Regulatory mechanisms of gasdermin-mediated pyroptosis

The National Institute of General Medical Sciences awarded Cincinnati Children's Hospital Medical Center $405,000 on August 1, 2026, under the Biomedical Research and Research Training program (CFDA 93.859) to conduct a five-year research program investigating the regulatory mechanisms of gasdermin-mediated pyroptosis.

The research uses functional genomics, proteomics, and live cell imaging to study how gasdermin family proteins control pyroptosis, a highly inflammatory form of programmed cell death implicated in allergic diseases, metabolic disorders, autoimmune diseases, infectious diseases, and cancer. The project builds on pilot genetic screening results on gasdermin B (GSDMB) regulation and will investigate candidate regulatory factors in pyroptosis, track the intracellular movement and trafficking of gasdermin protein fragments through live cell imaging, identify suppressors of gasdermin activity via genome-wide CRISPR screening, and construct proteomic maps of gasdermin interactomes to identify post-translational modifications critical to gasdermin biochemical activity.

Performance occurs in Cincinnati, Ohio, with the period of performance running through April 30, 2031. The award is assistance in the form of a project grant.

Abstract

The highly inflammatory programmed cell death, pyroptosis, is mediated by the gasdermin family of structurally related, pore-forming proteins (gsdma-f and pjvk) and is implicated in numerous diseases including allergic diseases, metabolic disorders, autoimmune diseases, infectious diseases, and cancer. During pyroptosis, proteolytic cleavage of cytoplasmic full-length gasdermin liberates its n-terminal fragment (gsdm-NT) from the autoinhibition by the c-terminal fragment (gsdm-CT) allowing gsdm-NT to translocate to membranes and form pores. The gasdermin membrane pores serve as channels for the extracellular release of molecules including cytokines, but sustained activation of gasdermins leads to cell lysis. However, our knowledge of the regulatory mechanisms of gasdermin-mediated pyroptosis remains incomplete. Over the next five years, we envision a research program that will address this knowledge GAP using functional genomics, proteomics, and live cell imaging. Leveraging the results of our pilot genetic screen on the regulation of gasdermin b (gsdmb)-mediated cell death, we will investigate the regulatory role of the identified candidate facilitators in pyroptosis. Moreover, through live cell imaging, we will track the movement of gsdmb-NT, gsdmb-CT, and the pyroptotic null gsdmb-NT variant inside the cells and determine how the candidate regulators influence their intracellular trafficking and function. We will also identify suppressors of gsdmb-NT activity through bidirectional genome- wide crispr activation and knockout screens. Furthermore, through high-throughput proteomics screens, we will construct a map of gsdmb-NT and gsdme-NT interactomes which can inform US of post-translational modifications critical to the gasdermins' biochemical activity. Our research program promises to advance our understanding of the regulatory mechanisms of gasdermin-mediated pyroptosis which can be exploited for the development of novel therapies targeting pyroptosis-related diseases.