation in heterozygous PIK3CA(H1047R) was observed but was modest and was not associated with the stemness phenotype seen in homozygous mutants. Notably, the stemness gene expression in homozygous PIK3CA(H1047R) hPSCs was reversed by pharmacological inhibition of NODAL/TGFβ signalling, but not by pharmacological PI3Kα pathway inhibition. Altogether, this provides the first in-depth analysis of PI3K signalling in hPSCs and directly links strong PI3K activation to developmental NODAL/TGFβ signalling. This work illustrates the importance of allele dosage and expression when artificial systems are used to model human genetic disease caused by activating PIK3CA mutations. This article has an associated First Person interview with the first author of the paper. The Company of Biologists Ltd 2021-03-11 /pmc/articles/PMC7969366/ /pubmed/33514588 http://dx.doi.org/10.1242/dmm.048298 Text en © 2021. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/4.0This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
spellingShingle Research Article
Madsen, Ralitsa R.
Longden, James
Knox, Rachel G.
Robin, Xavier
Völlmy, Franziska
Macleod, Kenneth G.
Moniz, Larissa S.
Carragher, Neil O.
Linding, Rune
Vanhaesebroeck, Bart
Semple, Robert K.
NODAL/TGFβ signalling mediates the self-sustained stemness induced by PIK3CA(H1047R) homozygosity in pluripotent stem cells
title NODAL/TGFβ signalling mediates the self-sustained stemness induced by PIK3CA(H1047R) homozygosity in pluripotent stem cells
title_full NODAL/TGFβ signalling mediates the self-sustained stemness induced by PIK3CA(H1047R) homozygosity in pluripotent stem cells
title_fullStr NODAL/TGFβ signalling mediates the self-sustained stemness induced by PIK3CA(H1047R) homozygosity in pluripotent stem cells
title_full_unstemmed NODAL/TGFβ signalling mediates the self-sustained stemness induced by PIK3CA(H1047R) homozygosity in pluripotent stem cells
title_short NODAL/TGFβ signalling mediates the self-sustained stemness induced by PIK3CA(H1047R) homozygosity in pluripotent stem cells
title_sort nodal/tgfβ signalling mediates the self-sustained stemness induced by pik3ca(h1047r) homozygosity in pluripotent stem cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7969366/
https://www.ncbi.nlm.nih.gov/pubmed/33514588
http://dx.doi.org/10.1242/dmm.048298
work_keys_str_mv AT madsenralitsar nodaltgfbsignallingmediatestheselfsustainedstemnessinducedbypik3cah1047rhomozygosityinpluripotentstemcells
AT longdenjames nodaltgfbsignallingmediatestheselfsustainedstemnessinducedbypik3cah1047rhomozygosityinpluripotentstemcells
AT knoxrachelg nodaltgfbsignallingmediatestheselfsustainedstemnessinducedbypik3cah1047rhomozygosityinpluripotentstemcells
AT robinxavier nodaltgfbsignallingmediatestheselfsustainedstemnessinducedbypik3cah1047rhomozygosityinpluripotentstemcells
AT vollmyfranziska nodaltgfbsignallingmediatestheselfsustainedstemnessinducedbypik3cah1047rhomozygosityinpluripotentstemcells
AT macleodkennethg nodaltgfbsignallingmediatestheselfsustainedstemnessinducedbypik3cah1047rhomozygosityinpluripotentstemcells
AT monizlarissas nodaltgfbsignallingmediatestheselfsustainedstemnessinducedbypik3cah1047rhomozygosityinpluripotentstemcells
AT carragherneilo nodaltgfbsignallingmediatestheselfsustainedstemnessinducedbypik3cah1047rhomozygosityinpluripotentstemcells
AT lindingrune nodaltgfbsignallingmediatestheselfsustainedstemnessinducedbypik3cah1047rhomozygosityinpluripotentstemcells
AT vanhaesebroeckbart nodaltgfbsignallingmediatestheselfsustainedstemnessinducedbypik3cah1047rhomozygosityinpluripotentstemcells
AT semplerobertk nodaltgfbsignallingmediatestheselfsustainedstemnessinducedbypik3cah1047rhomozygosityinpluripotentstemcells