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Summary: Severe and often fatal opportunistic fungal infections arise frequently following mucosal damage caused by trauma or cytotoxic chemotherapy. Interaction of fungal pathogens with epithelial cells that comprise mucosae is a key early event associated with invasion, and, therefore, enhancing epithelial defense mechanisms may mitigate infection. Here, we establish a model of mold and yeast infection mediated by inducible epithelial cell loss in larval zebrafish. Epithelial cell loss by extrusion promotes exposure of laminin associated with increased fungal attachment, invasion, and larval lethality, whereas fungi defective in adherence or filamentation have reduced virulence. Transcriptional profiling identifies significant upregulation of the epidermal growth factor receptor ligand epigen (EPGN) upon mucosal damage. Treatment with recombinant human EPGN suppresses epithelial cell extrusion, leading to reduced fungal invasion and significantly enhanced survival. These data support the concept of augmenting epithelial restorative capacity to attenuate pathogenic invasion of fungi associated with human disease.

Cell injury poses a substantial challenge for epithelia homeostasis. Several cellular processes preserve epithelial barriers in response to apoptosis, but less is known about other forms of cell death, such as pyroptosis. Here we use an inducible caspase-1 system to analyze how colon epithelial monolayers respond to pyroptosis. We confirm that sporadic pyroptotic cells are physically eliminated from confluent monolayers by apical extrusion. This is accompanied by a transient defect in barrier function at the site of the pyroptotic cells. By visualizing cell shape changes and traction patterns in combination with cytoskeletal inhibitors, we show that rapid lamellipodial responses in the neighbor cells are responsible for correcting the leakage and resealing the barrier. Cell contractility is not required for this resealing response, in contrast to the response to apoptosis. Therefore, pyroptosis elicits a distinct homeostatic response from the epithelium that is driven by the stimulation of lamellipodia in neighbor cells. [Display omitted] • Caspase-1 activation induces lytic cell death and extrusion response in epithelia • Transient epithelial permeability occurs during extrusion of Casp-1-activated cells • Neighbor cells rapidly extend lamellipodia to re-establish epithelial homeostasis • Lamellipodia inhibition delays epithelial resealing during pyroptotic extrusion Bonfim-Melo et al. show that apical extrusion is the epithelial response to sporadic pyroptotic events. In contrast to apoptosis, pyroptosis causes a transient local increase in monolayer permeability that is corrected by rapid lamellipodial responses from the neighbors, rather than their contractility [ABSTRACT FROM AUTHOR]

Severe and often fatal opportunistic fungal infections arise frequently following mucosal damage caused by trauma or cytotoxic chemotherapy. Interaction of fungal pathogens with epithelial cells that comprise mucosae is a key early event associated with invasion, and, therefore, enhancing epithelial defense mechanisms may mitigate infection. Here, we establish a model of mold and yeast infection mediated by inducible epithelial cell loss in larval zebrafish. Epithelial cell loss by extrusion promotes exposure of laminin associated with increased fungal attachment, invasion, and larval lethality, whereas fungi defective in adherence or filamentation have reduced virulence. Transcriptional profiling identifies significant upregulation of the epidermal growth factor receptor ligand epigen (EPGN) upon mucosal damage. Treatment with recombinant human EPGN suppresses epithelial cell extrusion, leading to reduced fungal invasion and significantly enhanced survival. These data support the concept of augmenting epithelial restorative capacity to attenuate pathogenic invasion of fungi associated with human disease. [Display omitted] • Inducible epithelial cell loss allows modeling of fungal infection in zebrafish larvae • Extrusion of numerous cells exposes laminin and increases fungal adhesion and invasion • Cell extrusion stimulates epigen , mmp13a , and il1b expression and neutrophil recruitment • rhEPGN suppresses extrusion to provide protective effects against fungal invasion Wurster et al. show that extrusion of numerous epithelial cells from tissue can expose underlying extracellular matrix components to promote increased attachment and invasion of fungi associated with human disease. Treatment with recombinant human EPGN suppressed epithelial cell extrusion, leading to significantly reduced opportunistic fungal invasion. [ABSTRACT FROM AUTHOR]

Blood flow modulates endothelial cell (EC) response during angiogenesis. Shear stress is known to control gene expression related to the endothelial-mesenchymal transition and endothelial-hematopoietic transition. However, the impact of blood flow on the cellular processes associated with EC extrusion is less well understood. To address this question, we dynamically record EC movements and use 3D quantitative methods to segregate the contributions of various cellular processes to the cellular trajectories in the zebrafish dorsal aorta. We find that ECs spread toward the cell extrusion area following the tissue deformation direction dictated by flow-derived mechanical forces. Cell extrusion increases when blood flow is impaired. Similarly, the mechanosensor polycystic kidney disease 2 (pkd2) limits cell extrusion, suggesting that ECs actively sense mechanical forces in the process. These findings identify pkd2 and flow as critical regulators of EC extrusion and suggest that mechanical forces coordinate this process by maintaining ECs within the endothelium. • Endothelial cell (EC) nuclei are enriched in the ventral dorsal aorta (DA) during EC movement • DA wall deforms asymmetrically in response to pulsatile blood flow • Low blood flow or abnormal mechanosensing results in increased cell extrusion Campinho et al. show that cell extrusion associated with hematopoietic stem cell generation in the ventral dorsal aorta is under the control of blood flow-dependent tissue mechanics. The mechanosensitive channel trpp2 and blood flow limit endothelial cell extrusion originating from the endothelium during the process. [ABSTRACT FROM AUTHOR]