Advertisement
Journal of Investigative Dermatology Home

Antimicrobial Peptide LL-37 Drives Rosacea-Like Skin Inflammation in an NLRP3-Dependent Manner

  • Sung-Hyun Yoon
    Affiliations
    Department of Microbiology and Immunology, Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea
    Search for articles by this author
  • Inhwa Hwang
    Affiliations
    Department of Microbiology and Immunology, Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea
    Search for articles by this author
  • Eunju Lee
    Affiliations
    Department of Microbiology and Immunology, Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea
    Search for articles by this author
  • Hyo-Joung Cho
    Affiliations
    Department of Microbiology and Immunology, Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea
    Search for articles by this author
  • Ju Hee Ryu
    Affiliations
    Theragnosis Research Center, Biomedical Research Division, Korea Institute of Science and Technology (KIST), Seoul, Korea
    Search for articles by this author
  • Tae-Gyun Kim
    Affiliations
    Department of Microbiology and Immunology, Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea

    Department of Dermatology, Yonsei University College of Medicine, Seoul, Korea
    Search for articles by this author
  • Je-Wook Yu
    Correspondence
    Correspondence: Je-Wook Yu, Department of Microbiology and Immunology, Yonsei University College of Medicine, 50-1 Yonsei-ro, Seodaemun-gu, Seoul 03722, Korea.
    Affiliations
    Department of Microbiology and Immunology, Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea
    Search for articles by this author
Published:March 17, 2021DOI:https://doi.org/10.1016/j.jid.2021.02.745
      Rosacea is a chronic inflammatory skin disease characterized by immune response–dependent erythema and pustules. Although the precise etiology of rosacea remains elusive, its pathogenesis is reportedly associated with an increased level of antimicrobial peptide LL-37. However, molecular mechanisms underlying the progression of rosacea via LL-37 remain poorly understood. Here, we examined the potential role of LL-37 in rosacea-like skin inflammatory phenotypes at a molecular level. Our in vitro data demonstrated that LL-37 promotes NLRP3-mediated inflammasome activation in lipopolysaccharide-primed macrophages, indicated by the processing of caspase-1 and IL-1β. LL-37 was internalized into the cytoplasm of macrophages through P2X7 receptor–mediated endocytosis. Intracellular LL-37 triggered the assembly and activation of NLRP3-ASC inflammasome complex by facilitating lysosomal destabilization. Consistent with these in vitro results, intradermal LL-37 administration induced in vivo caspase-1 activation and ASC speck formation in the skin of Nlrp3-expressing, but not in Nlrp3-deficient, mice. Intradermal injection of LL-37 elicited profound recruitment of inflammatory Gr1+ cells and subsequent skin inflammation. However, LL-37–induced rosacea-like skin inflammation was significantly abrogated in Nlrp3-deficient mice. Furthermore, an NLRP3-specific inhibitor, MCC950, markedly reduced LL-37–triggered rosacea-like phenotypes. Taken together, our findings clearly indicate that NLRP3 inflammasome activation plays a crucial role in LL-37–induced skin inflammation and rosacea pathogenesis.

      Graphical abstract

      Abbreviations:

      ATP (adenosine triphosphate), BMDM (bone marrow–derived macrophage), hCAP18 (human CAP18), K+ (potassium ion), LPS (lipopolysaccharide), TLR (toll-like receptor)
      To read this article in full you will need to make a payment
      Purchase one-time access
      Society Members (SID/ESDR), remember to log in for access.
      Subscribe to Journal of Investigative Dermatology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Alexis A.F.
        • Callender V.D.
        • Baldwin H.E.
        • Desai S.R.
        • Rendon M.I.
        • Taylor S.C.
        Global epidemiology and clinical spectrum of rosacea, highlighting skin of color: review and clinical practice experience.
        J Am Acad Dermatol. 2019; 80: 1722-1729.e7
        • Di Nardo A.
        • Braff M.H.
        • Taylor K.R.
        • Na C.
        • Granstein R.D.
        • McInturff J.E.
        • et al.
        Cathelicidin antimicrobial peptides block dendritic cell TLR4 activation and allergic contact sensitization.
        J Immunol. 2007; 178: 1829-1834
        • Dinarello C.A.
        Blocking IL-1 in systemic inflammation.
        J Exp Med. 2005; 201: 1355-1359
        • Dombrowski Y.
        • Peric M.
        • Koglin S.
        • Kammerbauer C.
        • Göss C.
        • Anz D.
        • et al.
        Cytosolic DNA triggers inflammasome activation in keratinocytes in psoriatic lesions.
        Sci Transl Med. 2011; 3: 82ra38
        • Dombrowski Y.
        • Schauber J.
        Cathelicidin LL-37: a defense molecule with a potential role in psoriasis pathogenesis.
        Exp Dermatol. 2012; 21: 327-330
        • Dürr U.H.
        • Sudheendra U.S.
        • Ramamoorthy A.
        LL-37, the only human member of the cathelicidin family of antimicrobial peptides.
        Biochim Biophys Acta. 2006; 1758: 1408-1425
        • Elssner A.
        • Duncan M.
        • Gavrilin M.
        • Wewers M.D.
        A novel P2X7 receptor activator, the human cathelicidin-derived peptide LL37, induces IL-1 beta processing and release.
        J Immunol. 2004; 172: 4987-4994
        • Fernandes-Alnemri T.
        • Wu J.
        • Yu J.W.
        • Datta P.
        • Miller B.
        • Jankowski W.
        • et al.
        The pyroptosome: a supramolecular assembly of ASC dimers mediating inflammatory cell death via caspase-1 activation.
        Cell Death Differ. 2007; 14: 1590-1604
        • Fernandes-Alnemri T.
        • Yu J.W.
        • Datta P.
        • Wu J.
        • Alnemri E.S.
        AIM2 activates the inflammasome and cell death in response to cytoplasmic DNA.
        Nature. 2009; 458: 509-513
        • Fernandes-Alnemri T.
        • Yu J.W.
        • Juliana C.
        • Solorzano L.
        • Kang S.
        • Wu J.
        • et al.
        The AIM2 inflammasome is critical for innate immunity to Francisella tularensis.
        Nat Immunol. 2010; 11: 385-393
        • Fukumoto K.
        • Nagaoka I.
        • Yamataka A.
        • Kobayashi H.
        • Yanai T.
        • Kato Y.
        • et al.
        Effect of antibacterial cathelicidin peptide CAP18/LL-37 on sepsis in neonatal rats.
        Pediatr Surg Int. 2005; 21: 20-24
        • Gläser R.
        • Navid F.
        • Schuller W.
        • Jantschitsch C.
        • Harder J.
        • Schröder J.M.
        • et al.
        UV-B radiation induces the expression of antimicrobial peptides in human keratinocytes in vitro and in vivo.
        J Allergy Clin Immunol. 2009; 123: 1117-1123
        • He Y.
        • Hara H.
        • Núñez G.
        Mechanism and regulation of NLRP3 inflammasome activation.
        Trends Biochem Sci. 2016; 41: 1012-1021
        • Heneka M.T.
        • Kummer M.P.
        • Stutz A.
        • Delekate A.
        • Schwartz S.
        • Vieira-Saecker A.
        • et al.
        NLRP3 is activated in Alzheimer’s disease and contributes to pathology in APP/PS1 mice.
        Nature. 2013; 493: 674-678
        • Hu Z.
        • Murakami T.
        • Suzuki K.
        • Tamura H.
        • Kuwahara-Arai K.
        • Iba T.
        • et al.
        Antimicrobial cathelicidin peptide LL-37 inhibits the LPS/ATP-induced pyroptosis of macrophages by dual mechanism.
        PLoS One. 2014; 9: e85765
        • Kahlenberg J.M.
        • Carmona-Rivera C.
        • Smith C.K.
        • Kaplan M.J.
        Neutrophil extracellular trap-associated protein activation of the NLRP3 inflammasome is enhanced in lupus macrophages.
        J Immunol. 2013; 190: 1217-1226
        • Kim J.Y.
        • Kim Y.J.
        • Lim B.J.
        • Sohn H.J.
        • Shin D.
        • Oh S.H.
        Increased expression of cathelicidin by direct activation of protease-activated receptor 2: possible implications on the pathogenesis of rosacea.
        Yonsei Med J. 2014; 55: 1648-1655
        • Kim M.
        • Kim K.E.
        • Jung H.Y.
        • Jo H.
        • Jeong S.W.
        • Lee J.
        • et al.
        Recombinant erythroid differentiation regulator 1 inhibits both inflammation and angiogenesis in a mouse model of rosacea.
        Exp Dermatol. 2015; 24: 680-685
        • Ko Y.J.
        • Lee J.W.
        • Yang E.J.
        • Jang N.
        • Park J.
        • Jeon Y.K.
        • et al.
        Non-invasive in vivo imaging of caspase-1 activation enables rapid and spatiotemporal detection of acute and chronic inflammatory disorders.
        Biomaterials. 2020; 226: 119543
        • Lee E.
        • Hwang I.
        • Park S.
        • Hong S.
        • Hwang B.
        • Cho Y.
        • et al.
        MPTP-driven NLRP3 inflammasome activation in microglia plays a central role in dopaminergic neurodegeneration.
        Cell Death Differ. 2019; 26: 213-228
        • McHugh B.J.
        • Wang R.
        • Li H.N.
        • Beaumont P.E.
        • Kells R.
        • Stevens H.
        • et al.
        Cathelicidin is a "fire alarm", generating protective NLRP3-dependent airway epithelial cell inflammatory responses during infection with Pseudomonas aeruginosa.
        PLoS Pathog. 2019; 15e1007694
        • Mertens M.
        • Singh J.A.
        Anakinra for rheumatoid arthritis: a systematic review.
        J Rheumatol. 2009; 36: 1118-1125
        • Mookherjee N.
        • Brown K.L.
        • Bowdish D.M.
        • Doria S.
        • Falsafi R.
        • Hokamp K.
        • et al.
        Modulation of the TLR-mediated inflammatory response by the endogenous human host defense peptide LL-37.
        J Immunol. 2006; 176: 2455-2464
        • Moreno-Angarita A.
        • Aragón C.C.
        • Tobón G.J.
        Cathelicidin LL-37: A new important molecule in the pathophysiology of systemic lupus erythematosus.
        J Transl Autoimmun. 2020; 3: 100029
        • Morizane S.
        • Gallo R.L.
        Antimicrobial peptides in the pathogenesis of psoriasis.
        J Dermatol. 2012; 39: 225-230
        • Muñoz-Planillo R.
        • Kuffa P.
        • Martínez-Colón G.
        • Smith B.L.
        • Rajendiran T.M.
        • Núñez G.
        K+ efflux is the common trigger of NLRP3 inflammasome activation by bacterial toxins and particulate matter.
        Immunity. 2013; 38: 1142-1153
        • Niyonsaba F.
        • Kiatsurayanon C.
        • Chieosilapatham P.
        • Ogawa H.
        Friends or foes? Host defense (antimicrobial) peptides and proteins in human skin diseases.
        Exp Dermatol. 2017; 26: 989-998
        • Petrasek J.
        • Bala S.
        • Csak T.
        • Lippai D.
        • Kodys K.
        • Menashy V.
        • et al.
        IL-1 receptor antagonist ameliorates inflammasome-dependent alcoholic steatohepatitis in mice.
        J Clin Invest. 2012; 122: 3476-3489
        • Salzer S.
        • Kresse S.
        • Hirai Y.
        • Koglin S.
        • Reinholz M.
        • Ruzicka T.
        • et al.
        Cathelicidin peptide LL-37 increases UVB-triggered inflammasome activation: possible implications for rosacea.
        J Dermatol Sci. 2014; 76: 173-179
        • Schaller M.
        • Almeida L.M.
        • Bewley A.
        • Cribier B.
        • Dlova N.C.
        • Kautz G.
        • et al.
        Rosacea treatment update: recommendations from the global Rosacea COnsensus (ROSCO) panel.
        Br J Dermatol. 2017; 176: 465-471
        • Sobolewska B.
        • Angermair E.
        • Deuter C.
        • Doycheva D.
        • Kuemmerle-Deschner J.
        • Zierhut M.
        NLRP3 A439V mutation in a large family with cryopyrin-associated periodic syndrome: description of ophthalmologic symptoms in correlation with other organ symptoms.
        J Rheumatol. 2016; 43: 1101-1106
        • Sørensen O.E.
        • Follin P.
        • Johnsen A.H.
        • Calafat J.
        • Tjabringa G.S.
        • Hiemstra P.S.
        • et al.
        Human cathelicidin, hCAP-18, is processed to the antimicrobial peptide LL-37 by extracellular cleavage with proteinase 3.
        Blood. 2001; 97: 3951-3959
        • Tang X.
        • Basavarajappa D.
        • Haeggström J.Z.
        • Wan M.
        P2X7 receptor regulates internalization of antimicrobial peptide LL-37 by human macrophages that promotes intracellular pathogen clearance.
        J Immunol. 2015; 195: 1191-1201
        • van Zuuren E.J.
        • Fedorowicz Z.
        • Carter B.
        • van der Linden M.M.
        • Charland L.
        Interventions for rosacea.
        Cochrane Database Syst Rev. 2015; : CD003262
        • Vandanmagsar B.
        • Youm Y.H.
        • Ravussin A.
        • Galgani J.E.
        • Stadler K.
        • Mynatt R.L.
        • et al.
        The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance.
        Nat Med. 2011; 17: 179-188
        • Yamasaki K.
        • Di Nardo A.
        • Bardan A.
        • Murakami M.
        • Ohtake T.
        • Coda A.
        • et al.
        Increased serine protease activity and cathelicidin promotes skin inflammation in rosacea.
        Nat Med. 2007; 13: 975-980
        • Yamasaki K.
        • Gallo R.L.
        The molecular pathology of rosacea.
        J Dermatol Sci. 2009; 55: 77-81
        • Yamasaki K.
        • Gallo R.L.
        Rosacea as a disease of cathelicidins and skin innate immunity.
        J Investig Dermatol Symp Proc. 2011; 15: 12-15
        • Yang D.
        • Chen Q.
        • Schmidt A.P.
        • Anderson G.M.
        • Wang J.M.
        • Wooters J.
        • et al.
        LL-37, the neutrophil granule- and epithelial cell-derived cathelicidin, utilizes formyl peptide receptor-like 1 (FPRL1) as a receptor to chemoattract human peripheral blood neutrophils, monocytes, and T cells.
        J Exp Med. 2000; 192: 1069-1074
        • Yu J.W.
        • Lee M.S.
        Mitochondria and the NLRP3 inflammasome: physiological and pathological relevance.
        Arch Pharm Res. 2016; 39: 1503-1518

      Supplementary Reference

        • Fernandes-Alnemri T.
        • Yu J.W.
        • Datta P.
        • Wu J.
        • Alnemri E.S.
        AIM2 activates the inflammasome and cell death in response to cytoplasmic DNA.
        Nature. 2009; 458: 509-513