C3G Regulates STAT3, ERK, Adhesion Signaling, and Is Essential for Differentiation of Embryonic Stem Cells

Abstract

SummaryC3G (RAPGEF1), engaged in multiple signaling pathways, is essential for the early development of the mouse. In this study, we have examined its role in mouse embryonic stem cell self-renewal and differentiation. C3G null cells generated by CRISPR mediated knock-in of a targeting vector exhibited enhanced clonogenicity and long-term self-renewal. They did not differentiate in response to LIF withdrawal when compared to the wild type ES cells and were defective for lineage commitment upon teratoma formation in vivo. Gene expression analysis of C3G KO cells showed misregulated expression of a large number of genes compared with WT cells. They express higher levels of self-renewal factors like KLF4 and ESRRB and show high STAT3 activity, and very low ERK activity compared to WT cells. Reintroduction of C3G expression in a KO line partially reverted expression of ESRRB, and KLF4, and ERK activity similar to that seen in WT cells. The expression of self-renewal factors was persistent for a longer time, and induction of lineage-specific markers was not seen when C3G KO cells were induced to form embryoid bodies. C3G KO cells showed poor adhesion and significantly reduced levels of pFAK, pPaxillin, and Integrin-β1, in addition to downregulation of the cluster of genes involved in cell adhesion, compared to WT cells. Our results show that C3G is essential for the regulation of STAT3, ERK, and adhesion signaling, to maintain pluripotency of mouse embryonic stem cells and enable their lineage commitment for differentiation. Graphical abstract

C3G down-regulation enhances pro-migratory and stemness properties of oval cells by promoting an epithelial-mesenchymal-like process

“…Thus, low levels of C3G in oval cells could still allow hepatocyte differentiation, while a total absence would not. On the other hand, the enhanced stemness of C3G knock-down hepatic progenitor/oval cells would agree with the above mentioned increased self-renewal found in C3G knock-out ESCs 36 .…”

“…On the other hand, it should be noticed that a reduced expression of C3G did not impair in vitro oval cell differentiation into hepatocytes in response to HGF, dexamethasone and oncostatin M in the presence of serum, even though HGF/MET signaling was defective. These results could partially contradict previous data showing that C3G promotes the differentiation of muscle cells 35 and megakaryocytes 20 , and with the fact that C3G knock-out in mouse embryonic stem cells (ESCs) impairs lineage commitment, while enhancing self-renewal and clonogenicity 36 . This potential discrepancy might be due to differences in the levels of C3G expression.…”

Early cellular and molecular signatures correlate with severity of West Nile virus infection

“…Strikingly, we found differential expression for 21 of 22 genes in at least one condition (p value <0.01, Wilcoxon rank-sum test) and highly significant differential expression for 16 of 22 (p value <0.001). Particularly interesting are 3 genes elevated in the acute samples that are significant across time and severity groups ( Figure 6 B): RAPGEF1 , a guanine nucleotide exchange factor involved in signal transmission and lineage-specific differentiation 83 ; LYN and REL , activating src family tyrosine kinases. In contrast, PPP3CA , a calcineurin protein which activates T cells, showed higher expression during convalescence, particularly in the symptomatic cases reflecting their higher inflammatory status.…”

LNC00115 Mediates Cisplatin Resistance by Regulating the miR-7/ERK Signalling Pathway in Ovarian Cancer

“…27 We predicted the targets of miR-7 using bioinformatics technology and determined the binding site of miR-7 on ERK by a dual-luciferase assay. The ERK/MAPK pathway regulates many important physiological and pathological processes in cells, such as cell growth, differentiation, 28 adaptation to environmental stress, and inflammation. 29 Because the ERK/MAPK pathway plays an important biological role and is highly activated in tumours, inhibition of this pathway is one of the most promising strategies for tumour targeted therapies.…”