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Spatially resolved cell polarity proteomics of a human epiblast model
Critical early steps in human embryonic development include polarization of the inner cell mass, followed by formation of an expanded lumen that will become the epiblast cavity. Recently described three-dimensional (3D) human pluripotent stem cell–derived cyst (hPSC-cyst) structures can replicate th...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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American Association for the Advancement of Science
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8064645/ https://www.ncbi.nlm.nih.gov/pubmed/33893097 http://dx.doi.org/10.1126/sciadv.abd8407 |
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author | Wang, Sicong Lin, Chien-Wei Carleton, Amber E. Cortez, Chari L. Johnson, Craig Taniguchi, Linnea E. Sekulovski, Nikola Townshend, Ryan F. Basrur, Venkatesha Nesvizhskii, Alexey I. Zou, Peng Fu, Jianping Gumucio, Deborah L. Duncan, Mara C. Taniguchi, Kenichiro |
author_facet | Wang, Sicong Lin, Chien-Wei Carleton, Amber E. Cortez, Chari L. Johnson, Craig Taniguchi, Linnea E. Sekulovski, Nikola Townshend, Ryan F. Basrur, Venkatesha Nesvizhskii, Alexey I. Zou, Peng Fu, Jianping Gumucio, Deborah L. Duncan, Mara C. Taniguchi, Kenichiro |
author_sort | Wang, Sicong |
collection | PubMed |
description | Critical early steps in human embryonic development include polarization of the inner cell mass, followed by formation of an expanded lumen that will become the epiblast cavity. Recently described three-dimensional (3D) human pluripotent stem cell–derived cyst (hPSC-cyst) structures can replicate these processes. To gain mechanistic insights into the poorly understood machinery involved in epiblast cavity formation, we interrogated the proteomes of apical and basolateral membrane territories in 3D human hPSC-cysts. APEX2-based proximity bioinylation, followed by quantitative mass spectrometry, revealed a variety of proteins without previous annotation to specific membrane subdomains. Functional experiments validated the requirement for several apically enriched proteins in cyst morphogenesis. In particular, we found a key role for the AP-1 clathrin adaptor complex in expanding the apical membrane domains during lumen establishment. These findings highlight the robust power of this proximity labeling approach for discovering novel regulators of epithelial morphogenesis in 3D stem cell–based models. |
format | Online Article Text |
id | pubmed-8064645 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Association for the Advancement of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-80646452021-05-05 Spatially resolved cell polarity proteomics of a human epiblast model Wang, Sicong Lin, Chien-Wei Carleton, Amber E. Cortez, Chari L. Johnson, Craig Taniguchi, Linnea E. Sekulovski, Nikola Townshend, Ryan F. Basrur, Venkatesha Nesvizhskii, Alexey I. Zou, Peng Fu, Jianping Gumucio, Deborah L. Duncan, Mara C. Taniguchi, Kenichiro Sci Adv Research Articles Critical early steps in human embryonic development include polarization of the inner cell mass, followed by formation of an expanded lumen that will become the epiblast cavity. Recently described three-dimensional (3D) human pluripotent stem cell–derived cyst (hPSC-cyst) structures can replicate these processes. To gain mechanistic insights into the poorly understood machinery involved in epiblast cavity formation, we interrogated the proteomes of apical and basolateral membrane territories in 3D human hPSC-cysts. APEX2-based proximity bioinylation, followed by quantitative mass spectrometry, revealed a variety of proteins without previous annotation to specific membrane subdomains. Functional experiments validated the requirement for several apically enriched proteins in cyst morphogenesis. In particular, we found a key role for the AP-1 clathrin adaptor complex in expanding the apical membrane domains during lumen establishment. These findings highlight the robust power of this proximity labeling approach for discovering novel regulators of epithelial morphogenesis in 3D stem cell–based models. American Association for the Advancement of Science 2021-04-23 /pmc/articles/PMC8064645/ /pubmed/33893097 http://dx.doi.org/10.1126/sciadv.abd8407 Text en Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC). https://creativecommons.org/licenses/by-nc/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license (https://creativecommons.org/licenses/by-nc/4.0/) , which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited. |
spellingShingle | Research Articles Wang, Sicong Lin, Chien-Wei Carleton, Amber E. Cortez, Chari L. Johnson, Craig Taniguchi, Linnea E. Sekulovski, Nikola Townshend, Ryan F. Basrur, Venkatesha Nesvizhskii, Alexey I. Zou, Peng Fu, Jianping Gumucio, Deborah L. Duncan, Mara C. Taniguchi, Kenichiro Spatially resolved cell polarity proteomics of a human epiblast model |
title | Spatially resolved cell polarity proteomics of a human epiblast model |
title_full | Spatially resolved cell polarity proteomics of a human epiblast model |
title_fullStr | Spatially resolved cell polarity proteomics of a human epiblast model |
title_full_unstemmed | Spatially resolved cell polarity proteomics of a human epiblast model |
title_short | Spatially resolved cell polarity proteomics of a human epiblast model |
title_sort | spatially resolved cell polarity proteomics of a human epiblast model |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8064645/ https://www.ncbi.nlm.nih.gov/pubmed/33893097 http://dx.doi.org/10.1126/sciadv.abd8407 |
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