Antisense oligonucleotides (ASOs) are a novel therapeutic strategy that targets a specific gene and suppresses its expression. The cryopyrin-associated periodic syndromes (CAPS) are a spectrum of autoinflammatory diseases characterized by systemic and tissue inflammation that is caused by heterozygous gain-of-function mutations in the nucleotide-binding and oligomerization domain-like receptor (NLR) family pyrin domain containing 3 (NLRP3) gene. The aim of this study was to investigate the efficacy of an Nlrp3-specific ASO treatment in CAPS. An Nlrp3-specific ASO was designed and tested in murine cell lines and bone marrow–derived macrophages (BMDMs) from wild-type and CAPS mouse models. Nlrp3 knock-in mice were treated in vivo with Nlrp3-specific ASO, survival was monitored, and expression of organ-specific Nlrp3 and IL-1β was measured. Nlrp3-specific ASO treatment of murine cell lines and BMDMs showed a significant downregulation of Nlrp3 and mature IL-1β protein expression. Ex vivo treatment of Nlrp3 mutant mouse-derived BMDMs with Nlrp3-specific ASO demonstrated significantly reduced IL-1β release. In vivo, Nlrp3-specific ASO treatment of Nlrp3 mutant mice prolonged survival, reduced systemic inflammation, and decreased tissue-specific expression of Nlrp3 and mature IL-1β protein. The results of this study demonstrate that Nlrp3-specific ASO treatment downregulates Nlrp3 expression and IL-1β release in CAPS models, suggesting ASO therapy as a potential treatment of CAPS and other NLRP3-mediated diseases.

The Nod-like receptor family pyrin domain–containing 3 (NLRP3) inflammasome is a cytosolic multiprotein immune sensor that is restrained from assembling at steady state. Mutations in NLRP3 associated with cryopyrin-associated periodic syndrome (CAPS) enhance inflammasome formation and activation, resulting in neutrophil-predominant systemic and tissue inflammation. Here, we investigated how gasdermin D (GSDMD) and GSDME drive interleukin-1β (IL-1β) release, pyroptosis, and autoinflammation during constitutive and canonical NLRP3 activation in neutrophils. Mass cytometry analysis of neutrophils from patients with CAPS revealed heterogeneous subtypes with differing amounts of intracellular IL-1β. In mice, neutrophil-specific expression of activating Nlrp3 mutations orthologous to those in patients with CAPS uncovered nonredundant roles for GSDMD and GSDME downstream of inflammasome activation. Combined deletion of Gsdmd and Gsdme in these CAPS models reduced IL-1β secretion, prevented pyroptosis, and mitigated autoinflammation. Loss of both GSDMD and GSDME reduced caspase-1 cleavage and pro–IL-1β processing downstream of NLRP3 activation, whereas GSDMD deficiency alone increased cleavage of GSDME and caspase-8. Clusters of GSDMD and GSDME were closely associated with one another at the plasma membrane of activated neutrophils from CAPS mice. These findings indicate that the coordinated actions of membrane-associated GSDMD and GSDME reinforce cytosolic NLRP3 inflammasome activation to promote IL-1β secretion, lytic cell death, and autoinflammation.

Metabolic dysfunction-associated steatotic liver disease (MASLD) encompasses a spectrum of liver injury ranging from simple steatosis to metabolic dysfunction-associated steatohepatitis (MASH), a progressive inflammatory state associated with hepatocyte injury, fibrosis, cirrhosis, and hepatocellular carcinoma. Increasing evidence suggests that progression to MASH is not spatially uniform across the liver. Instead, it reflects region-specific metabolic stress, inflammatory signaling, and fibrogenic remodeling along the porto-central axis of the hepatic lobule. Recent advances in spatial transcriptomics, lipidomics, proteomics, and multiplex imaging have provided new insight into how hepatocytes, immune cells, endothelial cells, and hepatic stellate cells interact within distinct hepatic microenvironments during disease progression. In this review, we discuss emerging concepts linking hepatic zonation to steatosis, inflammation, fibrosis, and extracellular matrix remodeling in MASLD and MASH. We highlight how disruption of normal lobular organization contributes to a progressive loss of metabolic compartmentalization and to the amplification of inflammatory and fibrogenic signaling. We additionally examine how spatial disease programs differ between adult and pediatric MASLD, particularly given the periportal-predominant injury patterns frequently observed in children and the current lack of pediatric-focused mechanistic studies. Finally, we discuss recently approved therapies and emerging therapeutic strategies within the context of hepatic microenvironment remodeling and disease heterogeneity. Collectively, these findings support a framework in which MASLD and MASH are spatially organized diseases driven by dynamic multicellular interactions, emphasizing the importance of incorporating zonation and tissue context into future mechanistic studies and therapeutic development.