Engineering single‐domain antibodies targeting Gasdermin E activation by the chemotherapeutic agent cis‐diaminodichloroplatinum

Abstract As mediators of pyroptosis, gasdermins (GSDMs) are closely associated with systemic cytotoxicity or so‐called side effects and are also involved in the inflammatory response during chemotherapy. Using in situ proximity ligation assay followed by sequencing ( is PLA‐seq), which we recently developed, we screened a single‐domain antibody (sdAb) library and identified several sdAbs against Gasdermin E (GSDME) that specifically recognize the N‐terminal domain (1‐270 aa) of GSDME (GSDME‐NT). One of them mitigated the release of inflammatory damage‐associated molecular patterns (DAMPs) and cytokines, including high mobility group protein b1 (Hmgb1) and interleukin‐1β (Il‐1β), in isolated mouse alveolar epithelial cells (AECs) upon chemotherapeutic agent cis ‐diaminodichloroplatinum (CDDP) treatment. Further investigation showed that this anti‐GSDME sdAb also alleviated CDDP‐induced pyroptotic cell death and lung tissue injury and decreased systemic Hmgb1 release in C57/BL6 mice, due to GSDME inactivation. Collectively, our data define an inhibitory role of the specific sdAb against GSDME, providing a potential strategy for systemically alleviating chemotherapeutic toxicities in vivo.