title: NETosis Delays Diabetic Wound Healing in Mice and Humans.
abstract: Upon activation, neutrophils undergo histone citrullination by protein arginine deiminase (PAD)4, exocytosis of chromatin and enzymes as neutrophil extracellular traps (NETs), and death. In diabetes, neutrophils are primed to release NETs and die by NETosis. Although this process is a defense against infection, NETosis can damage tissue. Therefore, we examined the effect of NETosis on the healing of diabetic foot ulcers (DFUs). Using proteomics, we found that NET components were enriched in nonhealing human DFUs. In an independent validation cohort, a high concentration of neutrophil elastase in the wound was associated with infection and a subsequent worsening of the ulcer. NET components (elastase, histones, neutrophil gelatinase-associated lipocalin, and proteinase-3) were elevated in the blood of patients with DFUs. Circulating elastase and proteinase-3 were associated with infection, and serum elastase predicted delayed healing. Neutrophils isolated from the blood of DFU patients showed an increased spontaneous NETosis but an impaired inducible NETosis. In mice, skin PAD4 activity was increased by diabetes, and FACS detection of histone citrullination, together with intravital microscopy, showed that NETosis occurred in the bed of excisional wounds. PAD4 inhibition by Cl-amidine reduced NETting neutrophils and rescued wound healing in diabetic mice. Cumulatively, these data suggest that NETosis delays DFU healing.
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy Venetian Institute of Molecular Medicine, Padova, Italy gianpaolo.fadini@unipd.it.
authors.forename: Gian Paolo
authors.initials: GP
authors.lastname: Fadini
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy Venetian Institute of Molecular Medicine, Padova, Italy.
authors.forename: Lisa
authors.initials: L
authors.lastname: Menegazzo
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Rigato
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy Venetian Institute of Molecular Medicine, Padova, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Scattolini
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy Venetian Institute of Molecular Medicine, Padova, Italy.
authors.forename: Nicol
authors.initials: N
authors.lastname: Poncina
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Bruttocao
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy Venetian Institute of Molecular Medicine, Padova, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Ciciliot
authors.affiliation: Venetian Institute of Molecular Medicine, Padova, Italy Department of Physics and Astronomy, University of Padova, Padova, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Mammano
authors.affiliation: Centro Nazionale per le Ricerche Institute of Neuroscience, Padova, Italy.
authors.forename: Catalin Dacian
authors.initials: CD
authors.lastname: Ciubotaru
authors.affiliation: Foot and Ankle Clinic, Policlinico di Abano Terme, Abano Terme, Italy.
authors.forename: Enrico
authors.initials: E
authors.lastname: Brocco
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy.
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Marescotti
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Cappellari
authors.affiliation: Department of Biomedical Sciences, University of Padova, Padova, Italy Proteomic Center of Padova, University of Padova, Padova, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Arrigoni
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy Proteomic Center of Padova, University of Padova, Padova, Italy.
authors.forename: Renato
authors.initials: R
authors.lastname: Millioni
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy.
authors.forename: Saula
authors.initials: S
authors.lastname: Vigili de Kreutzenberg
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy Venetian Institute of Molecular Medicine, Padova, Italy.
authors.forename: Mattia
authors.initials: M
authors.lastname: Albiero
authors.affiliation: Department of Medicine, University of Padova, Padova, Italy Venetian Institute of Molecular Medicine, Padova, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: Avogaro
mesh_terms: D000368:Aged; D000818:Animals; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental / Q000150:complications / Q000276:immunology / Q000503:physiopathology*; D003924:Diabetes Mellitus, Type 2 / Q000150:complications / Q000276:immunology / Q000503:physiopathology*; D017719:Diabetic Foot / Q000276:immunology / Q000473:pathology / Q000503:physiopathology; D065206:Extracellular Traps / Q000502:physiology*; D005260:Female; D006801:Humans; D019272:Leukocyte Elastase / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008875:Middle Aged; D009504:Neutrophils / Q000378:metabolism; D013997:Time Factors; D014945:Wound Healing / Q000276:immunology / Q000502:physiology*
chemical_list: D019272:Leukocyte Elastase