Aberrant ceramide accumulation fuels eosinophilic inflammation by promoting eosinophil extracellular trap formation and release in chronic rhinosinusitis with nasal polyps - 11/12/25

Doi : 10.1016/j.jaci.2025.11.004

Pengda Fang, MD ^a,^b,^c, Mengzhen Wu, MD ^a,^b,^c, Junhai Chen, MD ^a,^b,^c, Wenlong Li, MD ^a,^b,^c, Yurong Bai, MD, PhD ^a, Wenyi Chen, BS ^a,^b,^c, Yue Li, MD, PhD ^a, Xinyue Wang, MD, PhD ^d, Zhenhao Xiao, MD ^a,^b,^c, Tong Wu, BS ^b,^c, Qintai Yang, MD, PhD ^a,^b,^c,^⁎ , Yana Zhang, MD, PhD ^a,^b,^c,^⁎
^a Department of Otolaryngology-Head and Neck Surgery, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China
^b Department of Allergy, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China
^c Key Laboratory of Airway Inflammatory Disease Research and Innovative Technology Translation, Guangzhou, China
^d Department of Otolaryngology, Head and Neck Surgery, The First Affiliated Hospital of Medical College of Zhejiang University, Hangzhou, China

^∗Corresponding authors: Yana Zhang, MD, PhD, or Qintai Yang, MD, PhD, Department of Otolaryngology-Head and Neck Surgery, The Third Affiliated Hospital of Sun Yat-Sen University, No. 600 Tianhe Rd, Guangzhou, Guangdong, 510630, China.Department of Otolaryngology-Head and Neck SurgeryThe Third Affiliated Hospital of Sun Yat-Sen UniversityNo. 600 Tianhe RdGuangzhouGuangdong510630China

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Graphical abstract

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Abstract

Background

Eosinophilic inflammation represents a hallmark pathologic feature of chronic rhinosinusitis with nasal polyps (CRSwNP). Although ceramides, the central sphingolipid metabolites, are implicated in asthma pathogenesis, their mechanistic contributions to CRSwNP remain poorly elucidated.

Objective

We sought to investigate the functions and associated mechanisms of ceramide in the pathogenesis of CRSwNP.

Methods

Immunofluorescence and absolute quantitative lipidomics analysis were performed to evaluate the levels of ceramide and its species in nasal biopsy samples. In vitro culture and stimulation of purified eosinophils from human peripheral blood to validate the function of ceramides. Small interfering RNA transfection and inhibition assays to determine the molecular mechanism.

Results

The expression levels of ceramide and its binding protein were enriched in eosinophilic CRSwNP. Increased Cer(18:1_24:1) was related with eosinophil extracellular trap (EET) counts and disease severity in patients with eosinophilic CRSwNP. Cer(18:1_24:1) potently induced eosinophils activation by promoting EET formation and release. Mechanistically, Cer(18:1_24:1)-induced elevated levels of reactive oxygen species (ROS)/mitochondrial ROS (mROS) promoted EET formation by upregulating PAD4/CitH3 on one hand, and drove the oligomerization of the N terminal of gasdermin D (GSDMD-N) and membrane pore formation to facilitate the EET release on the other hand. Furthermore, ceramide-binding protein PP2AC inhibition blocked EET formation and release by reduction of ROS/mROS production. Interestingly, selenium supplementation mitigated Cer(18:1_24:1)-induced EET production and release by decreasing production of ROS/mROS.

Conclusions

Aberrant ceramide accumulation fuels eosinophilic inflammation by promoting EET formation and release through the PP2AC-ROS/mROS-GSDMD-N pathway, which may contribute to the severity and progression of eosinophilic CRSwNP. These mechanistic insights may provide novel therapeutic strategies for eosinophil-driven inflammatory disorders.

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Key words : Ceramide, nasal polyps, eosinophil extracellular traps, mitochondrial ROS, membrane pores

Abbreviations used : CerS, CitH3, CRS, CRSwNP, DPI, ECP, EET, ENP, EOS, GSDMD, GSDMD-N, Met, MFI, mROS, MT, mtDNA, NAC, Nec, nENP, NP, 8-OHdG, Ox-mtDNA, PAD4, PMA, PP, PP2AC, ROS, Se-Met, siRNA, SL, SPT