Dermal fibroblast expression of stromal cellderived factor-1 (SDF-1) promotes epidermal keratinocyte proliferation in normal and diseased skin

Abstract

Stromal cells provide a crucial microenvironment for overlying epithelium. Hereweinvestigated the expression and function of a stromal cell-specific protein, stromal cell-derived factor-1 (SDF-1), in normal human skin and in the tissues of diseased skin. Immunohistology and laser capture microdissection (LCM)-coupled quantitative realtime RT-PCR revealed that SDF-1 is constitutively and predominantly expressed in dermal stromal cells in normalhuman skin in vivo. To our surprise, an extremely high level of SDF-1 transcription was observed in the dermis of normal human skin in vivo, evidenced by much higher mRNA expression level than type I collagen, the most abundant and highly expressed protein in human skin. SDF-1 was also upregulated in the tissues of manyhuman skin disorders including psoriasis, basal cell carcinoma (BCC), and squamous cell carcinoma (SCC). Double immunostaining for SDF-1 and HSP47 (heat shock protein 47), a marker of fibroblasts, revealed that fibroblasts were the major source of stroma-cell-derived SDF-1 in both normal and diseased skin. Functionally, SDF-1 activates the ERK (extracellular-signal-regulated kinases) pathway and functions as a mitogen to stimulate epidermal keratinocyte proliferation. Both overexpression of SDF-1 in dermal fibroblasts and treatment with rhSDF-1 to the skin equivalent cultures significantly increased the number of keratinocyte layers and epidermal thickness. Conversely, the stimulative function of SDF-1 on keratinocyte proliferation was nearly completely eliminated by interfering with CXCR4, a specific receptor of SDF-1, or by knock-down of SDF-1 in fibroblasts. Our data reveal that extremely high levels of SDF-1 provide a crucial microenvironment for epidermal keratinocyte proliferation in both physiologic and pathologic skin conditions.

Key words： SDF-1; dermal fibroblast; keratinocyte; proliferation; skin cancer; psoriasis

References

1 Bhowmick NA, Neilson EG, Moses HL (2004) Stromal fibroblasts in cancer initiation and progression. Nature 432:332–337

2 Blanpain C, Fuchs E (2009) Epidermal homeostasis: a balancing act of stem cells in the skin. Nat Rev 10:207–217

3 Boelsma E, Verhoeven MC, Ponec M (1999) Reconstruction of a human skin equivalent using a spontaneously transformed keratinocyte cell line (HaCaT). J Investig Dermatol 112:489–498

4 Bollag WB, Hill WD (2013) CXCR4 in epidermal keratinocytes: crosstalk within the skin. J Investig Dermatol 133:2505–2508

5 Byrne SN, Sarchio SN (2014) AMD3100 protects from UV-induced skin cancer. Oncoimmunology3:e27562

6 Chambard JC, Lefloch R, Pouyssegur J, Lenormand P (2007) ERK implication in cell cycle regulation. Biochimica et biophysica acta 1773:1299–1310

7 Chen GS, Yu HS, Lan CC, Chow KC, Lin TY, Kok LF, Lu MP, Liu CH, Wu MT (2006) CXC chemokine receptor CXCR4 expression enhances tumorigenesis and angiogenesis of basal cell carcinoma. Br J Dermatol 154:910–918

8 Chen SJ, Nakahara T, Kido M, Takahara M, Uchi H, Takeuchi S, Dugu L, Tu YT, Moroi Y, Furue M (2009) Stromal cell-derived factor 1 expression in various skin tumours. Br J Dermatol 160:710–713

9 Chu CY, Cha ST, Lin WC, Lu PH, Tan CT, Chang CC, Lin BR, Jee SH, Kuo ML (2009) Stromal cell-derived factor-1alpha (SDF-1alpha/CXCL12)-enhanced angiogenesis of human basal cell carcinoma cells involves ERK1/2-NF-kappaB/interleukin-6 pathway. Carcinogenesis 30:205–213

10 Donjacour AA, Cunha GR (1991) Stromal regulation of epithelial function. Cancer Treat Res 53:335–364

11 Duda DG, Kozin SV, Kirkpatrick ND, Xu L, Fukumura D, Jain RK (2011) CXCL12 (SDF1alpha)-CXCR4/CXCR7 pathway inhibition: an emerging sensitizer for anticancer therapies? Clin Cancer Res 17:2074–2080

12 Eckert RL, Efimova T, Dashti SR, Balasubramanian S, Deucher A, Crish JF, Sturniolo M, Bone F (2002) Keratinocyte survival, differentiation, and death: many roads lead to mitogen-activated protein kinase. J Investig Dermatol 7:36–40

13 El Ghalbzouri A, Lamme E,Ponec M (2002) Crucial role of fibroblasts in regulating epidermal morphogenesis. Cell Tissue Res 310:189–199

14 Farage MA, Miller KW, Maibach HI, SpringerLink (Online Service) (2010) Textbook of aging skin. Springer, Berlin

15 Fisher GJ, Esmann J, Griffiths CE, Talwar HS, Duell EA, Hammerberg C, Elder JT, Finkel LJ, Karabin GD, Nickoloff BJ (1991) Cellular, immunologic and biochemical characterization of topical retinoic acid-treated human skin. J Invest Dermatol 96:699–707

16 Fisher GJ, Varani J, Voorhees JJ (2008) Looking older: fibroblast collapse and therapeutic implications. Arch Dermatol 144:666–672

17 Fisher GJ, Quan T, Purohit T, Shao Y, Cho MK, He T, Varani J, Kang S, Voorhees JJ (2009) Collagen fragmentation promotes oxidative stress and elevates matrix metalloproteinase-1 in fibroblasts in aged human skin. The American journal of pathology. 174:101–114

18 Gallagher KA, Liu ZJ, Xiao M, Chen H, Goldstein LJ, Buerk DG, Nedeau A, Thom SR, Velazquez OC (2007) Diabetic impairments in NO-mediated endothelial progenitor cell mobilization and homing are reversed by hyperoxia and SDF-1 alpha. J Clin Investig 117:1249–1259

19 Henderson PW, Singh SP, Krijgh DD, Yamamoto M, Rafii DC, Sung JJ, Rafii S, Rabbany SY, Spector JA (2011) Stromal-derived factor-1 delivered via hydrogel drug-delivery vehicle accelerates wound healing in vivo. Wound Repair Regener 19:420–425

20 Hu TH, Yao Y, Yu S, Han LL, Wang WJ, Guo H, Tian T, Ruan ZP, Kang XM, Wang J, Wang SH, Nan KJ (2014) SDF-1/CXCR4 promotes epithelial-mesenchymal transition and progression of colorectal cancer by activation of the Wnt/beta-catenin signaling pathway. Cancer Lett 354:417–426

21 Jost M, Huggett TM, Kari C, Boise LH, Rodeck U (2001) Epidermal growth factor receptor-dependent control of keratinocyte survival and Bcl-xL expression through a MEK-dependent pathway. J Biol Chem 276:6320–6326

22 Kalatskaya I, Berchiche YA, Gravel S, Limberg BJ, Rosenbaum JS, Heveker N (2009) AMD3100 is a CXCR7 ligand with allosteric agonist properties. Mol Pharm 75:1240–1247

23 Kijima T, Maulik G, Ma PC, Tibaldi EV, Turner RE, Rollins B, Sattler M, Johnson BE, Salgia R (2002) Regulation of cellular proliferation, cytoskeletal function, and signal transduction through CXCR4 and c-Kit in small cell lung cancer cells. Cancer Res 62:6304–6311

24 Koster MI (2009) Making an epidermis. Ann N Y Acad Sci 1170:7–10

25 Kryczek I, Wei S, Keller E, Liu R, Zou W (2007) Stroma-derived factor (SDF-1/CXCL12) and human tumor pathogenesis. Am J Physiol 292:C987–995

26 Lapidot T, Dar A, Kollet O (2005) How do stem cells find their way home? Blood 106:1901–1910

27 Li X, Ma Q, Xu Q, Liu H, Lei J, Duan W, Bhat K, Wang F, Wu E, Wang Z (2012) SDF-1/CXCR4 signaling induces pancreatic cancer cell invasion and epithelial-mesenchymal transition in vitro through noncanonical activation of Hedgehog pathway. Cancer Lett 322:169–176

28 Li X, Li P, Chang Y, Xu Q, Wu Z, Ma Q, Wang Z (2014) The SDF-1/CXCR4 axis induces epithelial-mesenchymal transition in hepatocellular carcinoma. Mol Cell Biochem 392:77–84

29 Lin Q, Wesson RN, Maeda H, Wang Y, Cui Z, Liu JO, Cameron AM, Gao B, Montgomery RA, Williams GM, Sun Z (2014) Pharmacological mobilization of endogenous stem cells significantly promotes skin regeneration after full-thickness excision: the synergistic activity of AMD3100 and tacrolimus. J Investig Dermatol 134:2458–2468

30 Luker KE, Luker GD (2006) Functions of CXCL12 and CXCR4 in breast cancer. Cancer letters. 238:30–41

31 Maas-Szabowski N, Shimotoyodome A, Fusenig NE (1999) Keratinocyte growth regulation in fibroblast cocultures via a double paracrine mechanism. J Cell Sci 112(Pt 12):1843–1853

32 Maksym RB, Tarnowski M, Grymula K, Tarnowska J, Wysoczynski M, Liu R, Czerny B, Ratajczak J, Kucia M, Ratajczak, MZ (2009) The role of stromal-derived factor-1- CXCR7 axis in development and cancer. Eur J Pharmacol 625:31–40

33 Mine S, Fortunel NO, Pageon H, Asselineau D (2008) Aging alters functionally human dermal papillary fibroblasts but not reticular fibroblasts: a new view of skin morphogenesis and aging. PloS One 3:e4066

34 Mueller MM, Fusenig NE (2004) Friends or foes- bipolar effects of the tumour stroma in cancer. Nature reviews. Cancer. 4:839–849

35 Nie Y,Han YC, Zou YR(2008)CXCR4is required for the quiescence of primitive hematopoietic cells. J Exp Med 205:777–783

36 Nishimura Y, Ii M, Qin G, Hamada H, Asai J, Takenaka H, Sekiguchi H, Renault MA, Jujo K, Katoh N, Kishimoto S, Ito A, Kamide C, Kenny J, Millay M, Misener S, Thorne T, Losordo DW (2012) CXCR4 antagonist AMD3100 accelerates impaired wound healing in diabetic mice. J Investig Dermatol 132:711–720

37 Onoue T, Uchida D, Begum NM, Tomizuka Y, Yoshida H, Sato M (2006) Epithelial-mesenchymal transition induced by the stromal cell-derived factor-1/CXCR4 system in oral squamous cell carcinoma cells. Int J Oncol 29:1133–1138

38 Orimo A, Gupta PB, Sgroi DC, Arenzana-Seisdedos F, Delaunay T, Naeem R, Carey VJ, Richardson AL, Weinberg RA (2005) Stromal fibroblasts present in invasive human breast carcinomas promote tumor growth and angiogenesis through elevated SDF-1/CXCL12 secretion. Cell 121:335–348

39 Pablos JL, Amara A, Bouloc A, Santiago B, Caruz A, Galindo M, Delaunay T, Virelizier JL, Arenzana-Seisdedos F (1999) Stromalcell derived factor is expressed by dendritic cells and endothelium in human skin. Am J Pathol 155:1577–1586

40 Pickard A, Cichon AC, Menges C, Patel D, McCance DJ (2012) Regulation of epithelial differentiation and proliferation by the stroma: a role for the retinoblastoma protein. J Investig Dermatol 132:2691–2699

41 Potten CS (1981) Cell replacement in epidermis (keratopoiesis) via discrete units of proliferation. Int Rev Cytol 69:271–318

42 Qin Z, Fisher GJ, Quan T (2013) CCN1 domain-specific stimulation of matrix metalloproteinase-1 expression through alphaVbeta3 integrin in human skin fibroblasts. J Biol Chem 288:12386–12394

43 Quail DF, Joyce JA (2013) Microenvironmental regulation of tumor progression and metastasis. Nat Med 19:1423–1437

44 Quan T, He T, Voorhees JJ, Fisher GJ (2001) Ultraviolet irradiation blocks cellular responses to transforming growth factor-β by down-regulating its type-II receptor and inducing Smad7. J Biol Chem 276:26349–26356

45 Quan T, Qin Z, Xia W, Shao Y, Voorhees JJ, Fisher GJ (2009) Matrixdegrading metalloproteinases in photoaging. J Investig Dermatol Symp Proc 14:20–24

46 Quan T, Shao Y, He T, Voorhees JJ, Fisher GJ (2010) Reduced expression of connective tissue growth factor (CTGF/CCN2) mediates collagen loss in chronologically aged human skin. J Investig Dermatol 130:415–424

47 Quan T, Wang F, Shao Y, Rittié L, Xia W, Orringer JS, Voorhees JJ, Fisher GJ (2013) Enhancing structural support of the dermal microenvironment activates fibroblasts, endothelial cells, and keratinocytes in aged human skin in vivo. J Investig Dermatol 133:658–667

48 Rabbany SY, Pastore J, Yamamoto M, Miller T, Rafii S, Aras R, Penn M (2010) Continuous delivery of stromal cell-derived factor-1 from alginate scaffolds accelerateswoundhealing. CellTranspl 19:399–408

49 Ridky TW, Chow JM, Wong DJ, Khavari PA (2010) Invasive threedimensional organotypic neoplasia from multiple normal human epithelia. Nat Med 16:1450–1455

50 Sarchio SN, Scolyer RA, Beaugie C, McDonald D, Marsh-Wakefield F, Halliday GM, Byrne SN (2014) Pharmacologically antagonizing the CXCR4-CXCL12 chemokine pathway with AMD3100 inhibits sunlight-induced skin cancer. J Investig Dermatol 134:1091–1100

51 Sarkar A, Tatlidede S, Scherer SS, Orgill DP, Berthiaume F (2011) Combination of stromal cell-derived factor-1 and collagen-glycosaminoglycan scaffold delays contraction and accelerates reepithelialization of dermal wounds in wild-type mice. Wound Repair Regener 19:71–79

52 Straussman R, Morikawa T, Shee K, Barzily-Rokni M, Qian ZR, Du J, Davis A,Mongare MM, Gould J, Frederick DT, Cooper ZA, Chapman PB, Solit DB,Ribas A, Lo RS, Flaherty KT, Ogino S,Wargo JA,Golub TR(2012)Tumourmicro-environment elicits innate resistance toRAF inhibitors through HGF secretion. Nature 487:500–504

53 Sugiyama T, Kohara H, Noda M, Nagasawa T (2006) Maintenance of the hematopoietic stem cell pool by CXCL12-CXCR4 chemokine signaling in bone marrow stromal cell niches. Immunity 25:977–988

54 Tuan TL, Keller LC, Sun D, Nimni ME, Cheung D (1994) Dermal fibroblasts activate keratinocyte outgrowth on collagen gels. J Cell Sci 107(Pt 8):2285–2289

55 Uitto J (1986) Connective tissue biochemistry of the aging dermis. Agerelated alterations in collagen and elastin. Dermatol Clin 4:433–446

56 Wang J, Wang J, Sun Y, Song W, Nor JE, Wang CY, Taichman RS (2005) Diverse signaling pathways through the SDF-1/CXCR4 chemokine axis in prostate cancer cell lines leads to altered patterns of cytokine secretion and angiogenesis. Cell Signal 17:1578–1592

57 Wright DE, Bowman EP, Wagers AJ, Butcher EC, Weissman IL (2002) Hematopoietic stem cells are uniquely selective in their migratory response to chemokines. J Exp Med 195:1145–1154

58 Zgraggen S, Huggenberger R, Kerl K, Detmar M (2014) An important role of the SDF-1/CXCR4 axis in chronic skin inflammation. PloS One9:e93665