ZBTB16 controls the onset of Clostridium difficile colitis through the Pyrin inflammasome

Clostridium difficile infection (CDI) is a leading cause of antibiotic-associated diarrhea and pseudomembranous colitis, and there remains a significant unmet need for therapies specifically targeting C. difficile. The Pyrin inflammasome, activated by bacterial toxins, plays a critical role in driving macrophage-mediated intestinal inflammation during CDI. In this study, we report that myeloid-specific deficiency of Zbtb16 protects mice from C. difficile-induced colitis by attenuating IL-1β-dependent inflammatory signaling. Mechanistic studies revealed that Zbtb16 deletion disrupts ASC oligomerization and speck formation, thereby selectively inhibiting inflammasome assembly and reducing mature IL-1β production in macrophages stimulated with C. difficile culture supernatant or purified TcdB toxin. Importantly, pharmacological degradation of ZBTB16 using the cereblon E3 ligase modulating drug CC-3060 significantly ameliorated colitis severity in a murine model of CDI. Our findings establish ZBTB16 as a key regulator of Pyrin inflammasome activation in macrophages, highlighting the therapeutic promise of ZBTB16 degradation as a novel strategy for treating CDI.