Ingenol mebutate induces a tumor cell-directed inflammatory response and antimicrobial peptides thereby promoting rapid tumor destruction and wound healing
Tóm tắt
Ingenol mebutat (IM)-gel is effective for the topical treatment of epithelial tumors, including actinic keratoses (AKs) or anogenital warts (AGW). AK patients treated with IM develop intensified inflammatory reactions on sights of prior clinical visible or palpable AKs as compared to the surrounding actinically damaged skin, suggesting the induction of a tumor cell-directed inflammation. AGW patients treated with IM develop even stronger inflammatory reactions with large erosions, suggesting a directed inflammatory response against HPV-infected keratinocytes. Of note, even widespread erosions heal very fast without any superinfections. Here, we set out to elucidate underlying molecular and cellular mechanisms of these clinical observations. The effects of IM (10−9–10−5 M) on the expression and translation of a comprehensive set of chemokines (CXCL1, CXCL8, CXCL9, CXCL10, CXCL11, CXCL14, CCL2, CCL5, CCL20, CCL27) and antimicrobial peptides (AMP) (HBD1, HBD2, HBD3, LL37, RNase7) were analyzed in primary human epithelial keratinocytes (HEK) and a set of epithelial cancer cell lines by RT-qPCR and ELISA in vitro. To study the possible effects of different concentrations of IM on migratory, respectively wound healing responses, an in vitro scratch assay was conducted on HEK. Ingenol mebutat significantly and dose-dependently induced the expression of proinflammatory chemokines (CXCL8, CCL2) and AMP (RNase7, HBD3) in HEK and epithelial cancer cell lines. A significantly stronger induction of CXCL8 and CCL2 was observed in our tested tumor cells as compared to HEK. We did not observe any significant effect of IM on HEK migration, respectively wound healing responses in vitro for any tested concentration (10−9, 10−8, 10−6 M) except 10−7 M, which induced a significant inhibition. Our data suggest that tumor cells are more susceptible to IM as compared to differentiated HEK. This is evident by a stronger IM-mediated induction of proinflammatory chemokines in tumor cells, which may result in a tumor cell-directed inflammatory response and rapid tumor destruction. In addition, IM induces AMP in keratinocytes and seems not to severely interfere with keratinocyte migration, which contributes to a fast and uncomplicated wound healing. Surprising is a selective inhibition of keratinocyte migration by IM at the concentration of 10−7 M pointing to very dose depending biological effects, induced by IM.
Tài liệu tham khảo
citation_journal_title=N Engl J Med; citation_title=Ingenol mebutate gel for actinic keratosis; citation_author=M Lebwohl, N Swanson, LL Anderson, A Melgaard, Z Xu, B Berman; citation_volume=366; citation_publication_date=2012; citation_pages=1010-1019; citation_doi=10.1056/NEJMoa1111170; citation_id=CR1
citation_journal_title=J Am Acad Dermatol; citation_title=Dual mechanism of action of ingenol mebutate gel for topical treatment of actinic keratoses: rapid lesion necrosis followed by lesion-specific immune response; citation_author=RH Rosen, AK Gupta, SK Tyring; citation_volume=66; citation_publication_date=2012; citation_pages=486-493; citation_doi=10.1016/j.jaad.2010.12.038; citation_id=CR2
citation_journal_title=Mol Cancer Ther; citation_title=Ingenol 3-angelate induces dual modes of cell death and differentially regulates tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in melanoma cells; citation_author=SK Gillespie, XD Zhang, P Hersey; citation_volume=3; citation_publication_date=2004; citation_pages=1651-1658; citation_id=CR3
citation_journal_title=Cancer Res; citation_title=Antitumor activity of 3-ingenyl angelate: plasma membrane and mitochondrial disruption and necrotic cell death; citation_author=SM Ogbourne, A Suhrbier, B Jones, SJ Cozzi, GM Boyle, M Morris, D McAlpine, J Johns, TM Scott, KP Sutherland; citation_volume=64; citation_publication_date=2004; citation_pages=2833-2839; citation_doi=10.1158/0008-5472.CAN-03-2837; citation_id=CR4
citation_journal_title=J Drugs Dermatol; citation_title=Ingenol mebutate: induced cell death patterns in normal and cancer epithelial cells; citation_author=M Stahlhut, M Bertelsen, M Hoyer-Hansen, N Svendsen, AH Eriksson, JM Lord, D Scheel-Toellner, SP Young, JR Zibert; citation_volume=11; citation_publication_date=2012; citation_pages=1181-1192; citation_id=CR5
citation_journal_title=J Immunol; citation_title=Neutrophils are a key component of the antitumor efficacy of topical chemotherapy with ingenol-3-angelate; citation_author=JM Challacombe, A Suhrbier, PG Parsons, B Jones, P Hampson, D Kavanagh, GE Rainger, M Morris, JM Lord, TT Le; citation_volume=177; citation_publication_date=2006; citation_pages=8123-8132; citation_doi=10.4049/jimmunol.177.11.8123; citation_id=CR6
citation_journal_title=PLoS ONE; citation_title=Tumor-preferential induction of immune responses and epidermal cell death in actinic keratoses by ingenol mebutate; citation_author=S Emmert, HA Haenssle, JR Zibert, M Schon, A Hald, MH Hansen, T Litman, MP Schon; citation_volume=11; citation_publication_date=2016; citation_pages=e0160096; citation_doi=10.1371/journal.pone.0160096; citation_id=CR7
citation_journal_title=Blood; citation_title=PEP005, a selective small-molecule activator of protein kinase C, has potent antileukemic activity mediated via the delta isoform of PKC; citation_author=P Hampson, H Chahal, F Khanim, R Hayden, A Mulder, LK Assi, CM Bunce, JM Lord; citation_volume=106; citation_publication_date=2005; citation_pages=1362-1368; citation_doi=10.1182/blood-2004-10-4117; citation_id=CR8
citation_journal_title=Cancer Immunol Immunother; citation_title=The anti-tumor agent, ingenol-3-angelate (PEP005), promotes the recruitment of cytotoxic neutrophils by activation of vascular endothelial cells in a PKC-delta dependent manner; citation_author=P Hampson, D Kavanagh, E Smith, K Wang, JM Lord, G Ed Rainger; citation_volume=57; citation_publication_date=2008; citation_pages=1241-1251; citation_doi=10.1007/s00262-008-0458-9; citation_id=CR9
citation_journal_title=Cancer Res; citation_title=Characterization of the interaction of ingenol 3-angelate with protein kinase C; citation_author=N Kedei, DJ Lundberg, A Toth, P Welburn, SH Garfield, PM Blumberg; citation_volume=64; citation_publication_date=2004; citation_pages=3243-3255; citation_doi=10.1158/0008-5472.CAN-03-3403; citation_id=CR10
citation_journal_title=Cancer Res; citation_title=The skin cancer chemotherapeutic agent ingenol-3-angelate (PEP005) is a substrate for the epidermal multidrug transporter (ABCB1) and targets tumor vasculature; citation_author=L Li, S Shukla, A Lee, SH Garfield, DJ Maloney, SV Ambudkar, SH Yuspa; citation_volume=70; citation_publication_date=2010; citation_pages=4509-4519; citation_doi=10.1158/0008-5472.CAN-09-4303; citation_id=CR11
citation_journal_title=J Dtsch Dermatol Ges; citation_title=Successful treatment of perianal condylomata acuminata with ingenol mebutate gel; citation_author=SA Braun, PA Gerber; citation_volume=14; citation_publication_date=2016; citation_pages=616-617; citation_id=CR12
citation_journal_title=J Am Acad Dermatol; citation_title=Ingenol mebutate for the management of genital warts in sensitive anatomic locations; citation_author=SA Braun, PA Gerber; citation_volume=77; citation_publication_date=2017; citation_pages=e9-e10; citation_doi=10.1016/j.jaad.2017.01.034; citation_id=CR13
citation_journal_title=Hautarzt; citation_title=Successful therapy of condylomata acuminata with ingenol mebutate; citation_author=SA Braun, TM Jansen, B Homey, PA Gerber; citation_volume=66; citation_publication_date=2015; citation_pages=223-225; citation_doi=10.1007/s00105-015-3593-1; citation_id=CR14
citation_journal_title=J Eur Acad Dermatol Venereol; citation_title=Ingenol mebutate gel is effective against anogenital warts—a case series in 17 patients; citation_author=RE Schopf; citation_volume=30; citation_publication_date=2015; citation_pages=1041-1043; citation_doi=10.1111/jdv.13097; citation_id=CR15
citation_journal_title=PLoS ONE; citation_title=Systematic identification and characterization of novel human skin-associated genes encoding membrane and secreted proteins; citation_author=PA Gerber, P Hevezi, BA Buhren, C Martinez, H Schrumpf, M Gasis, S Grether-Beck, J Krutmann, B Homey, A Zlotnik; citation_volume=8; citation_publication_date=2013; citation_pages=e63949; citation_doi=10.1371/journal.pone.0063949; citation_id=CR16
citation_journal_title=Mol Cancer; citation_title=Inhibition of oesophageal squamous cell carcinoma progression by in vivo targeting of hyaluronan synthesis; citation_author=S Twarock, T Freudenberger, E Poscher, G Dai, K Jannasch, C Dullin, F Alves, K Prenzel, WT Knoefel, NH Stoecklein; citation_volume=10; citation_publication_date=2011; citation_pages=30; citation_doi=10.1186/1476-4598-10-30; citation_id=CR17
citation_journal_title=J Immunol; citation_title=Cutting edge: the orphan chemokine receptor G protein-coupled receptor-2 (GPR-2, CCR10) binds the skin-associated chemokine CCL27 (CTACK/ALP/ILC); citation_author=B Homey, W Wang, H Soto, ME Buchanan, A Wiesenborn, D Catron, A Muller, TK McClanahan, MC Dieu-Nosjean, R Orozco; citation_volume=164; citation_publication_date=2000; citation_pages=3465-3470; citation_doi=10.4049/jimmunol.164.7.3465; citation_id=CR18
citation_journal_title=Genome Biol; citation_title=The chemokine and chemokine receptor superfamilies and their molecular evolution; citation_author=A Zlotnik, O Yoshie, H Nomiyama; citation_volume=7; citation_publication_date=2006; citation_pages=243; citation_doi=10.1186/gb-2006-7-12-243; citation_id=CR19
citation_journal_title=Proc Natl Acad Sci USA; citation_title=Monocyte chemoattractant protein 1 acts as a T-lymphocyte chemoattractant; citation_author=MW Carr, SJ Roth, E Luther, SS Rose, TA Springer; citation_volume=91; citation_publication_date=1994; citation_pages=3652-3656; citation_doi=10.1073/pnas.91.9.3652; citation_id=CR20
citation_journal_title=Am J Cancer Res; citation_title=Squamous cell carcinoma—similarities and differences among anatomical sites; citation_author=W Yan, II Wistuba, MR Emmert-Buck, HS Erickson; citation_volume=1; citation_publication_date=2011; citation_pages=275-300; citation_doi=10.1158/1538-7445.AM2011-275; citation_id=CR21
citation_journal_title=Pharmaceuticals (Basel); citation_title=Human antimicrobial peptides and proteins; citation_author=G Wang; citation_volume=7; citation_publication_date=2014; citation_pages=545-594; citation_doi=10.3390/ph7050545; citation_id=CR22
citation_journal_title=J Biol Chem; citation_title=Isolation and characterization of human beta-defensin-3, a novel human inducible peptide antibiotic; citation_author=J Harder, J Bartels, E Christophers, JM Schroder; citation_volume=276; citation_publication_date=2001; citation_pages=5707-5713; citation_doi=10.1074/jbc.M008557200; citation_id=CR23
citation_journal_title=J Biol Chem; citation_title=RNase 7, a novel innate immune defense antimicrobial protein of healthy human skin; citation_author=J Harder, JM Schroder; citation_volume=277; citation_publication_date=2002; citation_pages=46779-46784; citation_doi=10.1074/jbc.M207587200; citation_id=CR24
citation_journal_title=J Invest Dermatol; citation_title=Antimicrobial peptides human beta-defensins stimulate epidermal keratinocyte migration, proliferation and production of proinflammatory cytokines and chemokines; citation_author=F Niyonsaba, H Ushio, N Nakano, W Ng, K Sayama, K Hashimoto, I Nagaoka, K Okumura, H Ogawa; citation_volume=127; citation_publication_date=2007; citation_pages=594-604; citation_doi=10.1038/sj.jid.5700599; citation_id=CR25
citation_journal_title=Br J Dermatol; citation_title=Human papillomavirus-associated induction of human beta-defensins in anal intraepithelial neoplasia; citation_author=A Kreuter, M Skrygan, T Gambichler, NH Brockmeyer, M Stucker, C Herzler, A Potthoff, P Altmeyer, H Pfister, U Wieland; citation_volume=160; citation_publication_date=2009; citation_pages=1197-1205; citation_doi=10.1111/j.1365-2133.2009.09090.x; citation_id=CR26
citation_journal_title=Mol Cancer Ther; citation_title=Ingenol mebutate signals via PKC/MEK/ERK in keratinocytes and induces interleukin decoy receptors IL1R2 and IL13RA2; citation_author=SN Freiberger, PF Cheng, G Iotzova-Weiss, J Neu, Q Liu, P Dziunycz, JR Zibert, R Dummer, K Skak, MP Levesque, GF Hofbauer; citation_volume=14; citation_publication_date=2015; citation_pages=2132-2142; citation_doi=10.1158/1535-7163.MCT-15-0023-T; citation_id=CR27
citation_journal_title=J Cell Physiol; citation_title=Activation of protein kinase C inhibits human keratinocyte migration; citation_author=Y Ando, GS Lazarus, PJ Jensen; citation_volume=156; citation_publication_date=1993; citation_pages=487-496; citation_doi=10.1002/jcp.1041560308; citation_id=CR28
citation_journal_title=Cytokine; citation_title=Protein kinase c-dependent pathway is critical for the production of pro-inflammatory cytokines (TNF-alpha, IL-1beta, IL-6); citation_author=E Kontny, M Ziolkowska, A Ryzewska, W Maslinski; citation_volume=11; citation_publication_date=1999; citation_pages=839-848; citation_doi=10.1006/cyto.1998.0496; citation_id=CR29
citation_journal_title=Br J Dermatol; citation_title=Imiquimod: mode of action; citation_author=MP Schon, M Schon; citation_volume=157; citation_issue=Suppl 2; citation_publication_date=2007; citation_pages=8-13; citation_doi=10.1111/j.1365-2133.2007.08265.x; citation_id=CR30
citation_journal_title=J Natl Cancer Inst; citation_title=Photodynamic therapy; citation_author=TJ Dougherty, CJ Gomer, BW Henderson, G Jori, D Kessel, M Korbelik, J Moan, Q Peng; citation_volume=90; citation_publication_date=1998; citation_pages=889-905; citation_doi=10.1093/jnci/90.12.889; citation_id=CR31
citation_journal_title=Sci Rep; citation_title=The systemic response to Topical Aldara treatment is mediated through direct TLR7 stimulation as imiquimod enters the circulation; citation_author=L Nerurkar, A McColl, G Graham, J Cavanagh; citation_volume=7; citation_publication_date=2017; citation_pages=16570; citation_doi=10.1038/s41598-017-16707-5; citation_id=CR32
citation_journal_title=Br J Cancer; citation_title=Role of cytokines in photodynamic therapy-induced local and systemic inflammation; citation_author=SO Gollnick, SS Evans, H Baumann, B Owczarczak, P Maier, L Vaughan, WC Wang, E Unger, BW Henderson; citation_volume=88; citation_publication_date=2003; citation_pages=1772-1779; citation_doi=10.1038/sj.bjc.6600864; citation_id=CR33
citation_journal_title=Arch Dermatol; citation_title=A randomized, double-blind, vehicle-controlled study to assess 5% imiquimod cream for the treatment of multiple actinic keratoses; citation_author=E Stockfleth, T Meyer, B Benninghoff, S Salasche, L Papadopoulos, C Ulrich, E Christophers; citation_volume=138; citation_publication_date=2002; citation_pages=1498-1502; citation_doi=10.1001/archderm.138.11.1498; citation_id=CR34