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Unbiased proteomics following inflammasome activation identifies caspase targets in primary intestinal epithelial cells

Gibson, A. R.; Diaz Ludovico, I.; Clair, G. C.; Hutchinson-Bunch, C. M.; Adkins, J. N.; Rauch, I.

2026-04-22 immunology
10.64898/2026.04.20.719683 bioRxiv
Show abstract

Inflammasomes are cytosolic innate immune sensors that, once activated by a pathogenic threat, lead to activation of the inflammatory Caspase-1. Inflammasome activation and its consequences have been studied extensively in myeloid cells and in overexpression systems. Recent studies have identified cell type specific effects that are not fully explained by the known cleavage targets of Caspase-1. Here, we identified targets of caspase cleavage using mass spectrometry in primary intestinal epithelial cells by specifically activating the NAIP-NLRC4 inflammasome. We have taken an unbiased approach and developed a novel method for analyzing mass spectrometry data for evidence of caspase activity. Our approach can also be applied to existing proteomic datasets to establish the presence of caspase activity under various biological conditions. These results lay the groundwork for future studies on mechanisms of caspase-induced processes such as intestinal epithelial cell extrusion.

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