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  3. Neutrophil extracellular traps formation driven by novel ELANE mutations in cyclic neutropenia patients

Neutrophil extracellular traps formation driven by novel ELANE mutations in cyclic neutropenia patients

  • Genes Immun. 2026 Jun 23. doi: 10.1038/s41435-026-00403-4.
Magdalena Rutkowska-Zapała # 1 2,  Adrianna Gałuszka-Bulaga # 1,  Jacek Plewka 3,  Marzena Lenart 1 2,  Anna Kluczewska 1 2,  Marta Surman 2,  Karolina Bukowska-Strakova 1 2,  Monika Rygielska 4,  Izabela Siemińska 1 5,  Katarzyna Baran 6,  Joanna Cichy 7,  Joanna Skrzeczyńska-Moncznik 7,  Jarosław Baran 1 2,  Maciej Siedlar 8 9,  Anna Szaflarska 10 11
Affiliations

Affiliations

  • 1 Department of Clinical Immunology, Institute of Pediatrics, Jagiellonian University Medical College, Krakow, Poland.
  • 2 Department of Clinical Immunology, Children's University Hospital, Krakow, Poland.
  • 3 Department of Crystal Chemistry and Crystal Physics, Faculty of Chemistry, Jagiellonian University, Krakow, Poland.
  • 4 Department of Pediatric Oncology and Hematology, Hematology Laboratory, University Children's Hospital, Krakow, Poland.
  • 5 Institute of Veterinary Sciences, Faculty of Veterinary Medicine, University of Agriculture in Krakow, Krakow, Poland.
  • 6 Faculty of Pharmacy, Jagiellonian University Medical College, Krakow, Poland.
  • 7 Department of Immunology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
  • 8 Department of Clinical Immunology, Institute of Pediatrics, Jagiellonian University Medical College, Krakow, Poland. [email protected].
  • 9 Department of Clinical Immunology, Children's University Hospital, Krakow, Poland. [email protected].
  • 10 Department of Clinical Immunology, Institute of Pediatrics, Jagiellonian University Medical College, Krakow, Poland. [email protected].
  • 11 Department of Clinical Immunology, Children's University Hospital, Krakow, Poland. [email protected].
  • # Contributed equally.
PMID: 42337056 DOI: 10.1038/s41435-026-00403-4
Abstract

Cyclic neutropenia (CyN) is a rare haematological disorder most often caused by mutations in the neutrophil Elastase gene (ELANE). We describe three novel ELANE mutations in CyN patients and link them to impaired neutrophil extracellular trap (NET) formation. ELANE Sequencing was performed using the Sanger method, and AlphaFold2 was used to visualize the impact of mutations on the structure of neutrophil Elastase (NE). NET formation was assessed by colocalization of MPO, NE, and DNA using fluorescence microscopy, and free DNA was quantified fluorometrically. We identified two missense substitutions (M66K, V133G) and one deletion (G192-G196del). These mutations did not significantly alter the overall NE structure but exhibited functional effects: M66K may disrupt substrate binding, V133G increases protein mobility, and the Deletion of G192-G196 may limit the opening of the catalytic pocket. NETosis was impaired in CyN patients compared with controls. Their NETs appeared less expanded, and DNA release into supernatants was reduced, reaching the lowest levels during neutropenia, even in patients with normal neutrophil counts. These abnormalities were accompanied by reduced NE activity. In sum, novel ELANE variants may contribute to altered NE function and defective NET formation in CyN, suggesting that impaired NETosis is partly independent of cyclic changes in neutrophil numbers.

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