Poster & Lightening Talk Inaugural Australian Ubiquitin Summit 2025

NEDD4L mediated Gasdermin D and E ubiquitination regulates cell death and tissue injury (129369)

Sonia S Shah 1 2 , Jantina A Manning 1 2 , Yoon Lim 1 2 , Diva Sinha 1 , Raja Ganesan 3 , Nirmal Robinson 1 2 , Emad S Alnemri 4 , Seth L Masters 5 6 , James Vince 7 , Sharad Kumar 1 2 8
  1. Centre for Cancer Biology, Adelaide, SA, Australia
  2. Adelaide University, Adelaide, SA, Australia
  3. University Hospital Cologne, TRIO Research Center, 50931 Köln, Germany
  4. Department of Biochemistry and Molecular Biology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, 19107, USA
  5. Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research, Clayton, VIC 3168, Australia
  6. The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC 3052, Australia
  7. The Department of Medical Biology, University of Melbourne, Parkville, VIC 3010, Australia
  8. Adelaide Medical School, The University of Adelaide, Adelaide, SA 5005, Australia

Gasdermins (GSDMs) are a family of pore forming proteins, that play a crucial role in various biological processes such as immune response, and have been associated with cancers, autoimmune diseases and other disorders. They are essential executors of pyroptosis, a lytic type of inflammatory cell death. Several cellular stressors such as bacterial infections, ionic imbalance, and toxins cause GSDM activation, allowing these proteins to form large membrane pores, cytokine release, and cell death. However, little is known about how these proteins are regulated and their cell specific functions. Here, we identify the ubiquitin ligase NEDD4L as a key regulator of GSDMD and GSDME, two GSDMs involved in cell death. NEDD4L interacts with and ubiquitinates both these proteins to control their stability and intracellular expression levels. Knockout of mouse Nedd4l (Nedd4-2) results in lung and kidney damage with perinatal lethality within three weeks of birth. These mice demonstrated elevated GSDMD in alveolar epithelia and increased GSDME in kidney tubular epithelia, suggesting tissue specific regulation by NEDD4L. Kidney specific Nedd4l knockout mice showed GSDMD and GSDME activation, tubular cell death and reduced kidney function after high sodium diet. NEDD4L-deficient cells were significantly more susceptible to GSDM activation and showed higher IL-1β release and increased cell death induced by NLRP3 agonists, cytotoxic agents and bacterial infection. These results demonstrate that NEDD4L regulates GSDMD and GSDME functions by preventing their accumulation and reveals an unexplored link between GSDM stability and cell death, which is distinct from current GSDM inhibitors that prevent membrane targeting.