Spatially resolved cell polarity proteomics of a human epiblast model. Sci Adv 2021 Apr;7(17)
Date
04/25/2021Pubmed ID
33893097Pubmed Central ID
PMC8064645DOI
10.1126/sciadv.abd8407Scopus ID
2-s2.0-85105042876 (requires institutional sign-in at Scopus site) 14 CitationsAbstract
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.
Author List
Wang S, Lin CW, Carleton AE, Cortez CL, Johnson C, Taniguchi LE, Sekulovski N, Townshend RF, Basrur V, Nesvizhskii AI, Zou P, Fu J, Gumucio DL, Duncan MC, Taniguchi KAuthors
Chien-Wei Lin PhD Associate Professor in the Data Science Institute department at Medical College of WisconsinKenichiro Taniguchi PhD Associate Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of Wisconsin
