NET Proteomic Profiling Reveals New Pathways Potentially Implicated in Dendritic Cell-Mediated Inflammation in DADA2...

Area of Science:

• Immunology
• Autoinflammatory Diseases
• Proteomics

Background:

• Deficiency of Adenosine Deaminase 2 (DADA2) is an autoinflammatory condition linked to vasculopathy and immunodeficiency.
• Neutrophil extracellular traps (NETs) are increasingly recognized for their role in DADA2 pathogenesis.
• Understanding NETs' immunogenic potential is crucial for DADA2 research.

Purpose of the Study:

• To characterize the proteomic profile of NETs in DADA2 patients compared to healthy donors (HD) and Polyarteritis Nodosa (PAN) patients.
• To investigate the functional impact of DADA2 NETs on dendritic cells (DCs).

Main Methods:

• Analysis of NETosis using Imaging Flow Cytometry in 23 DADA2 patients.
• Quantification of NETs remnants and DNAse activity in plasma via ELISA and DNA digestion assays.
• Quantitative proteomics and network analysis of NETs from DADA2, PAN, and HD groups.
• Flow cytometry analysis of circulating and monocyte-derived DCs.

Main Results:

• DADA2 patients exhibited significantly increased suicidal NETosis.
• Abnormal levels and activity of DNAse enzymes were observed in DADA2 patients.
• Proteomic analysis revealed 1356 identified proteins, with over a hundred differentially modulated in DADA2 NETs compared to controls.
• DADA2 NETs demonstrated a heightened capacity to stimulate monocyte-derived DCs in vitro.

Conclusions:

• Distinct protein profiles were identified in DADA2 NETs compared to PAN/HD NETs.
• These unique protein signatures in DADA2 NETs may contribute to activating inflammatory pathways in DCs.
• This finding suggests a potential mechanism for DADA2 pathogenesis involving NET-mediated immune cell activation.