Sox2 controls neural stem cell self‐renewal through a Fos‐centered gene regulatory network. (29th March 2021)

• Record Type: Journal Article
• Authors: Pagin, Miriam; Pernebrink, Mattias; Giubbolini, Simone; Barone, Cristiana; Sambruni, Gaia; Zhu, Yanfen; Chiara, Matteo; Ottolenghi, Sergio; Pavesi, Giulio; Wei, Chia‐Lin; Cantù, Claudio; Nicolis, Silvia K.
• Is Part Of: Stem cells. Volume 39:Number 8(2021)
• Page Start: 1107
• Page End: 1119
• Publication Date: 2021-03-29
• Subjects: AP1 inhibitor T‐5224 -- CRISPR -- CUT&RUN -- Fos -- lentiviral vector -- neural stem cells (NSCs) -- self‐renewal -- Socs3 -- Sox2 -- transcription factors; Cloning -- Periodicals; Clone cells -- Periodicals; Stem cells -- Periodicals; Cell Differentiation -- Periodicals; Cell Division -- Periodicals; Clone Cells -- Periodicals; Hematopoietic Stem Cells -- Periodicals; Stem Cells -- Periodicals; 571.84
• DOI: 10.1002/stem.3373
• Languages: English
• ISSNs: 1066-5099

Abstract

Abstract: The Sox2 transcription factor is necessary for the long‐term self‐renewal of neural stem cells (NSCs). Its mechanism of action is still poorly defined. To identify molecules regulated by Sox2, and acting in mouse NSC maintenance, we transduced, into Sox2‐deleted NSC, genes whose expression is strongly downregulated following Sox2 loss (Fos, Jun, Egr2), individually or in combination. Fos alone rescued long‐term proliferation, as shown by in vitro cell growth and clonal analysis. Furthermore, pharmacological inhibition by T‐5224 of FOS/JUN AP1 complex binding to its targets decreased cell proliferation and expression of the putative target Suppressor of cytokine signaling 3 (Socs3). Additionally, Fos requirement for efficient long‐term proliferation was demonstrated by the reduction of NSC clones capable of long‐term expansion following CRISPR/Cas9‐mediated Fos inactivation. Previous work showed that the Socs3 gene is strongly downregulated following Sox2 deletion, and its re‐expression by lentiviral transduction rescues long‐term NSC proliferation. Fos appears to be an upstream regulator of Socs3, possibly together with Jun and Egr2; indeed, Sox2 re‐expression in Sox2‐deleted NSC progressively activates both Fos and Socs3 expression; in turn, Fos transduction activates Socs3 expression. Based on available SOX2 ChIPseq and ChIA‐PET data, we propose a model whereby Sox2 is a direct activator of both Socs3 and Fos, as well as possibly Jun and Egr2; furthermore, we provide direct evidence for FOS and JUN binding on Socs3 promoter, suggesting direct transcriptional regulation. These results provide the basis for developing a model of a network of interactions, regulating critical effectors of NSC proliferation and long‐term maintenance. Abstract : Sox2 is necessary for long‐term neural stem cell (NSC) self‐renewal and for Socs3, Fos, Jun, and Egr2 gene expression. We previously showed that Socs3 overexpression rescues Sox2‐deleted NSC self‐renewal. Fos, Jun, and Egr2 transduction into Sox2‐deleted NSC rescues self‐renewal, Fos alone being both sufficient and necessary. Sox2 transduction upregulates Fos and Socs3 in Sox2‐deleted cells, and Fos upregulates Socs3, defining a Sox2‐dependent regulatory network.