Extracellular vesicle-dependent effect of RNA-binding protein IGF2BP1 on melanoma metastasis


Insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) is a multifunctional RNA-binding protein with an oncofetal pattern of expression shown to be implicated in the development of a variety of malignancies. In this study, we explored the role and mechanisms of IGF2BP1 in melanoma development and progression. In two different in vivo models, we showed that although genetic deletion or shRNA-mediated suppression of IGF2BP1 did not affect primary tumor formation, it drastically suppressed lung metastasis. Here we demonstrated that extracellular vesicles (EVs) secreted by melanoma cells mediate the effects of IGF2BP1 on metastasis: EVs from the IGF2BP1 knockdown melanoma cells failed to promote metastasis, whereas EVs isolated from IGF2BP1-overexpressed melanoma cells further accelerated EV-induced metastasis. Moreover, the EVs from IGF2BP1 knockdown melanoma cells inhibited fibronectin deposition and accumulation of CD45+ cells in the lungs compared with control EVs, thus blocking the pre-metastatic niche formation potential of EVs. IGF2BP1 knockdown did not affect size, number, or protein/RNA concentration of secreted EVs or their uptake by recipient cells in vitro or in vivo. However, RNA-sequencing and proteomics analysis of the EVs revealed differential expression in a number of mRNA, proteins, and miRNAs. This suggested that IGF2BP1 is intimately involved in the regulation of the cargo of EVs, thereby affecting the pro-metastatic function of melanoma-derived EVs. To the best of our knowledge, this is the first study that demonstrates the role of RNA-binding protein IGF2BP1 in EV-mediated promotion of melanoma metastasis and may provide novel avenues for the development of metastatic inhibitors.

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  1. 1.

    Dimitriadis E, Trangas T, Milatos S, Foukas PG, Gioulbasanis I, Courtis N, et al. Expression of oncofetal RNA-binding protein CRD-BP/IMP1 predicts clinical outcome in colon cancer. Int J Cancer. 2007;121:486–94.

    CAS  Article  Google Scholar 

  2. 2.

    Bell JL, Turlapati R, Liu T, Schulte JH, Huttelmaier S. IGF2BP1 harbors prognostic significance by gene gain and diverse expression in neuroblastoma. J Clin Oncol. 2015;33:1285–93.

    CAS  Article  Google Scholar 

  3. 3.

    Vikesaa J, Hansen TV, Jonson L, Borup R, Wewer UM, Christiansen J, et al. RNA-binding IMPs promote cell adhesion and invadopodia formation. EMBO J. 2006;25:1456–68.

    CAS  Article  Google Scholar 

  4. 4.

    Elcheva I, Goswami S, Noubissi FK, Spiegelman VS. CRD-BP protects the coding region of betaTrCP1 mRNA from miR-183-mediated degradation. Mol Cell. 2009;35:240–6.

    CAS  Article  Google Scholar 

  5. 5.

    Goswami S, Tarapore RS, Poenitzsch Strong AM, TeSlaa JJ, Grinblat Y, Setaluri V, et al. MicroRNA-340-mediated degradation of microphthalmia-associated transcription factor (MITF) mRNA is inhibited by coding region determinant-binding protein (CRD-BP). J Biol Chem. 2015;290:384–95.

    CAS  Article  Google Scholar 

  6. 6.

    Noubissi FK, Elcheva I, Bhatia N, Shakoori A, Ougolkov A, Liu J, et al. CRD-BP mediates stabilization of betaTrCP1 and c-myc mRNA in response to beta-catenin signalling. Nature. 2006;441:898–901.

    CAS  Article  Google Scholar 

  7. 7.

    Noubissi FK, Goswami S, Sanek NA, Kawakami K, Minamoto T, Moser A, et al. Wnt signaling stimulates transcriptional outcome of the Hedgehog pathway by stabilizing GLI1 mRNA. Cancer Res. 2009;69:8572–8.

    CAS  Article  Google Scholar 

  8. 8.

    Sparanese D, Lee CH. CRD-BP shields c-myc and MDR-1 RNA from endonucleolytic attack by a mammalian endoribonuclease. Nucleic Acids Res. 2007;35:1209–21.

    CAS  Article  Google Scholar 

  9. 9.

    Leeds P, Kren BT, Boylan JM, Betz NA, Steer CJ, Gruppuso PA, et al. Developmental regulation of CRD-BP, an RNA-binding protein that stabilizes c-myc mRNA in vitro. Oncogene. 1997;14:1279–86.

    CAS  Article  Google Scholar 

  10. 10.

    Stohr N, Huttelmaier S. IGF2BP1: a post-transcriptional “driver” of tumor cell migration. Cell Adh Migr. 2012;6:312–8.

    Article  Google Scholar 

  11. 11.

    Elcheva I, Tarapore RS, Bhatia N, Spiegelman VS. Overexpression of mRNA-binding protein CRD-BP in malignant melanomas. Oncogene. 2008;27:5069–74.

    CAS  Article  Google Scholar 

  12. 12.

    Craig EA, Spiegelman VS. Inhibition of CRD-BP sensitizes melanoma cells to chemotherapeutic agents. Pigment Cell Melanoma Res. 2012;25:83–7.

    CAS  Article  Google Scholar 

  13. 13.

    Kim T, Havighurst T, Kim K, Albertini M, Xu YG, Spiegelman VS. Targeting insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) in metastatic melanoma to increase efficacy of BRAF(V600E) inhibitors. Mol Carcinog. 2018;57:678–83.

    CAS  Article  Google Scholar 

  14. 14.

    Peinado H, Alečković M, Lavotshkin S, Matei I, Costa-Silva B, Moreno-Bueno G, et al. Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET. Nat Med. 2012;18:883.

    CAS  Article  Google Scholar 

  15. 15.

    Azmi AS, Bao B, Sarkar FH. Exosomes in cancer development, metastasis, and drug resistance: a comprehensive review. Cancer Metastas- Rev. 2013;32:623–42.

    CAS  Article  Google Scholar 

  16. 16.

    Chen Y, Zeng C, Zhan Y, Wang H, Jiang X, Li W. Aberrant low expression of p85alpha in stromal fibroblasts promotes breast cancer cell metastasis through exosome-mediated paracrine Wnt10b. Oncogene. 2017;36:4692–705.

    CAS  Article  Google Scholar 

  17. 17.

    Weidle HU, Birzele F, Kollmorgen G, RÜGer R. The multiple roles of exosomes in metastasis. Cancer Genom Proteom. 2017;14:1–16.

    CAS  Article  Google Scholar 

  18. 18.

    Lobb RJ, Lima LG, Moller A. Exosomes: key mediators of metastasis and pre-metastatic niche formation. Semin Cell Dev Biol. 2017;67:3–10.

    CAS  Article  Google Scholar 

  19. 19.

    Hood JL, San RS, Wickline SA. Exosomes released by melanoma cells prepare sentinel lymph nodes for tumor metastasis. Cancer Res. 2011;71:3792–801.

    CAS  Article  Google Scholar 

  20. 20.

    Costa-Silva B, Aiello NM, Ocean AJ, Singh S, Zhang H, Thakur Basant K, et al. Pancreatic cancer exosomes initiate pre-metastatic niche formation in the liver. Nat Cell Biol. 2015;17:816.

    CAS  Article  Google Scholar 

  21. 21.

    Dankort D, Curley DP, Cartlidge RA, Nelson B, Karnezis AN, Damsky WE Jr., et al. Braf(V600E) cooperates with Pten loss to induce metastatic melanoma. Nat Genet. 2009;41:544–52.

    CAS  Article  Google Scholar 

  22. 22.

    Hamilton KE, Noubissi FK, Katti PS, Hahn CM, Davey SR, Lundsmith ET, et al. IMP1 promotes tumor growth, dissemination and a tumor-initiating cell phenotype in colorectal cancer cell xenografts. Carcinogenesis. 2013;34:2647–54.

    CAS  Article  Google Scholar 

  23. 23.

    Wen SW, Sceneay J, Lima LG, Wong CS, Becker M, Krumeich S, et al. The biodistribution and immune suppressive effects of breast cancer-derived exosomes. Cancer Res. 2016;76:6816–27.

    CAS  Article  Google Scholar 

  24. 24.

    TaeWon K, Thomas H, KyungMann K, Mark A, G XY, S SV. Targeting insulin‐like growth factor 2 mRNA‐binding protein 1 (IGF2BP1) in metastatic melanoma to increase efficacy of BRAFV600E inhibitors. Mol Carcinog. 2018;57:678–83.

    Article  Google Scholar 

  25. 25.

    Wang R-j, Li J-w, Bao B-h, Wu H-c, Du Z-h, Su J-l, et al. MicroRNA-873 (miRNA-873) inhibits glioblastoma tumorigenesis and metastasis by suppressing the expression of IGF2BP1. J Biol Chem. 2015;290:8938–48.

    CAS  Article  Google Scholar 

  26. 26.

    Su Y, Xiong J, Hu J, Wei X, Zhang X, Rao L. MicroRNA-140-5p targets insulin like growth factor 2 mRNA binding protein 1 (IGF2BP1) to suppress cervical cancer growth and metastasis. Oncotarget. 2016;7:68397–411.

    PubMed  PubMed Central  Google Scholar 

  27. 27.

    Hoshino A, Costa-Silva B, Shen T-L, Rodrigues G, Hashimoto A, Tesic Mark M, et al. Tumour exosome integrins determine organotropic metastasis. Nature. 2015;527:329.

    CAS  Article  Google Scholar 

  28. 28.

    Liu Y, Gu Y, Han Y, Zhang Q, Jiang Z, Zhang X, et al. Tumor exosomal RNAs promote lung pre-metastatic niche formation by activating alveolar epithelial TLR3 to recruit neutrophils. Cancer Cell. 2016;30:243–56.

    Article  Google Scholar 

  29. 29.

    Peinado H, Lavotshkin S, Lyden D. The secreted factors responsible for pre-metastatic niche formation: old sayings and new thoughts. Semin Cancer Biol. 2011;21:139–46.

    CAS  Article  Google Scholar 

  30. 30.

    Psaila B, Lyden D. The metastatic niche: adapting the foreign soil. Nat Rev Cancer. 2009;9:285–93.

    CAS  Article  Google Scholar 

  31. 31.

    Li W, Hu Y, Jiang T, Han Y, Han G, Chen J, et al. Rab27A regulates exosome secretion from lung adenocarcinoma cells A549: involvement of EPI64. APMIS. 2014;122:1080–7.

    CAS  PubMed  Google Scholar 

  32. 32.

    Dorayappan KDP, Wanner R, Wallbillich JJ, Saini U, Zingarelli R, Suarez AA, et al. Hypoxia-induced exosomes contribute to a more aggressive and chemoresistant ovarian cancer phenotype: a novel mechanism linking STAT3/Rab proteins. Oncogene. 2018;37:3806–21.

    CAS  Article  Google Scholar 

  33. 33.

    Meerbrey KL, Hu G, Kessler JD, Roarty K, Li MZ, Fang JE, et al. The pINDUCER lentiviral toolkit for inducible RNA interference in vitro and in vivo. Proc Natl Acad Sci USA. 2011;108:3665–70.

    CAS  Article  Google Scholar 

  34. 34.

    Ghoshal A, Ghosh SS. Antagonizing canonical Wnt signaling pathway by recombinant human sFRP4 purified from E. coli and its implications in cancer therapy. Mol Cell Biochem. 2016;418:119–35.

    CAS  Article  Google Scholar 

  35. 35.

    Iyer SC, Gopal A, Halagowder D. Myricetin induces apoptosis by inhibiting P21 activated kinase 1 (PAK1) signaling cascade in hepatocellular carcinoma. Mol Cell Biochem. 2015;407:223–37.

    CAS  Article  Google Scholar 

  36. 36.

    Bosenberg M, Muthusamy V, Curley DP, Wang Z, Hobbs C, Nelson B, et al. Characterization of melanocyte-specific inducible Cre recombinase transgenic mice. Genesis. 2006;44:262–7.

    CAS  Article  Google Scholar 

  37. 37.

    Oberman F, Rand K, Maizels Y, Rubinstein AM, Yisraeli JK. VICKZ proteins mediate cell migration via their RNA binding activity. RNA (New Y, NY). 2007;13:1558–69.

    CAS  Article  Google Scholar 

  38. 38.

    Gupta S, Rawat S, Arora V, Kottarath SK, Dinda AK, Vaishnav PK, et al. An improvised one-step sucrose cushion ultracentrifugation method for exosome isolation from culture supernatants of mesenchymal stem cells. Stem Cell Res Ther. 2018;9:180.

    Article  Google Scholar 

  39. 39.

    Jao CY, Salic A. Exploring RNA transcription and turnover in vivo by using click chemistry. Proc Natl Acad Sci USA. 2008;105:15779–84.

    CAS  Article  Google Scholar 

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This study was supported in part by the NIH grant R01 AR063361 (VSS). We thank Dr Ze’ev Ronai for the generous gift of reagents and Dr JM Sundstrom for the help with NanosightTM analysis of EVs. We thank Penn State Cancer Institute Genomics Sciences, Penn State College of Medicine Imaging Core, Flow Cytometry Core, and Molecular and Histopathology Core Facilities for help with respective data acquisition and analysis.

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Correspondence to Vladimir S. Spiegelman.

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Ghoshal, A., Rodrigues, L.C., Gowda, C.P. et al. Extracellular vesicle-dependent effect of RNA-binding protein IGF2BP1 on melanoma metastasis. Oncogene 38, 4182–4196 (2019). https://doi.org/10.1038/s41388-019-0797-3

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