A1 Refereed original research article in a scientific journal
De Novo Gene Transcription of Connexin Mediates Cytoplasmic Fluid Exchange and Flocking Transitions in Physiological and Cancerous Epithelial Systems
Authors: Abdo, Hind; Barzaghi, Leonardo; Shen, Yuan; Bellini, Edoardo; Martini, Emanuele; Magni, Serena; Barozzi, Sara; Orsenigo, Fabrizio; Parazzoli, Dario; Beznoussenko, Galina V.; Franco, Jasmin Di; Krautgasser, Fabian; Kaivola, Jasmin; Cinquanta, Mario; Lazzarin, Alessandro; Sigismund, Sara; Ivaska, Johanna; Cerbino, Roberto; Scita, Giorgio
Publisher: Wiley
Publication year: 2025
Journal: Advanced Science
Article number: e08648
eISSN: 2198-3844
DOI: https://doi.org/10.1002/advs.202508648
Publication's open availability at the time of reporting: Open Access
Publication channel's open availability : Open Access publication channel
Web address : https://doi.org/10.1002/advs.202508648
Self-archived copy’s web address: https://research.utu.fi/converis/portal/detail/Publication/508253609
Self-archived copy's licence: CC BY
Self-archived copy's version: Publisher`s PDF
The initial invasion of tumors requires a transition from a solid, jammed state to a fluid-like, flocking, unjammed state that enables collective migration. Here, we show that de novo gene transcription is essential for the emergence of flocking in epithelial tissues and identify connexins (Cx) as key mediators of this transition. Using quiescent HaCaT keratinocytes, tumorigenic A431 epidermoid carcinoma cells, primary bronchial epithelial explants, and vocal fold carcinoma (VFC) cells, we find that flocking induction depends on transcriptional programs activated downstream of epidermal growth factor (EGF). EGF stimulation upregulates Cx26 and Cx31 and enhances gap-junctional intercellular communication (GJIC), which is necessary-though not sufficient-to generate the large-scale cell-volume fluctuations and density heterogeneity that accompany unjamming. Sustained signaling through extracellular signal-regulated kinase 1/2 (ERK1/2) and AKT serine/threonine kinase (AKT) downstream of the EGF receptor (EGFR) is required for connexin induction, linking mechanical state transitions to extracellular cues. Pharmacological inhibition and CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR associated protein 9) knockout of connexins block unjamming and collective motility. VFC cells display constitutively elevated connexins and persistent flocking that is highly sensitive to connexin inhibition. Consistently, high Cx26 expression correlates with reduced survival across carcinomas. These findings reveal a transcriptionally controlled, connexin-dependent mechanism that enables tissue fluidization and collective invasion.
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Funding information in the publication:
The authors thank V. Dall'Olio and L. Tizzoni at Cogentech for qRT-PCR services. The work was supported by ERC-Synergy (Grant# 801101071470), AIRC-IG (Grant#22821), AIRC 5X1000 (#22759), the Italian Ministry of University and Research (PRIN202223GSCIT_01/G53D23002570006/20229RM8A_001; COMBINE/G53D23007040001/P2022RH4HH002; PNRR_CN3RNA_SPOKE/G43C22001320007) to G.S.; AIRC IG 2022-ID 27101 to N.G.; ERC-CoG2020 #101002280, AIRC IG #24415, the Italian Ministry of University and Scientific Research, PRIN 2022, prot.2022W93FTW and the Italian Ministry of Health (Ricerca Corrente 2023-2024 and 5 per 1000 funds) to S.S.; Fondazione Veronesi Postdoctoral fellowship 2021-2024 to H.A.; J.K. is supported by the University of Turku Doctoral Program for Molecular Medicine and the Finnish Cultural Foundation.
