Abstract
Chemical leukoderma is an acquired type of vitiligo that can be initiated by various exogenous chemicals such as hydroquinone (HQ), rhododendrol (RD), or 4-tertiary butyl phenol (4-TBP). Despite the importance of epidermal keratinocytes in diverse dermatological conditions, their toxicological role in chemical leukoderma is poorly understood. To elucidate their role in the pathogenesis of chemical leukoderma, genome-scale transcriptional analysis was performed in human epidermal keratinocytes (HEKs) treated with a sub-cytotoxic HQ concentration (10 µM). The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway-based functional enrichment analysis of HQ-induced differentially expressed genes (DEGs) revealed that HQ significantly upregulated DEGs related to the IL-17 signaling pathway and significantly downregulated DEGs associated with melanogenesis in HEKs. The meta-analysis between the HQ-induced and cytokine-induced transcriptional data (GSE53751) showed that 58 DEGs were commonly upregulated between HQ- and IL-17A-treated HEKs. Notably, the expression of IL36G was significantly increased in HEKs in response to both HQ and IL-17A. IL-36γ (2 µg/ml) directly inhibits melanin biosynthesis in cultured human epidermal melanocytes (HEMs) and downregulates the gene transcription of key enzymes in the melanogenesis pathway including TYR, DCT, and TYRP1. Moreover, IL-36γ autocrinally regulated keratinocyte function to produce the proinflammatory cytokines IL-36γ, IL-6, and CXCL8/IL-8 in a concentration-dependent manner, suggesting that IL-36γ may stimulate the amplification cycle of cutaneous inflammation. In this regard, hydroquinone-induced IL-36γ from human keratinocytes plays a pivotal role in the development of chemical leukoderma by autocrinally or paracrinally modulating the crosstalk between keratinocytes and melanocytes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ainscough JS, Macleod T, McGonagle D, Brakefield R, Baron JM, Alase A, Wittmann M, Stacey M (2017) Cathepsin S is the major activator of the psoriasis-associated proinflammatory cytokine IL-36γ. Proc Natl Acad Sci USA 114:E2748–E2757. https://doi.org/10.1073/pnas.1620954114
Ali S, Mohs A, Thomas M, Klare J, Ross R, Schmitz ML, Martin MU (2011) The dual function cytokine IL-33 interacts with the transcription factor NF-κB to dampen NF-κB-stimulated gene transcription. J Immunol 187:1609–1616. https://doi.org/10.4049/jimmunol.1003080
Ansel KM, Djuretic I, Tanasa B, Rao A (2006) Regulation of Th2 differentiation and Il4 locus accessibility. Annu Rev Immunol 24:607–656. https://doi.org/10.1146/annurev.immunol.23.021704.115821
Bassiouny DA, Shaker O (2011) Role of interleukin-17 in the pathogenesis of vitiligo. Clin Exp Dermatol 36:292–297. https://doi.org/10.1111/j.1365-2230.2010.03972.x
Boissy RE, Manga P (2004) On the etiology of contact/occupational vitiligo. Pigment Cell Res 17:208–214. https://doi.org/10.1111/j.1600-0749.2004.00130.x
Bonamonte D, Vestita M, Romita P, Filoni A, Foti C, Angelini G (2016) Chemical leukoderma. Dermatitis 27:90–99. https://doi.org/10.1097/DER.0000000000000167
Chitnis T, Salama AD, Grusby MJ, Sayegh MH, Khoury SJ (2004) Defining Th1 and Th2 immune responses in a reciprocal cytokine environment in vivo. J Immunol 172:4260–4265. https://doi.org/10.4049/jimmunol.172.7.4260
Choi H, Ahn S, Lee BG, Chang I, Hwang JS (2005) Inhibition of skin pigmentation by an extract of Lepidium apetalum and its possible implication in IL-6 mediated signaling. Pigment Cell Res 18:439–446. https://doi.org/10.1111/j.1600-0749.2005.00266.x
Choi H, Kim K, Han J, Choi H, Jin SH, Lee EK, Shin DW, Lee TR, Lee AY, Noh M (2012) Kojic acid-induced IL-6 production in human keratinocytes plays a role in its anti-melanogenic activity in skin. J Dermatol Sci 66:207–215. https://doi.org/10.1016/j.jdermsci.2012.03.002
Choi H, Choi H, Han J, Jin SH, Park JY, Shin DW, Lee TR, Kim K, Lee AY, Noh M (2013) IL-4 inhibits the melanogenesis of normal human melanocytes through the JAK2-STAT6 signaling pathway. J Invest Dermatol 133:528–536. https://doi.org/10.1038/jid.2012.331
Choy DF, Hart KM, Borthwick LA, Shikotra A, Nagarkar DR, Siddiqui S, Jia G, Ohri CM, Doran E, Vannella KM, Butler CA, Hargadon B, Sciurba JC, Gieseck RL, Thompson RW, White S, Abbas AR, Jackman J, Wu LC, Egen JG, Heaney LG, Ramalingam TR, Arron JR, Wynn TA, Bradding P (2015) TH2 and TH17 inflammatory pathways are reciprocally regulated in asthma. Sci Transl Med 7:301ra129. https://doi.org/10.1126/scitranslmed.aab3142
Chuaqui RF, Bonner RF, Best CJ, Gillespie JW, Flaig MJ, Hewitt SM, Phillips JL, Krizman DB, Tangrea MA, Ahram M, Linehan WM, Knezevic V, Emmert-Buck MR (2002) Post-analysis follow-up and validation of microarray experiments. Nat Genet 32:509–514. https://doi.org/10.1038/ng1034
Costin GE, Hearing VJ (2007) Human skin pigmentation: melanocytes modulate skin color in response to stress. FASEB J 21:976–994. https://doi.org/10.1096/fj.06-6649rev
Gabay C, Towne JE (2015) Regulation and function of interleukin-36 cytokines in homeostasis and pathological conditions. J Leukoc Biol 97:645–652. https://doi.org/10.1189/jlb.3RI1014-495R
Gor DO, Rose NR, Greenspan NS (2003) TH1-TH2: a procrustean paradigm. Nat Immunol 4:503–505. https://doi.org/10.1038/ni0603-503
Griffith JW, Sokol CL, Luster AD (2014) Chemokines and chemokine receptors: positioning cells for host defense and immunity. Annu Rev Immunol 32:659–702. https://doi.org/10.1146/annurev-immunol-032713-120145
Hariharan V, Klarquist J, Reust MJ, Koshoffer A, McKee MD, Boissy RE, Le Poole IC (2010) Monobenzyl ether of hydroquinone and 4-tertiary butyl phenol activate markedly different physiological responses in melanocytes: relevance to skin depigmentation. J Invest Dermatol 130:211–220. https://doi.org/10.1038/jid.2009.214
Henry CM, Sullivan GP, Clancy DM, Afonina IS, Kulms D, Martin SJ (2016) Neutrophil-derived proteases escalate inflammation through activation of IL-36 family cytokines. Cell Rep 14:708–722. https://doi.org/10.1016/j.celrep.2015.12.072
Imokawa G (2004) Autocrine and paracrine regulation of melanocytes in human skin and in pigmentary disorders. Pigment Cell Res 17:96–110. https://doi.org/10.1111/j.1600-0749.2003.00126.x
Ito S, Wakamatsu K (2018) Biochemical Mechanism of Rhododendrol-Induced Leukoderma. Int J Mol Sci 19:552. https://doi.org/10.3390/ijms19020552
Jin SH, Choi D, Chun YJ, Noh M (2014) Keratinocyte-derived IL-24 plays a role in the positive feedback regulation of epidermal inflammation in response to environmental and endogenous toxic stressors. Toxicol Appl Pharmacol 280:199–206. https://doi.org/10.1016/j.taap.2014.08.019
Johnston A, Xing X, Guzman AM, Riblett M, Loyd CM, Ward NL, Wohn C, Prens EP, Wang F, Maier LE, Kang S, Voorhees JJ, Elder JT, Gudjonsson JE (2011) IL-1F5, -F6, -F8, and -F9: a novel IL-1 family signaling system that is active in psoriasis and promotes keratinocyte antimicrobial peptide expression. J Immunol 186:2613–2622. https://doi.org/10.4049/jimmunol.1003162
Khan R, Gupta S, Sharma A (2012) Circulatory levels of T-cell cytokines (interleukin [IL]-2, IL-4, IL-17, and transforming growth factor-β) in patients with vitiligo. J Am Acad Dermatol 66:510–511. https://doi.org/10.1016/j.jaad.2011.07.018
Kim M, Baek HS, Lee M, Park H, Shin SS, Choi DW, Lim KM (2016) Rhododenol and raspberry ketone impair the normal proliferation of melanocytes through reactive oxygen species-dependent activation of GADD45. Toxicol In Vitro 32:339–346. https://doi.org/10.1016/j.tiv.2016.02.003
Kim SE, Lee CM, Kim YC (2017) Anti-melanogenic effect of Oenothera laciniata methanol extract in melan-a cells. Toxicol Res 33:55–62. https://doi.org/10.5487/TR.2017.33.1.055
Kooyers TJ, Westerhof W (2005) Toxicology and health risks of hydroquinone in skin lightening formulations. J Eur Acad Dermatol Venereol 20:7. https://doi.org/10.1111/j.1468-3083.2005.01218.x
Kotobuki Y, Tanemura A, Yang L, Itoi S, Wataya-Kaneda M, Murota H, Fujimoto M, Serada S, Naka T, Katayama I (2012) Dysregulation of melanocyte function by Th17-related cytokines: significance of Th17 cell infiltration in autoimmune vitiligo vulgaris. Pigment Cell Melanoma Res 25:219–230. https://doi.org/10.1111/j.1755-148X.2011.00945.x
Kwon OS, Kwon SJ, Kim JS, Lee G, Maeng HJ, Lee J, Hwang GS, Cha HJ, Chun KH (2018) Designing tyrosinase siRNAs by multiple prediction algorithms and evaluation of their anti-melanogenic effects. Biomol Ther 26:282–289. https://doi.org/10.4062/biomolther.2017.115
Lee CS, Joo YH, Baek HS, Park M, Kim JH, Shin HJ, Park NH, Lee JH, Park YH, Shin SS, Lee HK (2016a) Different effects of five depigmentary compounds, rhododendrol, raspberry ketone, monobenzone, rucinol and AP736 on melanogenesis and viability of human epidermal melanocytes. Exp Dermatol 25:44–49. https://doi.org/10.1111/exd.12871
Lee E, Kim HJ, Lee M, Jin SH, Hong SH, Ahn S, Kim SO, Shin DW, Lee ST, Noh M (2016b) Cystathionine metabolic enzymes play a role in the inflammation resolution of human keratinocytes in response to sub-cytotoxic formaldehyde exposure. Toxicol Appl Pharmacol 310:185–194. https://doi.org/10.1016/j.taap.2016.09.017
Lee M, Lee E, Jin SH, Ahn S, Kim SO, Kim J, Choi D, Lim KM, Lee ST, Noh M (2018) Leptin regulates the pro-inflammatory response in human epidermal keratinocytes. Arch Dermatol Res 310:351–362. https://doi.org/10.1007/s00403-018-1821-0
Li H, Hou L (2018) Regulation of melanocyte stem cell behavior by the niche microenvironment. Pigment Cell Melanoma Res 31:556–569. https://doi.org/10.1111/pcmr.12701
Morelli JG, Norris DA (1993) Influence of inflammatory mediators and cytokines on human melanocyte function. J Invest Dermatol 100:191S–195S. https://doi.org/10.1111/1523-1747.ep12465168
Mosher DB, Parrish JA, Fitzpatrick TB (1977) Monobenzylether of hydroquinone. A retrospective study of treatment of 18 vitiligo patients and a review of the literature. Br J Dermatol 97:669–679. https://doi.org/10.1111/j.1365-2133.1977.tb14275.x
Natarajan VT, Ganju P, Ramkumar A, Grover R, Gokhale RS (2014a) Multifaceted pathways protect human skin from UV radiation. Nat Chem Biol 10:542–551. https://doi.org/10.1038/nchembio.1548
Natarajan VT, Ganju P, Singh A, Vijayan V, Kirty K, Yadav S, Puntambekar S, Bajaj S, Dani PP, Kar HK, Gadgil CJ, Natarajan K, Rani R, Gokhale RS (2014b) IFN-γ signaling maintains skin pigmentation homeostasis through regulation of melanosome maturation. Proc Natl Acad Sci USA 111:2301–2306. https://doi.org/10.1073/pnas.1304988111
Pfaff CM, Marquardt Y, Fietkau K, Baron JM, Lüscher B (2017) The psoriasis-associated IL-17A induces and cooperates with IL-36 cytokines to control keratinocyte differentiation and function. Sci Rep 7:15631. https://doi.org/10.1038/s41598-017-15892-7
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45. https://doi.org/10.1093/nar/29.9.e45
Picardo M, Dell’Anna ML, Ezzedine K, Hamzavi I, Harris JE, Parsad D, Taieb A (2015) Vitiligo. Nat Rev Dis Primers 1:15011. https://doi.org/10.1038/nrdp.2015.11
Rashighi M, Agarwal P, Richmond JM, Harris TH, Dresser K, Su MW, Zhou Y, Deng A, Hunter CA, Luster AD, Harris JE (2014) CXCL10 is critical for the progression and maintenance of depigmentation in a mouse model of vitiligo. Sci Transl Med 6:22323. https://doi.org/10.1126/scitranslmed.3007811
Regazzetti C, Joly F, Marty C, Rivier M, Mehul B, Reiniche P, Mounier C, Rival Y, Piwnica D, Cavalié M, Chignon-Sicard B, Ballotti R, Voegel J, Passeron T (2015) Transcriptional analysis of vitiligo skin reveals the alteration of WNT pathway: a promising target for repigmenting vitiligo patients. J Invest Dermatol. 135:3105–3114. https://doi.org/10.1038/jid.2015.335
Saldana-Caboverde A, Kos L (2010) Roles of endothelin signaling in melanocyte development and melanoma. Pigment Cell Melanoma Res 23:160–170. https://doi.org/10.1111/j.1755-148X.2010.00678.x
Schmitz J, Owyang A, Oldham E, Song Y, Murphy E, McClanahan TK, Zurawski G, Moshrefi M, Qin J, Li X, Gorman DM, Bazan JF, Kastelein RA (2005) IL-33, an interleukin-1-like cytokine that signals via the IL-1 receptor-related protein ST2 and induces T helper type 2-associated cytokines. Immunity 23:479–490. https://doi.org/10.1016/j.immuni.2005.09.015
Schulte G, Bryja V (2007) The Frizzled family of unconventional G-protein-coupled receptors. Trends Pharmacol Sci 28:518–525. https://doi.org/10.1016/j.tips.2007.09.001
Singh A, Gotherwal V, Junni P, Vijayan V, Tiwari M, Ganju P, Kumar A, Sharma P, Fatima T, Gupta A, Holla A, Kar HK, Khanna S, Thukral L, Malik G, Natarajan K, Gadgil CJ, Lahesmaa R, Natarajan VT, Rani R, Gokhale RS (2017) Mapping architectural and transcriptional alterations in non-lesional and lesional epidermis in vitiligo. Sci Rep 7:9860. https://doi.org/10.1038/s41598-017-10253-w
Slominski A, Tobin DJ, Shibahara S, Wortsman J (2004) Melanin pigmentation in mammalian skin and its hormonal regulation. Physiol Rev 84:1155–1228. https://doi.org/10.1152/physrev.00044.2003
Speeckaert R, van Geel N (2017) Vitiligo: an update on pathophysiology and treatment options. Am J Clin Dermatol 18:733–744. https://doi.org/10.1007/s40257-017-0298-5
Swindell WR, Beamer MA, Sarkar MK, Loftus S, Fullmer J, Xing X, Ward NL, Tsoi LC, Kahlenberg MJ, Liang Y, Gudjonsson JE (2018) RNA-Seq analysis of IL-1B and IL-36 responses in epidermal keratinocytes identifies a shared MyD88-dependent gene signature. Front Immunol 9:80. https://doi.org/10.3389/fimmu.2018.00080
Swope VB, Abdel-Malek Z, Kassem LM, Nordlund JJ (1991) Interleukins 1 alpha and 6 and tumor necrosis factor-alpha are paracrine inhibitors of human melanocyte proliferation and melanogenesis. J Invest Dermatol 96:180–185. https://doi.org/10.1111/1523-1747.ep12460991
Teunissen MB, Koomen CW, de Waal Malefyt R, Wierenga EA, Bos JD (1998) Interleukin-17 and interferon-gamma synergize in the enhancement of proinflammatory cytokine production by human keratinocytes. J Invest Dermatol 111:645–649. https://doi.org/10.1046/j.1523-1747.1998.00347.x
Tokura Y, Fujiyama T, Ikeya S, Tatsuno K, Aoshima M, Kasuya A, Ito T (2015) Biochemical, cytological, and immunological mechanisms of rhododendrol-induced leukoderma. J Dermatol Sci 77:146–149. https://doi.org/10.1016/j.jdermsci.2015.02.001
Toosi S, Orlow SJ, Manga P (2012) Vitiligo-inducing phenols activate the unfolded protein response in melanocytes resulting in upregulation of IL6 and IL8. J Invest Dermatol 132:2601–2609. https://doi.org/10.1038/jid.2012.181
US FDA. Department of Health and Human Services (2009) Supporting information for toxicological evaluation by the national toxicology program—hydroquinone (CAS 123-31-9). https://ntp.niehs.nih.gov/ntp/noms/support_docs/hydroquinone_may2009.pdf
Wang CQ, Cruz-Inigo AE, Fuentes-Duculan J, Moussai D, Gulati N, Sullivan-Whalen M, Gilleaudeau P, Cohen JA, Krueger JG (2011) Th17 cells and activated dendritic cells are increased in vitiligo lesions. PLoS ONE 6:e18907. https://doi.org/10.1371/journal.pone.0018907
Wilson CB, Rowell E, Sekimata M (2009) Epigenetic control of T-helper-cell differentiation. Nat Rev Immunol 9:91–105. https://doi.org/10.1038/nri2487
Yamada T, Hasegawa S, Inoue Y, Date Y, Yamamoto N, Mizutani H, Nakata S, Matsunaga K, Akamatsu H (2013) Wnt/β-catenin and kit signaling sequentially regulate melanocyte stem cell differentiation in UVB-induced epidermal pigmentation. J Invest Dermatol 133:2753–2762. https://doi.org/10.1038/jid.2013.235
Yang F, Sarangarajan R, Le Poole IC, Medrano EE, Boissy RE (2000) The cytotoxicity and apoptosis induced by 4-tertiary butylphenol in human melanocytes are independent of tyrosinase activity. J Invest Dermatol 114:157–164. https://doi.org/10.1046/j.1523-1747.2000.00836.x
Yang L, Yang F, Wataya-Kaneda M, Tanemura A, Tsuruta D, Katayama I (2015) 4-(4-hydroroxyphenyl)-2-butanol (rhododendrol) activates the autophagy-lysosome pathway in melanocytes: insights into the mechanisms of rhododendrol-induced leukoderma. J Dermatol Sci 77:182–185. https://doi.org/10.1016/j.jdermsci.2015.01.006
Acknowledgements
This study was partly supported by the MRC Grant of the National Research Foundation in Korea (NRF-2018R1A5A2024425) and a National Research Foundation of Korea (NRF) Grant (2015R1A2A2A01008408).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
Hyunjung Choi and Hyoung-June Kim are employees of AmorePacific. The other authors have no conflicts of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Pyo, J.J., Ahn, S., Jin, S.H. et al. Keratinocyte-derived IL-36γ plays a role in hydroquinone-induced chemical leukoderma through inhibition of melanogenesis in human epidermal melanocytes. Arch Toxicol 93, 2307–2320 (2019). https://doi.org/10.1007/s00204-019-02506-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-019-02506-6