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Disulfiram accelerates diabetic foot ulcer healing by blocking NET formation via suppressing the NLRP3/Caspase-1/GSDMD pathway

  • Shuofei Yang
    Correspondence
    Reprint requests: Shuofei Yang, Lan Zhang, and Guanhua Xue, Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Pujian Road 160, Shanghai, 200127, China.
    Affiliations
    Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
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  • Yu Feng
    Affiliations
    Department of Endocrinology, The Second Affiliated Hospital of Soochow University, Suzhou, China
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  • Liang Chen
    Affiliations
    Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
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  • Zheyu Wang
    Affiliations
    Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
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  • Jiaquan Chen
    Affiliations
    Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
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  • Qihong Ni
    Affiliations
    Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
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  • Xiangjiang Guo
    Affiliations
    Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
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  • Lan Zhang
    Correspondence
    Reprint requests: Shuofei Yang, Lan Zhang, and Guanhua Xue, Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Pujian Road 160, Shanghai, 200127, China.
    Affiliations
    Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
    Search for articles by this author
  • Guanhua Xue
    Correspondence
    Reprint requests: Shuofei Yang, Lan Zhang, and Guanhua Xue, Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Pujian Road 160, Shanghai, 200127, China.
    Affiliations
    Department of Vascular Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
    Search for articles by this author
Published:November 03, 2022DOI:https://doi.org/10.1016/j.trsl.2022.10.008

      Abstract

      Diabetic foot ulcer (DFU) is among the most frequent complications of diabetes and is associated with significant morbidity and mortality. Excessive neutrophil extracellular traps (NETs) delay wound healing in diabetic patients. Therefore, interventions targeting NET release need to be developed to effectively prevent NET-based wound healing impairment. Gasdermin D (GSDMD), a pore-forming protein acts as a central executioner of inflammatory cell death and can activate inflammasomes in neutrophils to release NETs. A precise understanding of the mechanism underlying NET-mediated delay in diabetic wound healing may be valuable in identifying potential therapeutic targets to improve clinical outcomes. In this study, we reported that neutrophils were more susceptible to NETosis in diabetic wound environments of patients with DFU. By in vitro experiments and using in vivo mouse models of diabetic wound healing (wide-type, Nlrp3−/−, Casp-1−/−, and Gsdmd−/− mice), we demonstrated that NLRP3/caspase-1/GSDMD pathway on activation controls NET release by neutrophils in diabetic wound tissue. Furthermore, inhibition of GSDMD with disulfiram or genic deletion of Gsdmd abrogated NET formation, thereby accelerating diabetic wound healing. Disulfiram could inhibit NETs-mediated diabetic foot ulcer healing impairment by suppressing the NLRP3/Caspase-1/GSDMD pathway. In summary, our findings uncover a novel therapeutic role of disulfiram in inhibiting NET formation, which is of considerable value in accelerating wound healing in patients with DFU.

      Abbreviations:

      Diabetic foot ulcer (DFU), Neutrophil extracellular traps (NETs), Gasdermin D (GSDMD), Peptidyl arginine deiminase 4 (PAD4), Cell-free double-stranded DNA (cfDNA), neutrophil elastase (NE), Wild-type (WT), Streptozotocin (STZ), 4',6-diamidino-2-phenylindole dihydrochloride (DAPI), Phosphate buffered saline (PBS), Lipopolysaccharides (LPS)
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      References

        • Falanga V.
        Wound healing and its impairment in the diabetic foot.
        Lancet (London, England). 2005; 366: 1736-1743
        • Martin P
        • Leibovich SJ.
        Inflammatory cells during wound repair: the good, the bad and the ugly.
        Trends Cell Biol. 2005; 15: 599-607
        • Brinkmann V
        • Reichard U
        • Goosmann C
        • et al.
        Neutrophil extracellular traps kill bacteria.
        Science. 2004; 303: 1532-1535
        • Leshner M
        • Wang S
        • Lewis C
        • et al.
        PAD4 mediated histone hypercitrullination induces heterochromatin decondensation and chromatin unfolding to form neutrophil extracellular trap-like structures.
        Front Immunol. 2012; 3: 307
        • Lee KH
        • Kronbichler A
        • Park DD
        • et al.
        Neutrophil extracellular traps (NETs) in autoimmune diseases: a comprehensive review.
        Autoimmun Rev. 2017; 16: 1160-1173
        • Joshi MB
        • Ahamed R
        • Hegde M
        • Nair AS
        • Ramachandra L
        • Satyamoorthy K.
        Glucose induces metabolic reprogramming in neutrophils during type 2 diabetes to form constitutive extracellular traps and decreased responsiveness to lipopolysaccharides.
        Biochim Biophys Acta Mol Basis Dis. 2020; 1866165940
        • Hirota T
        • Levy JH
        • Iba T.
        The influence of hyperglycemia on neutrophil extracellular trap formation and endothelial glycocalyx damage in a mouse model of type 2 diabetes.
        Microcirculation. 2020; 27: e12617
        • Remijsen Q
        • Kuijpers TW
        • Wirawan E
        • Lippens S
        • Vandenabeele P
        • Vanden Berghe T.
        Dying for a cause: NETosis, mechanisms behind an antimicrobial cell death modality.
        Cell Death Differ. 2011; 18: 581-588
        • Wong SL
        • Demers M
        • Martinod K
        • et al.
        Diabetes primes neutrophils to undergo NETosis, which impairs wound healing.
        Nat Med. 2015; 21: 815-819
        • Fadini GP
        • Menegazzo L
        • Rigato M
        • et al.
        NETosis delays diabetic wound healing in mice and humans.
        Diabetes. 2016; 65: 1061-1071
        • Yang S
        • Gu Z
        • Lu C
        • et al.
        Neutrophil extracellular traps are markers of wound healing impairment in patients with diabetic foot ulcers treated in a multidisciplinary setting.
        Adv Wound Care. 2020; 9: 16-27
        • W-t He
        • H Wan
        • Hu L
        • et al.
        Gasdermin D is an executor of pyroptosis and required for interleukin-1β secretion.
        Cell Res. 2015; 25: 1285-1298
        • Matikainen S
        • Nyman TA
        • Cypryk W.
        Function and regulation of noncanonical caspase-4/5/11 inflammasome.
        J Immunol. 2020; 204: 3063-3069
        • Yu Z-W
        • Zhang J
        • Li X
        • Wang Y
        • Fu Y-H
        • Gao X-Y.
        A new research hot spot: the role of NLRP3 inflammasome activation, a key step in pyroptosis, in diabetes and diabetic complications.
        Life Sci. 2020; 240117138
        • Liu D
        • Yang P
        • Gao M
        • et al.
        NLRP3 activation induced by neutrophil extracellular traps sustains inflammatory response in the diabetic wound.
        Clin Sci (Lond). 2019; 133: 565-582
        • Yao F
        • Jin Z
        • Zheng Z
        • et al.
        HDAC11 promotes both NLRP3/caspase-1/GSDMD and caspase-3/GSDME pathways causing pyroptosis via ERG in vascular endothelial cells.
        Cell Death Discov. 2022; 8: 112
        • Cheng Q
        • Pan J
        • Zhou Z-L
        • et al.
        Caspase-11/4 and gasdermin D-mediated pyroptosis contributes to podocyte injury in mouse diabetic nephropathy.
        Acta Pharmacol Sin. 2021; 42: 954-963
        • Wang Y
        • Zhu X
        • Yuan S
        • et al.
        TLR4/NF-κB signaling induces GSDMD-related pyroptosis in tubular cells in diabetic kidney disease.
        Front Endocrinol (Lausanne). 2019; 10: 603
        • Munzer P
        • Negro R
        • Fukui S
        • et al.
        NLRP3 Inflammasome assembly in neutrophils is supported by PAD4 and promotes NETosis under sterile conditions.
        Front Immunol. 2021; 12683803
        • Sung PS
        • Peng YC
        • Yang SP
        • Chiu CH
        • Hsieh SL.
        CLEC5A is critical in Pseudomonas aeruginosa-induced NET formation and acute lung injury.
        JCI Insight. 2022; 7: e156613
        • Koppaka V
        • Thompson DC
        • Chen Y
        • et al.
        Aldehyde dehydrogenase inhibitors: a comprehensive review of the pharmacology, mechanism of action, substrate specificity, and clinical application.
        Pharmacol Rev. 2012; 64: 520-539
        • Hu JJ
        • Liu X
        • Xia S
        • et al.
        FDA-approved disulfiram inhibits pyroptosis by blocking gasdermin D pore formation.
        Nat Immunol. 2020; 21: 736-745
        • Chen KW
        • Monteleone M
        • Boucher D
        • et al.
        Noncanonical inflammasome signaling elicits gasdermin D-dependent neutrophil extracellular traps.
        Sci Immunol. 2018; 3: eaar6676
        • Silva CMS
        • Wanderley CWS
        • Veras FP
        • et al.
        Gasdermin D inhibition prevents multiple organ dysfunction during sepsis by blocking NET formation.
        Blood. 2021; 138: 2702-2713
        • Beckert S
        • Witte M
        • Wicke C
        • Konigsrainer A
        • Coerper S.
        A new wound-based severity score for diabetic foot ulcers: a prospective analysis of 1,000 patients.
        Diabetes Care. 2006; 29: 988-992
        • Hicks CW
        • Canner JK
        • Karagozlu H
        • et al.
        The Society for Vascular Surgery Wound, Ischemia, and foot Infection (WIfI) classification system correlates with cost of care for diabetic foot ulcers treated in a multidisciplinary setting.
        J Vasc Surg. 2018; 67: 1455-1462
        • Barcelos LS
        • Duplaa C
        • Krankel N
        • et al.
        Human CD133+ progenitor cells promote the healing of diabetic ischemic ulcers by paracrine stimulation of angiogenesis and activation of Wnt signaling.
        Circ Res. 2009; 104: 1095-1102
        • Caporali A
        • Meloni M
        • Miller AM
        • et al.
        Soluble ST2 is regulated by p75 neurotrophin receptor and predicts mortality in diabetic patients with critical limb ischemia.
        Arterioscler Thromb Vasc Biol. 2012; 32: e149-e160
        • Miyabe Y
        • Kim ND
        • Miyabe C
        • Luster AD.
        Studying neutrophil migration in vivo using adoptive cell transfer.
        Methods Mol Biol. 2016; 1407: 179-194
        • Shi J
        • Zhao Y
        • Wang K
        • et al.
        Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death.
        Nature. 2015; 526: 660-665
        • Sollberger G
        • Choidas A
        • Burn GL
        • et al.
        Gasdermin D plays a vital role in the generation of neutrophil extracellular traps.
        Sci Immunol. 2018; 3: eaar6689
        • Lewis HD
        • Liddle J
        • Coote JE
        • et al.
        Inhibition of PAD4 activity is sufficient to disrupt mouse and human NET formation.
        Nat Chem Biol. 2015; 11: 189-191
        • Yang S
        • Qi H
        • Kan K
        • et al.
        Neutrophil extracellular traps promote hypercoagulability in patients with sepsis.
        Shock (Augusta, Ga). 2017; 47: 132-139
        • Brem H
        • Tomic-Canic M
        Cellular and molecular basis of wound healing in diabetes.
        J Clin Invest. 2007; 117: 1219-1222
        • Aldabbous L
        • Abdul-Salam V
        • McKinnon T
        • et al.
        Neutrophil extracellular traps promote angiogenesis: evidence from vascular pathology in pulmonary hypertension.
        Arterioscler Thromb Vasc Biol. 2016; 36: 2078-2087
        • Mizuno R
        • Kawada K
        • Itatani Y
        • Ogawa R
        • Kiyasu Y
        • Sakai Y.
        The role of tumor-associated neutrophils in colorectal cancer.
        Int J Mol Sci. 2019; 20: 529
        • Zhu S
        • Yu Y
        • Ren Y
        • et al.
        The emerging roles of neutrophil extracellular traps in wound healing.
        Cell Death Dis. 2021; 12: 984
        • Das SK
        • Yuan YF
        • Li MQ.
        Specific PKC betaII inhibitor: one stone two birds in the treatment of diabetic foot ulcers.
        Biosci Rep. 2018; 38BSR20171459
        • Yang CT
        • Chen L
        • Chen WL
        • et al.
        Hydrogen sulfide primes diabetic wound to close through inhibition of NETosis.
        Mol Cell Endocrinol. 2019; 480: 74-82
        • Kaur T
        • Dumoga S
        • Koul V
        • Singh N.
        Modulating neutrophil extracellular traps for wound healing.
        Biomater Sci. 2020; 8: 3212-3223
        • Papayannopoulos V.
        Neutrophil extracellular traps in immunity and disease.
        Nat Rev Immunol. 2018; 18: 134-147
        • Adrover JM
        • Carrau L
        • Daßler-Plenker J
        • et al.
        Disulfiram inhibits neutrophil extracellular trap formation and protects rodents from acute lung injury and SARS-CoV-2 infection.
        JCI Insight. 2022; 7: e157342
        • Schmidt FI
        • Latz E.
        Jack of all trades inhibits inflammation (in sober people).
        Nat Immunol. 2020; 21: 718-719
        • Tsuchiya K.
        Inflammasome-associated cell death: Pyroptosis, apoptosis, and physiological implications.
        Microbiol Immunol. 2020; 64: 252-269
        • Ramos-Junior ES
        • Morandini AC.
        Gasdermin: a new player to the inflammasome game.
        Biomed J. 2017; 40: 313-316
        • Bitto A
        • Altavilla D
        • Pizzino G
        • et al.
        Inhibition of inflammasome activation improves the impaired pattern of healing in genetically diabetic mice.
        Br J Pharmacol. 2014; 171: 2300-2307
        • Mirza RE
        • Fang MM
        • Weinheimer-Haus EM
        • Ennis WJ
        • Koh TJ.
        Sustained inflammasome activity in macrophages impairs wound healing in type 2 diabetic humans and mice.
        Diabetes. 2014; 63: 1103-1114
        • Gan J
        • Huang M
        • Lan G
        • Liu L
        • Xu F.
        High Glucose Induces the Loss of Retinal Pericytes Partly via NLRP3-Caspase-1-GSDMD-Mediated Pyroptosis.
        Biomed Res Int. 2020; 20204510628
        • Liu P
        • Zhang Z
        • Wang J
        • Zhang X
        • Yu X
        • Li Y.
        Empagliflozin protects diabetic pancreatic tissue from damage by inhibiting the activation of the NLRP3/caspase-1/GSDMD pathway in pancreatic β cells: in vitro and in vivo studies.
        Bioengineered. 2021; 12: 9356-9366
        • Kambara H
        • Liu F
        • Zhang X
        • et al.
        Gasdermin D Exerts Anti-inflammatory Effects by Promoting Neutrophil Death.
        Cell Rep. 2018; 22: 2924-2936
        • de Buhr N
        • Bonilla MC
        • Pfeiffer J
        • et al.
        Degraded neutrophil extracellular traps promote the growth of Actinobacillus pleuropneumoniae.
        Cell Death Dis. 2019; 10: 657