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GPR55 signalling promotes proliferation of pancreatic cancer cells and tumour growth in mice, and its inhibition increases effects of gemcitabine

Oncogene volume 37, pages 6368–6382 (2018)Cite this article

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Abstract

The life expectancy for pancreatic cancer patients has seen no substantial changes in the last 40 years as very few and mostly just palliative treatments are available. As the five years survival rate remains around 5%, the identification of novel pharmacological targets and development of new therapeutic strategies are urgently needed. Here we demonstrate that inhibition of the G protein-coupled receptor GPR55, using genetic and pharmacological approaches, reduces pancreatic cancer cell growth in vitro and in vivo and we propose that this may represent a novel strategy to inhibit pancreatic ductal adenocarcinoma (PDAC) progression. Specifically, we show that genetic ablation of Gpr55 in the KRASWT/G12D/TP53WT/R172H/Pdx1-Cre+/+ (KPC) mouse model of PDAC significantly prolonged survival. Importantly, KPC mice treated with a combination of the GPR55 antagonist Cannabidiol (CBD) and gemcitabine (GEM, one of the most used drugs to treat PDAC), survived nearly three times longer compared to mice treated with vehicle or GEM alone. Mechanistically, knockdown or pharmacologic inhibition of GPR55 reduced anchorage-dependent and independent growth, cell cycle progression, activation of mitogen-activated protein kinase (MAPK) signalling and protein levels of ribonucleotide reductases in PDAC cells. Consistent with this, genetic ablation of Gpr55 reduced proliferation of tumour cells, MAPK signalling and ribonucleotide reductase M1 levels in KPC mice. Combination of CBD and GEM inhibited tumour cell proliferation in KPC mice and it opposed mechanisms involved in development of resistance to GEM in vitro and in vivo. Finally, we demonstrate that the tumour suppressor p53 regulates GPR55 protein expression through modulation of the microRNA miR34b-3p. Our results demonstrate the important role played by GPR55 downstream of p53 in PDAC progression. Moreover our data indicate that combination of CBD and GEM, both currently approved for medical use, might be tested in clinical trials as a novel promising treatment to improve PDAC patients’ outcome.

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References

  1. Hruban RH, Goggins M, Parsons J, Kern SE. Progression model for pancreatic cancer. Clin Cancer Res. 2000;6:2969–72.

    CAS  PubMed  Google Scholar 

  2. Hruban RH, Wilentz RE, Kern SE. Genetic progression in the pancreatic ducts. Am J Pathol. 2000;156:1821–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Eser S, Schnieke A, Schneider G, Saur D. Oncogenic KRAS signalling in pancreatic cancer. Br J Cancer. 2014;111:817–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Scarpa A, Capelli P, Mukai K, Zamboni G, Oda T, Iacono C, et al. Pancreatic adenocarcinomas frequently show p53 gene mutations. Am J Pathol. 1993;142:1534–43.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Hingorani SR, Petricoin EF, Maitra A, Rajapakse V, King C, Jacobetz MA, et al. Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse. Cancer Cell. 2003;4:437–50.

    Article  CAS  PubMed  Google Scholar 

  6. Hingorani SR, Wang L, Multani AS, Combs C, Deramaudt TB, Hruban RH, et al. Trp53R172H and KrasG12D cooperate to promote chromosomal instability and widely metastatic pancreatic ductal adenocarcinoma in mice. Cancer Cell. 2005;7:469–83.

    Article  CAS  PubMed  Google Scholar 

  7. Morris JP, Wang SC, Hebrok M. KRAS, Hedgehog, Wnt and the twisted developmental biology of pancreatic ductal adenocarcinoma. Nat Rev Cancer. 2010;10:683–95.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Peixoto RD, Ho M, Renouf DJ, Lim HJ, Gill S, Ruan JY, et al. Eligibility of metastatic pancreatic cancer patients for first-line palliative intent nab-paclitaxel plus gemcitabine versus FOLFIRINOX. Am J Clin Oncol. 2017;40:507–11.

    Article  CAS  PubMed  Google Scholar 

  9. Vaccaro V, Sperduti I, Milella M. FOLFIRINOX versus gemcitabine for metastatic pancreatic cancer. N Engl J Med. 2011;365:768–9. Author reply 769.

    Article  CAS  PubMed  Google Scholar 

  10. Vincent A, Herman J, Schulick R, Hruban RH, Goggins M. Pancreatic cancer. Lancet. 2011;378:607–20.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Falasca M, Kim M, Casari I. Pancreatic cancer: current research and future directions. Biochim Biophys Acta. 2016;1865:123–32.

    CAS  PubMed  Google Scholar 

  12. Oka S, Nakajima K, Yamashita A, Kishimoto S, Sugiura T. Identification of GPR55 as a lysophosphatidylinositol receptor. Biochem Biophys Res Commun. 2007;362:928–34.

    Article  CAS  PubMed  Google Scholar 

  13. Romero-Zerbo SY, Rafacho A, Diaz-Arteaga A, Suarez J, Quesada I, Imbernon M, et al. A role for the putative cannabinoid receptor GPR55 in the islets of Langerhans. J Endocrinol. 2011;211:177–85.

    Article  CAS  PubMed  Google Scholar 

  14. Sisay S, Pryce G, Jackson SJ, Tanner C, Ross RA, Michael GJ, et al. Genetic background can result in a marked or minimal effect of gene knockout (GPR55 and CB2 receptor) in experimental autoimmune encephalomyelitis models of multiple sclerosis. PLoS ONE. 2013;8:e76907.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Roux PP, Shahbazian D, Vu H, Holz MK, Cohen MS, Taunton J, et al. RAS/ERK signaling promotes site-specific ribosomal protein S6 phosphorylation via RSK and stimulates cap-dependent translation. J Biol Chem. 2007;282:14056–64.

    Article  CAS  PubMed  Google Scholar 

  16. He L, He X, Lim LP, de Stanchina E, Xuan Z, Liang Y, et al. A microRNA component of the p53 tumour suppressor network. Nature. 2007;447:1130–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Vogt M, Munding J, Gruner M, Liffers ST, Verdoodt B, Hauk J, et al. Frequent concomitant inactivation of miR-34a and miR-34b/c by CpG methylation in colorectal, pancreatic, mammary, ovarian, urothelial, and renal cell carcinomas and soft tissue sarcomas. Virchows Arch. 2011;458:313–22.

    Article  PubMed  Google Scholar 

  18. Chou TC. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev. 2006;58:621–81.

    Article  CAS  PubMed  Google Scholar 

  19. Zheng C, Jiao X, Jiang Y, Sun S. ERK1/2 activity contributes to gemcitabine resistance in pancreatic cancer cells. J Int Med Res. 2013;41:300–6.

    Article  CAS  PubMed  Google Scholar 

  20. Davidson JD, Ma L, Flagella M, Geeganage S, Gelbert LM, Slapak CA. An increase in the expression of ribonucleotide reductase large subunit 1 is associated with gemcitabine resistance in non-small cell lung cancer cell lines. Cancer Res. 2004;64:3761–6.

    Article  CAS  PubMed  Google Scholar 

  21. Bergman AM, Eijk PP, Ruiz van Haperen VW, Smid K, Veerman G, Hubeek I, et al. In vivo induction of resistance to gemcitabine results in increased expression of ribonucleotide reductase subunit M1 as the major determinant. Cancer Res. 2005;65:9510–6.

    Article  CAS  PubMed  Google Scholar 

  22. Pineiro R, Maffucci T, Falasca M. The putative cannabinoid receptor GPR55 defines a novel autocrine loop in cancer cell proliferation. Oncogene. 2011;30:142–52.

    Article  CAS  PubMed  Google Scholar 

  23. Ross RA. L-α-Lysophosphatidylinositol meets GPR55: a deadly relationship. Trends Pharmacol Sci. 2011;32:265–9.

    Article  CAS  PubMed  Google Scholar 

  24. Falasca M, Corda D. Elevated levels and mitogenic activity of lysophosphatidylinositol in k-ras-transformed epithelial cells. Eur J Biochem. 1994;221:383–9.

    Article  CAS  PubMed  Google Scholar 

  25. Falasca M, Iurisci C, Carvelli A, Sacchetti A, Corda D. Release of the mitogen lysophosphatidylinositol from H-Ras-transformed fibroblasts; a possible mechanism of autocrine control of cell proliferation. Oncogene. 1998;16:2357–65.

    Article  CAS  PubMed  Google Scholar 

  26. Falasca M, Silletta MG, Carvelli A, Di Francesco AL, Fusco A, Ramakrishna V, et al. Signalling pathways involved in the mitogenic action of lysophosphatidylinositol. Oncogene. 1995;10:2113–24.

    CAS  PubMed  Google Scholar 

  27. Ford LA, Roelofs AJ, Anavi-Goffer S, Mowat L, Simpson DG, Irving AJ, et al. A role for L-alpha-lysophosphatidylinositol and GPR55 in the modulation of migration, orientation and polarization of human breast cancer cells. Br J Pharmacol. 2010;160:762–71.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Hofmann NA, Yang J, Trauger SA, Nakayama H, Huang L, Strunk D, et al. The GPR 55 agonist, L-alpha-lysophosphatidylinositol, mediates ovarian carcinoma cell-induced angiogenesis. Br J Pharmacol. 2015;172:4107–18.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Sutphen R, Xu Y, Wilbanks GD, Fiorica J, Grendys EC Jr., LaPolla JP, et al. Lysophospholipids are potential biomarkers of ovarian cancer. Cancer Epidemiol Biomark Prev. 2004;13:1185–91.

    CAS  Google Scholar 

  30. Kargl J, Andersen L, Hasenohrl C, Feuersinger D, Stancic A, Fauland A, et al. GPR55 promotes migration and adhesion of colon cancer cells indicating a role in metastasis. Br J Pharmacol. 2016;173:142–54.

    Article  CAS  PubMed  Google Scholar 

  31. Andradas C, Caffarel MM, Perez-Gomez E, Salazar M, Lorente M, Velasco G, et al. The orphan G protein-coupled receptor GPR55 promotes cancer cell proliferation via ERK. Oncogene. 2011;30:245–52.

    Article  CAS  PubMed  Google Scholar 

  32. Perez-Gomez E, Andradas C, Flores JM, Quintanilla M, Paramio JM, Guzman M, et al. The orphan receptor GPR55 drives skin carcinogenesis and is upregulated in human squamous cell carcinomas. Oncogene. 2013;32:2534–42.

    Article  CAS  PubMed  Google Scholar 

  33. Andradas C, Blasco-Benito S, Castillo-Lluva S, Dillenburg-Pilla P, Diez-Alarcia R, Juanes-Garcia A, et al. Activation of the orphan receptor GPR55 by lysophosphatidylinositol promotes metastasis in triple-negative breast cancer. Oncotarget. 2016;7:47565–75.

    Article  PubMed  PubMed Central  Google Scholar 

  34. Paul RK, Wnorowski A, Gonzalez-Mariscal I, Nayak SK, Pajak K, Moaddel R, et al. (R,R ‘)-4 ‘-methoxy-1-naphthylfenoterol targets GPR55-mediated ligand internalization and impairs cancer cell motility. Biochem. Pharmacol. 2014;87:547–61.

    Article  CAS  PubMed  Google Scholar 

  35. Ji Q, Hao XB, Zhang M,Tang WH, Meng Y, Li L, et al. MicroRNA miR-34 inhibits human pancreatic cancer tumor-initiating cells. PLoS ONE 2009;4:e6816.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Suresh K. An overview of randomization techniques: An unbiased assessment of outcome in clinical research. J Hum Reprod Sci. 2011;4:8–11.

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by Pancreatic Cancer Research Fund and Avner Pancreatic Cancer Foundation (grants to MF). RF, CAF, CEE were supported by Pancreatic Cancer Research Fund (grants to MF). AA is supported by Curtin International Postgraduate Research Scholarship (CIPRS)/Health Sciences Faculty International Research Scholarship (HSFIRS). GS was supported by Ministero Sanità Finalizzata 2011/2012. VDL was supported by AIRC IG 15196.c. We thank Prof David A.Tuveson and Prof David Baker for KC/KPC and GPR55−/− mice respectively; Dr Massimo Broggini for p53 mutants constructs; Prof Hemant Kocher for HPDE cells; GW Pharmaceuticals for providing the cannabidiol used in this study; the Pathology Core facility of the Blizard Institute for helping with IHC. AA, VDL and MF also acknowledge the infrastructure and staff support provided by CHIRI, School of Pharmacy & Biomedical Sciences and by Faculty of Health Sciences, Curtin University.

Author contributions

RF and MF designed, coordinated, and carried out the bulk of the experiments. RF, AA, IM, TM and MF performed in vitro experiments. RF, TM, and MF designed and supervised in vitro experiments. RF, GS, and MF designed and supervised in vivo experiments. RF performed in vivo experiments. SAA, CAF, CEE, and VDL contributed to in vivo experiments. RF and RL performed IHC assay. RL and MP performed IHC analysis. LS and GC performed in silico analysis. OJS provided key reagents. RF, TM, and MF wrote the manuscript. MF conceived the project, led and supervised the study.

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Authors and Affiliations

  1. Queen Mary University of London, Barts and The London School of Medicine and Dentistry, Blizard Institute, Centre for Cell Biology and Cutaneous Research, 4 Newark Street, London, E1 2AT, UK

    R. Ferro, I. Mavrommati, C. E. Edling, S. A. Arifin, C. A. Fyffe, T. Maffucci & M. Falasca

  2. Metabolic Signalling Group, School of Pharmacy & Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, 6102, Perth, WA, Australia

    A. Adamska, V. De Laurenzi & M. Falasca

  3. Dipartimento di Scienze Mediche, Orali e Biotecnologiche, University “G. d’Annunzio” di Chieti-Pescara, Centro Studi sull’Invecchiamento, CeSI-MeT, Chieti, 66100, Italy

    R. Lattanzio, G. Sala, V. De Laurenzi & M. Piantelli

  4. Cancer Genomics Laboratory, Fondazione Edo and Elvo Tempia, Biella, Italy

    L. Sacchetto & G. Chiorino

  5. Cancer Research UK Beatson Institute, Glasgow, G61 1BD, UK

    O. J. Sansom

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  1. R. Ferro
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Correspondence to M. Falasca.

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Ferro, R., Adamska, A., Lattanzio, R. et al. GPR55 signalling promotes proliferation of pancreatic cancer cells and tumour growth in mice, and its inhibition increases effects of gemcitabine. Oncogene 37, 6368–6382 (2018). https://doi.org/10.1038/s41388-018-0390-1

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