Activity of granulomatosis with polyangiitis and its correlation with mTOR phosphoproteomics in neutrophils

Front Immunol. 2023 Aug 31:14:1227369. doi: 10.3389/fimmu.2023.1227369. eCollection 2023.

Abstract

Introduction: Granulomatosis with polyangiitis (GPA) is a small vessel vasculitis with a complex pathomechanism. Organ damage in GPA is also mediated by extracellular trap formation (NETosis). We analyzed the functional status of phosphoproteins modulating NETosis in neutrophils by the mammalian target of rapamycin (mTOR) pathway in GPA along with NETosis biomarkers.

Methods: Phosphoproteins levels measured in isolated neutrophils from 42 patients with GPA (exacerbation n=21; remission n=21) and 21 healthy controls were compared to serum biomarkers of the disease.

Results: Neutrophils in active disease manifested lowered levels of phosphorylated mTOR(Ser2448), PTEN(Ser380) and ULK1(Ser555), whereas phosphorylated GSK-3α/β(Ser21/Ser9) was elevated. Exacerbation of GPA was characterized by elevated neutrophil dsDNA in serum, circulating mitochondrial DNA, and DNA-MPO complexes. A significant negative correlation between mTOR or PTEN phosphoproteins and biomarkers of GPA activity was also present, reflecting the clinical activity score of GPA. Positive correlations between phosphorylated GSK-3 α/β and circulating mtDNA, DNA-MPO complexes, neutrophil-released dsDNA, or circulating proteins were also significant. Increased serum levels of IGFBP-2, TFF-3, CD147, and CHI3L1 accompanied GPA exacerbation, whereas DPP-IV levels were the lowest in active GPA. Using a principal component analysis basigin, PTEN and mTOR had the highest loadings on the discrimination function, allowing classification between active, remission, and control subjects with 98% performance.

Conclusions: We present evidence that inhibited mTOR signaling accompanies NETosis in patients with GPA. The functional status of phosphoproteins suggests simultaneous activation of NETosis and autophagy. These results give rise to the study of autophagy as a mechanism underlying granuloma formation in GPA.

Keywords: autophagy; granulomatosis with polyangiitis; mammalian target of rapamycin; mitochondrial DNA; neutrophil extracellular traps; neutrophils.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Mitochondrial
  • Glycogen Synthase Kinase 3
  • Granulomatosis with Polyangiitis*
  • Humans
  • Leukocyte Disorders*
  • Neutrophils
  • Signal Transduction
  • TOR Serine-Threonine Kinases

Substances

  • Glycogen Synthase Kinase 3
  • TOR Serine-Threonine Kinases
  • DNA, Mitochondrial

Grants and funding

Study supported by the National Centre of Science in Poland- grant number: NCN 2018/31/B/NZ6/03898. Part of the analysis was carried out with the use of research infrastructure financed by the Polish Operating Programme for Intelligent Development POIR 4.2 project no. POIR.04.02.00–00-D023/20.