Complex regulation of alarmins S100A8/A9 and secretion via gasdermin D pores exacerbates autoinflammation in familial Mediterranean fever
Selina K. Jorch, Annika McNally, Philipp Berger, Jonas Wolf, Kim Kaiser, Andrian Chetrusca, Stefanie Robeck, Isabell Pastau, Olesja Fehler, Saskia-Larissa Jauch-Speer, Sven Hermann, Michael Schäfers, Hanne Van Gorp, Apurva Kanneganti, Joke Dehoorne, Filomeen Haerynck, Federica Penco, Marco Gattorno, Jae Jin Chae, Paul Kubes, Mohamed Lamkanfi, Andy Wullaert, Markus Sperandio, Thomas Vogl, Johannes Roth, Judith Austermann
Abstract
Background Familial Mediterranean fever (FMF), caused by mutations in the pyrin-encoding MEFV gene, is characterized by uncontrolled caspase-1 activation and IL-1β secretion. A similar mechanism drives inflammation in cryopyrin-associated periodic fever syndrome (CAPS) caused by mutations in NLRP3. CAPS and FMF, however, result in largely different clinical manifestations, pointing to additional, autoinflammatory pathways involved in FMF. Another hallmark of FMF is extraordinarily high expression of S100A8 and S100A9. These alarmins are ligands of Toll-like receptor 4 and amplifiers of inflammation. However, the relevance of this inflammatory pathway for the pathogenesis of FMF is unknown. Objective This study investigated whether mutations in pyrin result in specific secretion of S100A8/A9 alarmins through gasdermin D pores' amplifying FMF pathology. Methods S100A8/A9 levels in FMF patients were quantified by enzyme-linked immunosorbent assay. In vitro models with knockout cell lines and specific protein inhibitors were used to unravel the S100A8/A9 secretion mechanism. The impact of S100A8/A9 to the pathophysiology of FMF was analyzed with FMF ( MEFV V726A/V726A ) and S100A9 −/− mouse models. Pyrin-S100A8/A9 interaction was investigated by coimmunoprecipitation, immunofluorescence, and enzyme-linked immunosorbent assay studies. Results The S100A8/A9 complexes directly interacted with pyrin. Knocking out pyrin, caspase-1, or gasdermin D inhibited the secretion of these S100 alarmins. Inflammatory S100A8/A9 dimers were inactivated by tetramer formation. Blocking this inactivation by targeted S100A9 deletion in a murine FMF model demonstrated the relevance of this novel autoinflammatory pathway in FMF. Conclusion This is the first proof that members of the S100 alarmin family are released in a pyrin/caspase-1/gasdermin D–dependent pathway and directly drive autoinflammation in vivo.