Mechanical force sensing drives extrusion of Enterovirus A71-infected cells from colonic epithelial organoids

Abstract

Enterovirus A71 (EV-A71) causes severe disease upon systemic infection, sometimes leading to life-threatening neurological dysfunction. In most cases, infection is limited to the gastrointestinal tract, where virus is amplified for transmission. We used three-dimensional epithelial organoids generated from crypt stem cells of healthy patient colon tissue (colonoids) to investigate viral spread. Surprisingly, many infected cells were extruded from the apical surface of colonoids, leading to their removal from the epithelium. EV-A71 infected extruding cells were not frequently apoptotic and could propagate infection to uninfected monolayers and colonoids. Cell extrusion in healthy gastrointestinal epithelium is mediated either by apoptosis or cell-crowding forces sensed via the mechanosensitive ion channel Piezo-1. Treatment of infected colonoids with mechanosensitive ion-channel inhibitor GsMTx4 significantly reduced the infected cell extrusion. Instead, increased abundance of cell-free virus in media was observed. These results suggest a novel mechanism for extrusion of live, virus-infected cells through mechanical compression forces. In the gastrointestinal tract, apically extruded cells are released into the gut lumen and excreted in feces; therefore, extruded infected cells likely contribute to virus spread.

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The epithelium takes the stage in asthma and inflammatory bowel diseases

“…Conversely, crowding or compression arrests cell cycle and restores homeostatic cell numbers via cell extrusion, an evolutionarily conserved mechanism where a supracellular actomyosin cable formed around the unwanted cell ratchets in and down, resulting in seamless cell eviction without compromising barrier function (Rosenblatt et al, 2001;Eisenhoffer et al, 2012;McClatchey and Yap, 2012;Puliafito et al, 2012). In that sense, extrusion also works as an innate defence mechanism against external aggression, with healthy cells collectively squeezing cells infected by bacteria or viruses, thus limiting pathogen spreading in the monolayer and ensuring epithelial barrier function (Bastounis et al, 2021;Hippee et al, 2021;Lin et al, 2021;Moshiri et al, 2023).…”