Cell-Surface Proteomics Identifies Differences in Signaling and Adhesion Protein Expression between Naive and Primed Human Pluripotent Stem Cells

• Published in: Stem cell reports 2020-05, Vol.14 (5), p.972-988
• Main Authors: Wojdyla, Katarzyna, Collier, Amanda J., Fabian, Charlene, Nisi, Paola S., Biggins, Laura, Oxley, David, Rugg-Gunn, Peter J.
• Format: Article
• Language: English
• Subjects: Cell Adhesion; Cell Adhesion Molecules - genetics; Cell Adhesion Molecules - metabolism; Cell Line; Cell Membrane - metabolism; cell state; cell-surface markers; embryonic; Folate Receptor 1 - genetics; Folate Receptor 1 - metabolism; human; Humans; Induced Pluripotent Stem Cells - metabolism; JAK-STAT; Membrane Glycoproteins - genetics; Membrane Glycoproteins - metabolism; plasma membrane; pluripotent stem cell; Proteome - genetics; Proteome - metabolism; proteomics; Receptors, Cytokine - genetics; Receptors, Cytokine - metabolism; reprogramming; Resource; Signal Transduction
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2020.03.017

Naive and primed human pluripotent stem cells (hPSC) provide valuable models to study cellular and molecular developmental processes. The lack of detailed information about cell-surface protein expression in these two pluripotent cell types prevents an understanding of how the cells communicate and interact with their microenvironments. Here, we used plasma membrane profiling to directly measure cell-surface protein expression in naive and primed hPSC. This unbiased approach quantified over 1,700 plasma membrane proteins, including those involved in cell adhesion, signaling, and cell interactions. Notably, multiple cytokine receptors upstream of JAK-STAT signaling were more abundant in naive hPSC. In addition, functional experiments showed that FOLR1 and SUSD2 proteins are highly expressed at the cell surface in naive hPSC but are not required to establish human naive pluripotency. This study provides a comprehensive stem cell proteomic resource that uncovers differences in signaling pathway activity and has identified new markers to define human pluripotent states. [Display omitted] •Direct measurement of plasma membrane proteins in two human pluripotent states•Proteomic resource contains quantitative analysis of signaling and adhesion proteins•Multiple cytokine receptors upstream of JAK-STAT pathway identified in naive cells•Functional experiments reveal FOLR1 and SUSD2 are not required for naive pluripotency Wojdyla et al. use plasma membrane profiling to directly measure cell-surface protein expression in naive and primed human pluripotent stem cells. This comprehensive proteomic resource quantifies shared and state-specific abundance of signaling, adhesion, and cell migration proteins, thereby identifying new cell-surface markers and leading to a better understanding of how pluripotent cells communicate and interact with their microenvironments.