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NRP-1 interacts with GIPC1 and α6/β4-integrins to increase YAP1/∆Np63α-dependent epidermal cancer stem cell survival

Oncogene volume 37, pages 4711–4722 (2018)Cite this article

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Abstract

We have identified an epidermal cancer stem (ECS) cell population that drives formation of rapidly growing and highly invasive and vascularized tumors. VEGF-A and neuropilin-1 (NRP-1) are highly expressed in ECS cell tumors and VEGF-A/NRP-1 interaction is required for ECS cell survival and tumor vascularization. We now identify a novel signaling cascade that is triggered by VEGF-A/NRP-1. We show that NRP-1 forms a complex with GIPC1 and α6/β4-integrin to activate FAK/Src signaling, which leads to stabilization of a YAP1/∆Np63α to enhance ECS cell survival, invasion, and angiogenesis. Loss of NRP-1, GIPC1, α6/β4-integrins, YAP1, or ∆Np63α reduces these responses. Moreover, restoration of constituently active YAP1 or ∆Np63α in NRP-1 null cells restores the ECS cell phenotype. Tumor xenograft experiments show that NRP-1 knockout ECS cells form small tumors characterized by reduced vascularization as compared to wild-type cells. The NRP-1 knockout tumors display signaling changes consistent with a role for the proposed signaling cascade. These studies suggest that VEGF-A interacts with NRP-1 and GIPC1 to regulate α6/β4-integrin, FAK, Src, PI3K/PDK1, LATS1 signaling to increase YAP1/∆Np63α accumulation to drive ECS cell survival, angiogenesis, and tumor formation.

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References

  1. American Cancer Society Website. Cancer facts and figures, http://www cancer org/2010, Accessed April 2017.

  2. Bickers DR, Lim HW, Margolis D, Weinstock MA, Goodman C, Faulkner E. et al. The burden of skin diseases: 2004 a joint project of the American Academy of Dermatology Association and the Society for Investigative Dermatology. J Am Acad Dermatol. 2006;55:490–500.

    Article  PubMed  Google Scholar 

  3. Rogers HW, Weinstock MA, Harris AR, Hinckley MR, Feldman SR, Fleischer AB. et al. Incidence estimate of nonmelanoma skin cancer in the United States, 2006. Arch Dermatol. 2010;146:283–7.

    Article  PubMed  Google Scholar 

  4. Johnson KE, Wilgus TA. Multiple roles for VEGF in non-melanoma skin cancer: angiogenesis and beyond. J Skin Cancer. 2012;2012:483439

    Article  PubMed  PubMed Central  Google Scholar 

  5. Claesson-Welsh L, Welsh M. VEGFA and tumour angiogenesis. J Intern Med. 2013;273:114–27.

    Article  PubMed  CAS  Google Scholar 

  6. Koch S, Tugues S, Li X, Gualandi L, Claesson-Welsh L. Signal transduction by vascular endothelial growth factor receptors. Biochem J. 2011;437:169–83.

    Article  PubMed  CAS  Google Scholar 

  7. Beck B, Driessens G, Goossens S, Youssef KK, Kuchnio A, Caauwe A. et al. A vascular niche and a VEGF-Nrp1 loop regulate the initiation and stemness of skin tumours. Nature. 2011;478:399–403.

    Article  PubMed  CAS  Google Scholar 

  8. Lichtenberger BM, Tan PK, Niederleithner H, Ferrara N, Petzelbauer P, Sibilia M. Autocrine VEGF signaling synergizes with EGFR in tumor cells to promote epithelial cancer development. Cell. 2010;140:268–79.

    Article  PubMed  CAS  Google Scholar 

  9. Man XY, Yang XH, Cai SQ, Yao YG, Zheng M. Immunolocalization and expression of vascular endothelial growth factor receptors (VEGFRs) and neuropilins (NRPs) on keratinocytes in human epidermis. Mol Med. 2006;12:127–36.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Yang XH, Man XY, Cai SQ, Yao YG, Bu ZY, Zheng M. Expression of VEGFR-2 on HaCaT cells is regulated by VEGF and plays an active role in mediating VEGF induced effects. Biochem Biophys Res Commun. 2006;349:31–38.

    Article  PubMed  CAS  Google Scholar 

  11. Zhu JW, Wu XJ, Luo D, Lu ZF, Cai SQ, Zheng M. Activation of VEGFR-2 signaling in response to moderate dose of ultraviolet B promotes survival of normal human keratinocytes. Int J Biochem Cell Biol. 2012;44:246–56.

    Article  PubMed  CAS  Google Scholar 

  12. Hirakawa S, Kodama S, Kunstfeld R, Kajiya K, Brown LF, Detmar M. VEGF-A induces tumor and sentinel lymph node lymphangiogenesis and promotes lymphatic metastasis. J Exp Med. 2005;201:1089–99.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  13. Larcher F, Murillas R, Bolontrade M, Conti CJ, Jorcano JL. VEGF/VPF overexpression in skin of transgenic mice induces angiogenesis, vascular hyperpermeability and accelerated tumor development. Oncogene. 1998;17:303–11.

    Article  PubMed  CAS  Google Scholar 

  14. Bao S, Wu Q, Sathornsumetee S, Hao Y, Li Z, Hjelmeland AB. et al. Stem cell-like glioma cells promote tumor angiogenesis through vascular endothelial growth factor. Cancer Res. 2006;66:7843–8.

    Article  PubMed  CAS  Google Scholar 

  15. Goel HL, Mercurio AM. VEGF targets the tumour cell. Nat Rev Cancer. 2013;13:871–82.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  16. Zhao Y, Bao Q, Renner A, Camaj P, Eichhorn M, Ischenko I. et al. Cancer stem cells and angiogenesis. Int J Dev Biol. 2011;55:477–82.

    Article  PubMed  CAS  Google Scholar 

  17. Casanova ML, Larcher F, Casanova B, Murillas R, Fernandez-Acenero MJ, Villanueva C. et al. A critical role for ras-mediated, epidermal growth factor receptor-dependent angiogenesis in mouse skin carcinogenesis. Cancer Res. 2002;62:3402–7.

    PubMed  CAS  Google Scholar 

  18. Linde N, Lederle W, Depner S, van RN, Gutschalk CM, Mueller MM. Vascular endothelial growth factor-induced skin carcinogenesis depends on recruitment and alternative activation of macrophages. J Pathol. 2012;227:17–28.

    Article  PubMed  CAS  Google Scholar 

  19. Adhikary G, Grun D, Kerr C, Balasubramanian S, Rorke EA, Vemuri M. et al. Identification of a population of epidermal squamous cell carcinoma cells with enhanced potential for tumor formation. PLoS ONE. 2013;8:e84324

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  20. Fisher ML, Kerr C, Adhikary G, Grun D, Xu W, Keillor JW. et al. Transglutaminase interaction with a6/b4-integrin to stimulates YAP1-dependent DNp63a stabilization and leads to enhanced cancer stem cell survival and tumor formation. Cancer Res. 2016;76:7265–76.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  21. Fisher ML, Adhikary G, Xu W, Kerr C, Keillor JW, Eckert RL. Type II transglutaminase stimulates epidermal cancer stem cell epithelial-mesenchymal transition. Oncotarget. 2015;6:20525–39.

    PubMed  PubMed Central  Google Scholar 

  22. Fisher ML, Keillor JW, Xu W, Eckert RL, Kerr C. Transglutaminase is required for epidermal squamous cell carcinoma stem cell survival. Mol Cancer Res. 2015;13:1083–94.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Grun D, Adhikary G, Eckert RL. VEGF-A acts via neuropilin-1 to enhance epidermal cancer stem cell survival and formation of aggressive and highly vascularized tumors. Oncogene. 2016;35:4379–87.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  24. Yoshida A, Shimizu A, Asano H, Kadonosono T, Kondoh SK, Geretti E. et al. VEGF-A/NRP1 stimulates GIPC1 and Syx complex formation to promote RhoA activation and proliferation in skin cancer cells. Biol Open. 2015;4:1063–76.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Harvey KF, Zhang X, Thomas DM. The Hippo pathway and human cancer. Nat Rev Cancer. 2013;13:246–57.

    Article  PubMed  CAS  Google Scholar 

  26. Lampropoulou A, Ruhrberg C. Neuropilin regulation of angiogenesis. Biochem Soc Trans. 2014;42:1623–8.

    Article  PubMed  CAS  Google Scholar 

  27. Katoh M. Functional proteomics, human genetics and cancer biology of GIPC family members. Exp Mol Med. 2013;45:e26

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  28. Valdembri D, Caswell PT, Anderson KI, Schwarz JP, Konig I, Astanina E. et al. Neuropilin-1/GIPC1 signaling regulates alpha5beta1 integrin traffic and function in endothelial cells. PLoS Biol. 2009;7:e25

    Article  PubMed  CAS  Google Scholar 

  29. Zhang G, Chen L, Sun K, Khan AA, Yan J, Liu H. et al. Neuropilin-1 (NRP-1)/GIPC1 pathway mediates glioma progression. Tumour Biol. 2016;37:13777–88.

    Article  PubMed  CAS  Google Scholar 

  30. El MH, Poinat P, Koster J, Sondermann H, Wixler V, Wegener E. et al. The PDZ domain of TIP-2/GIPC interacts with the C-terminus of the integrin alpha5 and alpha6 subunits. Matrix Biol. 2002;21:207–14.

    Article  Google Scholar 

  31. Tani TT, Mercurio AM. PDZ interaction sites in integrin alpha subunits. T14853, TIP/GIPC binds to a type I recognition sequence in alpha 6A/alpha 5 and a novel sequence in alpha 6B. J Biol Chem. 2001;276:36535–42.

    Article  PubMed  CAS  Google Scholar 

  32. Boukamp P, Petrussevska RT, Breitkreutz D, Hornung J, Markham A, Fusenig NE. Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell Biol. 1988;106:761–71.

    Article  PubMed  CAS  Google Scholar 

  33. Adhikary G, Grun D, Balasubramanian S, Kerr C, Huang J, Eckert R. Survival of skin cancer stem cells requires the Ezh2 polycomb group protein. Carcinogenesis. 2015;36:800–10.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  34. Djordjevic S, Driscoll PC. Targeting VEGF signalling via the neuropilin co-receptor. Drug Discov Today. 2013;18:447–55.

    Article  PubMed  CAS  Google Scholar 

  35. Mamluk R, Gechtman Z, Kutcher ME, Gasiunas N, Gallagher J, Klagsbrun M. Neuropilin-1 binds vascular endothelial growth factor 165, placenta growth factor-2, and heparin via its b1b2 domain. J Biol Chem. 2002;277:24818–25.

    Article  PubMed  CAS  Google Scholar 

  36. Migdal M, Huppertz B, Tessler S, Comforti A, Shibuya M, Reich R. et al. Neuropilin-1 is a placenta growth factor-2 receptor. J Biol Chem. 1998;273:22272–8.

    Article  PubMed  CAS  Google Scholar 

  37. Soker S, Takashima S, Miao HQ, Neufeld G, Klagsbrun M. Neuropilin-1 is expressed by endothelial and tumor cells as an isoform-specific receptor for vascular endothelial growth factor. Cell. 1998;92:735–45.

    Article  PubMed  CAS  Google Scholar 

  38. Gray MJ, Wey JS, Belcheva A, McCarty MF, Trevino JG, Evans DB. et al. Neuropilin-1 suppresses tumorigenic properties in a human pancreatic adenocarcinoma cell line lacking neuropilin-1 coreceptors. Cancer Res. 2005;65:3664–70.

    Article  PubMed  CAS  Google Scholar 

  39. Jarvis A, Allerston CK, Jia H, Herzog B, Garza-Garcia A, Winfield N. et al. Small molecule inhibitors of the neuropilin-1 vascular endothelial growth factor A (VEGF-A) interaction. J Med Chem. 2010;53:2215–26.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  40. Jia H, Cheng L, Tickner M, Bagherzadeh A, Selwood D, Zachary I. Neuropilin-1 antagonism in human carcinoma cells inhibits migration and enhances chemosensitivity. Br J Cancer. 2010;102:541–52.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  41. Li M, Yang H, Chai H, Fisher WE, Wang X, Brunicardi FC. et al. Pancreatic carcinoma cells express neuropilins and vascular endothelial growth factor, but not vascular endothelial growth factor receptors. Cancer. 2004;101:2341–50.

    Article  PubMed  CAS  Google Scholar 

  42. Cao Y, E G, Wang E, Pal K, Dutta SK, Bar-Sagi D. et al. VEGF exerts an angiogenesis-independent function in cancer cells to promote their malignant progression. Cancer Res. 2012;72:3912–8.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  43. Kim J, Kim YH, Kim J, Park DY, Bae H, Lee DH. et al. YAP/TAZ regulates sprouting angiogenesis and vascular barrier maturation. J Clin Invest. 2017;127:3441–61.

    Article  PubMed  PubMed Central  Google Scholar 

  44. Wang X, Freire VA, Schermann G, Shen Y, Moya IM, Castro L. et al. YAP/TAZ orchestrate VEGF signaling during developmental angiogenesis. Dev Cell. 2017;42:462–78.

    Article  PubMed  CAS  Google Scholar 

  45. Elbediwy A, Vincent-Mistiaen ZI, Spencer-Dene B, Stone RK, Boeing S, Wculek SK. et al. Integrin signalling regulates YAP and TAZ to control skin homeostasis. Development. 2016;143:1674–87.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  46. Koster MI, Roop DR. The role of p63 in development and differentiation of the epidermis. J Dermatol Sci. 2004;34:3–9.

    Article  PubMed  CAS  Google Scholar 

  47. Bowden J, Brennan PA, Umar T, Cronin A. Expression of vascular endothelial growth factor in basal cell carcinoma and cutaneous squamous cell carcinoma of the head and neck. J Cutan Pathol. 2002;29:585–9.

    Article  PubMed  Google Scholar 

  48. Shahrabi-Farahani S, Gallottini M, Martins F, Li E, Mudge DR, Nakayama H. et al. Neuropilin 1 receptor is up-regulated in dysplastic epithelium and oral squamous cell carcinoma. Am J Pathol. 2016;186:1055–64.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  49. Siegle JM, Basin A, Sastre-Perona A, Yonekubo Y, Brown J, Sennett R. et al. SOX2 is a cancer-specific regulator of tumour initiating potential in cutaneous squamous cell carcinoma. Nat Commun. 2014;5:4511

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  50. Sniezek JC, Matheny KE, Westfall MD, Pietenpol JA. Dominant negative p63 isoform expression in head and neck squamous cell carcinoma. Laryngoscope. 2004;114:2063–72.

    Article  PubMed  CAS  Google Scholar 

  51. Thurfjell N, Coates PJ, Uusitalo T, Mahani D, Dabelsteen E, Dahlqvist A. et al. Complex p63 mRNA isoform expression patterns in squamous cell carcinoma of the head and neck. Int J Oncol. 2004;25:27–35.

    PubMed  CAS  Google Scholar 

  52. Lamar JM, Stern P, Liu H, Schindler JW, Jiang ZG, Hynes RO. The Hippo pathway target, YAP, promotes metastasis through its TEAD-interaction domain. Proc Natl Acad Sci USA. 2012;109:E2441–50.

    Article  PubMed  Google Scholar 

  53. Rheinwald JG, Beckett MA. Tumorigenic keratinocyte lines requiring anchorage and fibroblast support cultures from human squamous cell carcinomas. Cancer Res. 1981;41:1657–63.

    PubMed  CAS  Google Scholar 

  54. Chew YC, Adhikary G, Xu W, Wilson GM, Eckert RL. Protein kinase C delta increases Kruppel-like factor 4 protein, which drives involucrin gene transcription in differentiating keratinocytes. J Biol Chem. 2013;288:17759–68.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  55. Fisher ML, Adhikary G, Grun D, Kaetzel DM, Eckert RL. The Ezh2 polycomb group protein drives an aggressive phenotype in melanoma cancer stem cells and is a target of diet derived sulforaphane. Mol Carcinog. 2015;55:2024–36.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

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Acknowledgements

This work was supported by the NIH (CA131074 and CA184027) to RLE and a pilot grant from the Greenebaum Comprehensive Cancer Center (P30 CA134274).

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    Daniel Grun, Gautam Adhikary & Richard L. Eckert

  2. Department of Dermatology, University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    Richard L. Eckert

  3. Department of Reproductive Biology, University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    Richard L. Eckert

  4. Marlene and Stewart Greenebaum Comprehensive Cancer Center, University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    Richard L. Eckert

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Grun, D., Adhikary, G. & Eckert, R.L. NRP-1 interacts with GIPC1 and α6/β4-integrins to increase YAP1/∆Np63α-dependent epidermal cancer stem cell survival. Oncogene 37, 4711–4722 (2018). https://doi.org/10.1038/s41388-018-0290-4

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