Activation of STAT3-mediated ciliated cell survival protects against severe infection by respiratory syncytial virus
Caiqi Zhao, … , Paul H. Lerou, Xingbin Ai
Caiqi Zhao, … , Paul H. Lerou, Xingbin Ai
Published November 1, 2024
Citation Information: J Clin Invest. 2024;134(21):e183978. https://doi.org/10.1172/JCI183978.
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Activation of STAT3-mediated ciliated cell survival protects against severe infection by respiratory syncytial virus

Abstract

Respiratory syncytial virus (RSV) selectively targets ciliated cells in human bronchial epithelium and can cause bronchiolitis and pneumonia, mostly in infants. To identify molecular targets of intervention during RSV infection in infants, we investigated how age regulates RSV interaction with the bronchial epithelium barrier. Employing precision-cut lung slices and air-liquid interface cultures generated from infant and adult human donors, we found robust RSV virus spread and extensive apoptotic cell death only in infant bronchial epithelium. In contrast, adult bronchial epithelium showed no barrier damage and limited RSV infection. Single nuclear RNA-Seq revealed age-related insufficiency of an antiapoptotic STAT3 activation response to RSV infection in infant ciliated cells, which was exploited to facilitate virus spread via the extruded apoptotic ciliated cells carrying RSV. Activation of STAT3 and blockade of apoptosis rendered protection against severe RSV infection in infant bronchial epithelium. Lastly, apoptotic inhibitor treatment of a neonatal mouse model of RSV infection mitigated infection and inflammation in the lung. Taken together, our findings identify a STAT3-mediated antiapoptosis pathway as a target to battle severe RSV disease in infants.

Authors

Caiqi Zhao, Yan Bai, Wei Wang, Gaurang M. Amonkar, Hongmei Mou, Judith Olejnik, Adam J. Hume, Elke Mühlberger, Nicholas W. Lukacs, Rachel Fearns, Paul H. Lerou, Xingbin Ai

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Figure 4

Apoptotic cell extrusion in neonatal epithelium model following RSV infection can mediate virus spread.

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Apoptotic cell extrusion in neonatal epithelium model following RSV infe...
(A) Schematic of extrusion assays. Extruded cells were collected from apical washes of neonatal and adult ALI cultures at 2 and 4 dpi and analyzed in B–E. (B) Representative images of extruded cells on a cytometer (left panels) and double staining for RSV F protein (green) and ciliated cell markers (RFX3 and AceTUB [red])Blue color is nucleus staining and the overlay of red, green, and blue shows as pink color.) (right panels). (C) Quantification of the number of extruded cells. (D) Representative double staining for RSV F (green) protein and c-Casp-3 (red) (left panels) and quantification of the relative abundance of c-Casp-3+ cells (right panel). (E) Representative double staining for RSV F (green) protein and TUNEL(red) (left panels) and quantification of the relative abundance of TUNEL+ cells (right panel). (F) Schematic of infection assay by extruded cells. (G) Representative double staining for RSV F (green) and c-Casp-3 (red) after treatment with cell fraction and cell-free supernatant. Blue color is nucleus staining and the overlay of red, green, and blue shows as pink color. Each dot represents 1 donor. Bar graphs show mean ± SEM. ***P < 0.001 calculated by 2-way ANOVA followed by Dunn’s test in C. **P < 0.01 by 2-tailed Student’s t test in D and E. Scale bars: 5 μm (B) and 50 μm (D, E, and G).