Gasdermin D mediates a fast transient release of ATP after NLRP3 inflammasome activation before ninjurin 1–induced lytic cell death

Summary Pyroptosis is a lytic cell death triggered by the cleavage of gasdermin (GSDM) proteins and subsequent pore formation by the N-terminal domain oligomerization in the plasma membrane. GSDMD is cleaved by caspase-1/-4/-5/-11 upon inflammasome activation and mediates IL-1β and IL-18 release. GSDMD pores favors ninjurin-1 (NINJ1) induced plasma membrane rupture and cell death. Here, we demonstrate that GSDMD mediates early ATP release upon NLRP3 inflammasome activation independently of NINJ1, occurring before IL-1β release and cell death and constituting an early danger signal. The release of ATP is a transient signal terminated before the cells continue to permeabilize and die. The different N-terminal of GSDMA to E are also able to release ATP and induce monocyte migration towards pyroptotic cells. This study reveals ATP release as an early, and transient danger signal depending on GSDMD plasma membrane permeabilization, independently of the late stages of lytic cell death. Competing Interest Statement PP declares that he is an inventor in a patent filled on March 2020 by the Fundacion para la Formacion e Investigacion Sanitaria de la Region de Murcia (PCT/EP2020/056729) for a method to identify patients with sepsis and NLRP3-disfunction, being consultant of Viva in vitro diagnostics SL. PP, LH-N and DA-B are co-founders of Viva in vitro diagnostics SL, but declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The remaining authors declare no competing interests. Footnotes ↵* Sharing first authorship.