Animal Bioscience

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
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Exosomal miR-222-3p derived from dermal papilla cells inhibits melanogenesis in melanocytes by targeting SOX10 in rabbits

  • Yang Chen (College of Animal Science and Technology, Yangzhou University) ;
  • Tingting Lu (College of Animal Science and Technology, Yangzhou University) ;
  • Yingying Dai (College of Animal Science and Technology, Yangzhou University) ;
  • Yu Xue (College of Animal Science and Technology, Yangzhou University) ;
  • Bohao Zhao (College of Animal Science and Technology, Yangzhou University) ;
  • Xinsheng Wu (College of Animal Science and Technology, Yangzhou University)
  • Received : 2024.03.26
  • Accepted : 2024.08.16
  • Published : 2025.02.01
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Abstract

Objective: Dermal papilla cells (DPCs) play a pivotal role in hair follicle development and can modulate melanogenesis in melanocytes (MCs) through their microenvironment. Our previous studies have demonstrated that the levels of exosomal miR-222-3p derived from DPCs of white Rex rabbits are significantly higher than those of black Rex rabbits. However, the specific role and underlying molecular mechanisms of exosomal miR-222-3p in melanogenesis remain elusive. Methods: DPCs and MCs were isolated from hair follicles of Rex rabbits and identified using western blotting (WB) and immunofluorescent staining. Exosomes derived from DPCs (DPCs-exos) were characterized using nanoparticle tracking analysis, transmission electron microscopy, and WB. To investigate cell-cell crosstalk mediated by exosomes, MCs were co-cultured with CM-Dil-labeled DPCs-exos. The expression of miR-222-3p in skin tissue and exosomes was quantitatively assessed using quantitative real-time polymerase chain reaction. The transmission of DPCs-secreted exosomal miR-222-3p to MCs was demonstrated using Cy3-labeled miR-222-3p in conjunction with transwell assays. The impact of miR-222-3p on melanin synthesis was evaluated using the NaOH method, cell counting kit-8, and annexin V-fluorescein isothiocyanate/propidium iodide assays. Sex determining region Y-box 10 (SOX10), a potential target gene regulated by miR-222-3p, was validated using a dual-luciferase reporter assay, site-specific mutation, and WB. Results: Increased levels of miR-222-3p were observed in the skin and DPCs-exos of white Rex rabbits compared to those of black Rex rabbits. Effective internalization of CM-Dillabeled DPCs-exos by MCs was observed. Furthermore, exosomal miR-222-3p derived from DPCs was transferred to MCs. Functionally, miR-222-3p significantly inhibited MCs proliferation, induced apoptosis and inhibited melanin synthesis. SOX10 was confirmed as a direct target of miR-222-3p in this regulatory cascade. Conclusion: The findings demonstrate that exosomal miR-222-3p, derived from DPCs, suppresses melanogenesis in MCs by targeting SOX10, thus unveiling a novel mechanism of exosome involvement in melanogenesis.

Keywords

Acknowledgement

This work was supported by National Natural Science Grant Nos. 32172722, the Qing Lan Project of Yangzhou University, China Agriculture Research System of MOF and MARA (CARS-43-A-1), and the Natural Science Foundation of Jiangsu Province (BK20231332).

References

  1. Ali SA, Naaz I. Biochemical aspects of mammalian melanocytes and the emerging role of melanocyte stem cells in dermatological therapies. Int J Health Sci 2018;12:69-76.
  2. Yamaguchi Y, Hearing VJ. Melanocytes and their diseases. Cold Spring Harb Perspect Med 2014;4:a017046. https://doi.org/10.1101/cshperspect.a017046
  3. Lim J, Ng KJ, Clavel C. Dermal papilla regulation of hair growth and pigmentation. Adv Stem Cell Niches 2019;3: 115-38. https://doi.org/10.1016/bs.asn.2019.06.002
  4. Slominski A, Wortsman J, Plonka PM, Schallreuter KU, Paus R, Tobin D. Hair follicle pigmentation. J Invest Dermatol 2005;124:13-21. https://doi.org/10.1111/j.0022-202X.2004.23528.x
  5. Simons M, Raposo G. Exosomes - vesicular carriers for intercellular communication. Curr Opin Cell Biol 2009;21: 575-81. https://doi.org/10.1016/j.ceb.2009.03.007
  6. Raposo G, Stoorvogel W. Extracellular vesicles: exosomes, microvesicles, and friends. J Cell Biol 2013;200:373-83. https://doi.org/10.1083/jcb.201211138
  7. Gross JC, Chaudhary V, Bartscherer K, Boutros M. Active Wnt proteins are secreted on exosomes. Nat Cell Biol 2012; 14:1036-45. https://doi.org/10.1038/ncb2574
  8. Rajendran RL, Gangadaran P, Bak SS, et al. Extracellular vesicles derived from MSCs activates dermal papilla cell in vitro and promotes hair follicle conversion from telogen to anagen in mice. Sci Rep 2017;7:15560. https://doi.org/10.1038/s41598-017-15505-3
  9. Riche A, Aberdam E, Marchand L, et al. Extracellular vesicles from activated dermal fibroblasts stimulate hair follicle growth through dermal papilla-secreted norrin. Stem Cells 2019;37: 1166-75. https://doi.org/10.1002/stem.3043
  10. Zhou L, Wang H, Jing J, Yu LJ, Wu XJ, Luet ZF. Regulation of hair follicle development by exosomes derived from dermal papilla cells. Biochem Biophys Res Commun 2018;500:325-32. https://doi.org/10.1016/j.bbrc.2018.04.067
  11. Hu S, Li Z, Lutz H, et al. Dermal exosomes containing miR218-5p promote hair regeneration by regulating beta-catenin signaling. Sci Adv 2020;6:eaba1685. https://doi.org/10.1126/sciadv.aba1685
  12. Chen Y, Huang J, Liu Z, et al. miR-140-5p in small extracellular vesicles from human papilla cells stimulates hair growth by promoting proliferation of outer root sheath and hair matrix cells. Front Cell Dev Biol 2020;8:593638. https://doi.org/10.3389/fcell.2020.593638
  13. Kwack MH, Seo CH, Gangadaran P, et al. Exosomes derived from human dermal papilla cells promote hair growth in cultured human hair follicles and augment the hair‐inductive capacity of cultured dermal papilla spheres. Exp Dermatol 2019;28:854-7. https://doi.org/10.1111/exd.13927
  14. Carrasco E, Soto-Heredero G, Mittelbrunn M. The role of extracellular vesicles in cutaneous remodeling and hair follicle dynamics. Int J Mol Sci 2019;20:2758. https://doi.org/10.3390/ijms20112758
  15. Tarafder AK, Bolasco G, Correia MS, et al. Rab11b mediates melanin transfer between donor melanocytes and acceptor keratinocytes via coupled exo/endocytosis. J Invest Dermatol 2014;134:1056-66. https://doi.org/10.1038/jid.2013.432
  16. Liu Y, Xue L, Gao H, et al. Exosomal miRNA derived from keratinocytes regulates pigmentation in melanocytes. J Dermatol Sci 2019;93:159-67. https://doi.org/10.1016/j.jdermsci.2019.02.001
  17. Lo Cicero A, Delevoye C, Gilles-Marsens F, et al. Exosomes released by keratinocytes modulate melanocyte pigmentation. Nat Commun 2015;6:7506. https://doi.org/10.1038/ncomms8506
  18. Yoon YJ, Kim OY, Gho YS. Extracellular vesicles as emerging intercellular communicasomes. BMB Rep 2014;47:531-9. https://doi.org/10.5483/BMBRep.2014.47.10.164
  19. Kim NH, Choi SH, Kim CH, Lee CH, Lee TR, Lee AY. Reduced MiR-675 in exosome in H19 RNA-related melanogenesis via MITF as a direct target. J Invest Dermatol 2014;134: 1075-82. https://doi.org/10.1038/jid.2013.478
  20. Pfeffer SR, Grossmann KF, Cassidy PB, et al. Detection of exosomal miRNAs in the plasma of melanoma patients. J Clin Med 2015;4:2012-27. https://doi.org/10.3390/jcm4121957
  21. Tang YQ, Cao YM. Curcumin inhibits the growth and metastasis of melanoma via miR-222-3p/SOX10/Notch Axis. Dis Markers 2022;2022:3129781. https://doi.org/10.1155/2022/3129781
  22. Isacescu E, Chiroi P, Zanoaga O, et al. Melanoma cellular signaling transduction pathways targeted by polyphenols action mechanisms. Antioxidants 2023;12:407. https://doi.org/10.3390/antiox12020407
  23. Levstek T, Vujkovac B, Vujkovac AC, Podkrajsek KT. Urinaryderived extracellular vesicles reveal a distinct microRNA signature associated with the development and progression of Fabry nephropathy. Front Med 2023;10:1143905. https://doi.org/10.3389/fmed.2023.1143905
  24. Zhao B, Chen Y, Mu L, Hu S, Wu X. Identification and profiling of microRNA between back and belly skin in rex rabbits (Oryctolagus cuniculus). World Rabbit Sci 2018;26: 179-90. https://doi.org/10.4995/wrs.2018.7058
  25. Baker R, Thornton MJ. Isolation of epidermal and hair follicle melanocytes. Methods Mol Biol 2020;2154:23-32. https://doi.org/10.1007/978-1-0716-0648-3_3
  26. Fan RR, Liu XM, Liu FL, et al. Melanocytes derived from mouse hair follicles: A novel study model to assess pigmentation disorders. Pathol Res Pract 2020;216:153224. https://doi.org/10.1016/j.prp.2020.153224
  27. Livak KJ, Schmittgen TD. Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT method. Methods 2001;25:402-8. https://doi.org/10.1006/meth.2001.1262
  28. Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative CT method. Nat Protoc 2008;3:1101-8. https://doi.org/10.1038/nprot.2008.73
  29. Alexeev V, Yoon K. Distinctive role of the cKit receptor tyrosine kinase signaling in mammalian melanocytes. J Invest Dermatol 2006;126:1102-10. https://doi.org/10.1038/sj.jid.5700125
  30. Botchkareva NV, Khlgatian M, Longley BJ, Botchkarev VA, Gilchrest BA. SCF/c-kit signaling is required for cyclic regeneration of the hair pigmentation unit. FASEB J 2001; 15:645-58. https://doi.org/10.1096/fj.00-0368com
  31. Baynash AG, Hosoda K, Giaid A, et al. Interaction of endothelin-3 with endothelin-B receptor is essential for development of epidermal melanocytes and enteric neurons. Cell 1994;79:1277-85. https://doi.org/10.1016/0092-8674(94) 90018-3
  32. Garcia RJ, Ittah A, Mirabal S, et al. Endothelin 3 induces skin pigmentation in a keratin-driven inducible mouse model. J Invest Dermatol 2008;128:131-42. https://doi.org/10.1038/sj.jid.5700948
  33. Lahav R, Ziller C, Dupin E, Le Douarin NM. Endothelin 3 promotes neural crest cell proliferation and mediates a vast increase in melanocyte number in culture. Proc Natl Acad Sci USA 1996;93:3892-7. https://doi.org/10.1073/pnas.93.9.3892
  34. Ideta R, Soma T, Tsunenaga M, Ifuku O. Cultured human dermal papilla cells secrete a chemotactic factor for melanocytes. J Dermatol Sci 2002;28:48-59. https://doi.org/10.1016/s0923-1811(01)00145-1
  35. Vrieling H, Duhl D, Millar SE, Miller KA, Barsh GS. Differences in dorsal and ventral pigmentation result from regional expression of the mouse agouti gene. Proc Natl Acad Sci USA 1994;91:5667-71. https://doi.org/10.1073/pnas.91.12.5667
  36. Hoekstra HE. Genetics, development and evolution of adaptive pigmentation in vertebrates. Heredity 2006;97:222-34. https://doi.org/10.1038/sj.hdy.6800861
  37. Balafa C, Smith‐Thomas L, Phillips J, et al. Dopa oxidase activity in the hair, skin and ocular melanocytes is increased in the presence of stressed fibroblasts. Exp Dermatol 2005; 14:363-72. https://doi.org/10.1111/j.0906-6705.2005.00287.x
  38. Liu Y, Xue L, Gao H, et al. Exosomal miRNA derived from keratinocytes regulates pigmentation in melanocytes. J Dermatol Sci 2019;93:159-67. https://doi.org/10.1016/j.jdermsci.2019.02.001
  39. Zhao CS, Wang DL, Wang X, et al. Down-regulation of exosomal miR-200c derived from keratinocytes in vitiligo lesions suppresses melanogenesis. J Cell Mol Med 2020;24: 12164-75. https://doi.org/10.1111/jcmm.15864
  40. Wang XY, Guan XH, Yu ZP, et al. Human amniotic stem cells-derived exosmal miR-181a-5p and miR-199a inhibit melanogenesis and promote melanosome degradation in skin hyperpigmentation, respectively. Stem Cell Res Ther 2021;12:501. https://doi.org/10.1186/s13287-021-02570-9
  41. Bae IS, Kim SH. milk exosome-derived MicroRNA-2478 suppresses melanogenesis through the Akt-GSK3 beta pathway. Cells 2021;10:2848. https://doi.org/10.3390/cells10112848
  42. Mueller DW, Rehli M, Bosserhoff AK. miRNA expression profiling in melanocytes and melanoma cell lines reveals miRNAs associated with formation and progression of malignant melanoma. J Invest Dermatol 2009;129:1740-51. https://doi.org/10.1038/jid.2008.452
  43. Nonaka D, Chiriboga L, Rubin BP. Sox10: a pan-schwannian and melanocytic marker. Am J Surg Pathol 2008;32:1291-8. https://doi.org/10.1097/PAS.0b013e3181658c14
  44. Thway K, Hamarneh W, Miah AB, Fisher C. Malignant peripheral nerve sheath tumor with rhabdomyosarcomatous and glandular elements: rare epithelial differentiation in a triton tumor. Int J Surg Pathol 2015;23:377-83. https://doi.org/10.1177/1066896915583996
  45. Aoki Y, Saint-Germain N, Gyda M, et al. Sox10 regulates the development of neural crest-derived melanocytes in Xenopus. Dev Biol 2003;259:19-33. https://doi.org/10.1016/s0012-1606(03)00161-1
  46. Cook AL, Smith AG, Smit DJ, Leonard JH, Sturm RA. Coexpression of SOX9 and SOX10 during melanocytic differentiation in vitro. Exp Cell Res 2005;308:222-35. https://doi.org/10.1016/j.yexcr.2005.04.019
  47. Hou L, Arnheiter H, Pavan WJ. Interspecies difference in the regulation of melanocyte development by SOX10 and MITF. Proc Natl Acad Sci USA 2006;103:9081-5. https://doi.org/10.1073/pnas.0603114103
  48. Britsch S, Goerich DE, Riethmacher D, et al. The transcription factor Sox10 is a key regulator of peripheral glial development. Genes Dev 2001;15:66-78. https://doi.org/10.1101/gad.186601