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AKT as a key target for growth promoting functions of neutral ceramidase in colon cancer cells

Oncogene volume 37, pages 3852–3863 (2018)Cite this article

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Abstract

Despite advances in the field, colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide. Research into bioactive sphingolipids over the past two decades has played an important role in increasing our understanding of the pathogenesis and therapeutics of CRC. In the complex metabolic network of sphingolipids, ceramidases (CDases) have a key function. These enzymes hydrolyze ceramides into sphingosine (SPH) which in turn is phosphorylated by sphingosine kinases (SK) 1 and 2 to generate sphingosine-1 phosphate (S1P). Importantly, we have recently shown that inhibition of neutral CDase (nCDase) induces an increase of ceramide in colon cancer cells which decreases cellular growth, increases apoptosis and modulates the WNT/β-catenin pathway. We have also shown that the deletion of nCDase protected mice from the onset and progression of colorectal cancer in the AOM carcinogen model. Here, we demonstrate that AKT is a key target for the growth suppressing functions of ceramide. The results show that inhibition of nCDase activates GSK3β through dephosphorylation, and thus is required for the subsequent phosphorylation and degradation of β-catenin. Our findings show that inhibition of nCDase also inhibits the basal activation status of AKT, and we further establish that a constitutively active AKT (AKT T308D, S473D; AKTDD) reverses the effect of nCDase on β-catenin degradation. Functionally, the AKTDD mutant is able to overcome the growth suppressive effects of nCDase inhibition in CRC cells. Moreover, nCDase inhibition induces a growth delay of xenograft tumors from control cells, whereas xenograft tumors from constitutively active AKT cells become resistant to nCDase inhibition. Taken together, these results provide important mechanistic insight into how nCDase regulates cell proliferation. These findings demonstrate a heretofore unappreciated, but critical, role for nCDase in enabling/maintaining basal activation of AKT and also suggest that nCDase is a suitable novel target for colon cancer therapy.

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References

  1. Fakih MG. Metastatic colorectal cancer: current state and future directions. J Clin Oncol. 2015;33:1809–24.

    Article  CAS  Google Scholar 

  2. Stewart B, Wild C. World Cancer Report 2014. 1. Lyon: International Agency for Research on Cancer; 2014. p. 619.

    Google Scholar 

  3. Torre LA, Siegel RL, Ward EM, Jemal A. Global cancer incidence and mortality rates and trends—an update. Cancer Epidemiol Biomark. 2016;25:16–27.

    Article  Google Scholar 

  4. Yeang CH, McCormick F, Levine A. Combinatorial patterns of somatic gene mutations in cancer. FASEB J. 2008;22:2605–22.

    Article  CAS  Google Scholar 

  5. Hannun YA, Obeid LM. Principles of bioactive lipid signalling: lessons from sphingolipids. Nat Rev Mol Cell Biol. 2008;9:139–50.

    Article  CAS  Google Scholar 

  6. Hannun YA, Obeid LM. Many ceramides. J Biol Chem. 2011;286:27855–62.

    Article  CAS  Google Scholar 

  7. Nikolova-Karakashian MN, Rozenova KA. Ceramide in stress response. Adv Exp Med Biol. 2010;688:86–108.

    Article  CAS  Google Scholar 

  8. Kitatani K, Idkowiak-Baldys J, Hannun YA. The sphingolipid salvage pathway in ceramide metabolism and signaling. Cell Signal. 2008;20:1010–8.

    Article  CAS  Google Scholar 

  9. Futerman AH, Hannun YA. The complex life of simple sphingolipids. EMBO Rep. 2004;5:777–82.

    Article  CAS  Google Scholar 

  10. Mao C, Obeid LM. Ceramidases: regulators of cellular responses mediated by ceramide, sphingosine, and sphingosine-1-phosphate. Biochim Biophys Acta. 2008;1781:424–34.

    Article  CAS  Google Scholar 

  11. Kono M, Dreier JL, Ellis JM, Allende ML, Kalkofen DN, Sanders KM, et al. Neutral ceramidase encoded by the Asah2 gene is essential for the intestinal degradation of sphingolipids. J Biol Chem. 2006;281:7324–31.

    Article  CAS  Google Scholar 

  12. Garcia-Barros M, Coant N, Kawamori T, Wada M, Snider AJ, Truman JP, et al. Role of neutral ceramidase in colon cancer. FASEB J. 2016;12:4159–4171.

    Article  Google Scholar 

  13. Wu D, Pan W. GSK3: a multifaceted kinase in Wnt signaling. Trends Biochem Sci. 2010;35:161–8.

    Article  CAS  Google Scholar 

  14. MacDonald BT, Tamai K, He X. Wnt/beta-catenin signaling: components, mechanisms, and diseases. Dev Cell. 2009;17:9–26.

    Article  CAS  Google Scholar 

  15. Clevers H. Wnt/beta-catenin signaling in development and disease. Cell. 2006;127:469–80.

    Article  CAS  Google Scholar 

  16. Ilyas M, Tomlinson IP, Rowan A, Pignatelli M, Bodmer WF. Beta-catenin mutations in cell lines established from human colorectal cancers. Proc Natl Acad Sci USA. 1997;94:10330–4.

    Article  CAS  Google Scholar 

  17. Berg KCG, Eide PW, Eilertsen IA, Johannessen B, Bruun J, Danielsen SA, et al. Multiomics of 34 colorectal cancer cell lines—a resource for biomedical studies. Mol Cancer. 2017;16:116.

    Article  Google Scholar 

  18. Vivanco I, Sawyers CL. The phosphatidylinositol 3-kinase AKT pathway in human cancer. Nat Rev Cancer. 2002;2:489–501.

    Article  CAS  Google Scholar 

  19. Krycer JR, Sharpe LJ, Luu W, Brown AJ. The Akt-SREBP nexus: cell signaling meets lipid metabolism. Trends Endocrinol Metab. 2010;21:268–76.

    Article  CAS  Google Scholar 

  20. Muller EJ, Williamson L, Kolly C, Suter MM. Outside-in signaling through integrins and cadherins: a central mechanism to control epidermal growth and differentiation? J Invest Dermatol. 2008;128:501–16.

    Article  CAS  Google Scholar 

  21. Datta SR, Brunet A, Greenberg ME. Cellular survival: a play in three Akts. Genes Dev. 1999;13:2905–27.

    Article  CAS  Google Scholar 

  22. Sasaki AT, Firtel RA. Regulation of chemotaxis by the orchestrated activation of Ras, PI3K, and TOR. Eur J Cell Biol. 2006;85:873–95.

    Article  CAS  Google Scholar 

  23. Jiang BH, Liu LZ. AKT signaling in regulating angiogenesis. Curr Cancer Drug Targets. 2008;8:19–26.

    Article  CAS  Google Scholar 

  24. Yuan TL, Cantley LC. PI3K pathway alterations in cancer: variations on a theme. Oncogene. 2008;27:5497–510.

    Article  CAS  Google Scholar 

  25. Knowles MA, Platt FM, Ross RL, Hurst CD. Phosphatidylinositol 3-kinase (PI3K) pathway activation in bladder cancer. Cancer Metastas Rev. 2009;28:305–16.

    Article  CAS  Google Scholar 

  26. Hafsi S, Pezzino FM, Candido S, Ligresti G, Spandidos DA, Soua Z, et al. Gene alterations in the PI3K/PTEN/AKT pathway as a mechanism of drug-resistance (review). Int J Oncol. 2012;40:639–44.

    CAS  PubMed  Google Scholar 

  27. Wu BX, Zeidan YH, Hannun YA. Downregulation of neutral ceramidase by gemcitabine: implications for cell cycle regulation. Biochim Biophys Acta. 2009;1791:730–9.

    Article  CAS  Google Scholar 

  28. Ahmed D, Eide PW, Eilertsen IA, Danielsen SA, Eknaes M, Hektoen M, et al. Epigenetic and genetic features of 24 colon cancer cell lines. Oncogenesis. 2013;2:e71.

    Article  CAS  Google Scholar 

  29. Zhou H, Summers SA, Birnbaum MJ, Pittman RN. Inhibition of Akt kinase by cell-permeable ceramide and its implications for ceramide-induced apoptosis. J Biol Chem. 1998;273:16568–75.

    Article  CAS  Google Scholar 

  30. Rahman A, Thayyullathil F, Pallichankandy S, Galadari S. Hydrogen peroxide/ceramide/Akt signaling axis play a critical role in the antileukemic potential of sanguinarine. Free Radic Biol Med. 2016;96:273–89.

    Article  CAS  Google Scholar 

  31. Holland WL, Bikman BT, Wang LP, Yuguang G, Sargent KM, Bulchand S, et al. Lipid-induced insulin resistance mediated by the proinflammatory receptor TLR4 requires saturated fatty acid-induced ceramide biosynthesis in mice. J Clin Invest. 2011;121:1858–70.

    Article  CAS  Google Scholar 

  32. Stoica BA, Movsesyan VA, Lea PMt, Faden AI. Ceramide-induced neuronal apoptosis is associated with dephosphorylation of Akt, BAD, FKHR, GSK-3beta, and induction of the mitochondrial-dependent intrinsic caspase pathway. Mol Cell Neurosci. 2003;22:365–82.

    Article  CAS  Google Scholar 

  33. Dobrowsky RT, Kamibayashi C, Mumby MC, Hannun YA. Ceramide activates heterotrimeric protein phosphatase 2A. J Biol Chem. 1993;268:15523–30.

    CAS  PubMed  Google Scholar 

  34. Oaks J, Ogretmen B. Regulation of PP2A by sphingolipid metabolism and signaling. Front Oncol. 2014;4:388.

    PubMed  Google Scholar 

  35. Halder K, Banerjee S, Bose A, Majumder S, Majumdar S. Overexpressed PKCdelta downregulates the expression of PKCalpha in B16F10 melanoma: induction of apoptosis by PKCdelta via ceramide generation. PLoS One. 2014;9:e91656.

    Article  Google Scholar 

  36. Powell DJ, Hajduch E, Kular G, Hundal HS. Ceramide disables 3-phosphoinositide binding to the pleckstrin homology domain of protein kinase B (PKB)/Akt by a PKCzeta-dependent mechanism. Mol Cell Biol. 2003;23:7794–808.

    Article  CAS  Google Scholar 

  37. Symolon H, Bushnev A, Peng Q, Ramaraju H, Mays SG, Allegood JC, et al. Enigmol: a novel sphingolipid analogue with anticancer activity against cancer cell lines and in vivo models for intestinal and prostate cancer. Mol Cancer Ther. 2011;10:648–57.

    Article  CAS  Google Scholar 

  38. Kumar A, Pandurangan AK, Lu F, Fyrst H, Zhang M, Byun HS, et al. Chemopreventive sphingadienes downregulate Wnt signaling via a PP2A/Akt/GSK3beta pathway in colon cancer. Carcinogenesis. 2012;33:1726–35.

    Article  CAS  Google Scholar 

  39. Garcia-Barros M, Coant N, Truman JP, Snider AJ, Hannun YA. Sphingolipids in colon cancer. Biochim Biophys Acta. 2014;1841:773–82.

    Article  CAS  Google Scholar 

  40. Kawamori T, Kaneshiro T, Okumura M, Maalouf S, Uflacker A, Bielawski J, et al. Role for sphingosine kinase 1 in colon carcinogenesis. FASEB J. 2009;23:405–14.

    Article  CAS  Google Scholar 

  41. Snider AJ, Wu BX, Jenkins RW, Sticca JA, Kawamori T, Hannun YA, et al. Loss of neutral ceramidase increases inflammation in a mouse model of inflammatory bowel disease. Prostaglandins Other Lipid Mediat. 2012;99:124–30.

    Article  CAS  Google Scholar 

  42. Andre T, Boni C, Mounedji-Boudiaf L, Navarro M, Tabernero J, Hickish T, et al. Oxaliplatin, fluorouracil, and leucovorin as adjuvant treatment for colon cancer. N Engl J Med. 2004;350:2343–51.

    Article  CAS  Google Scholar 

  43. Andre T, Boni C, Navarro M, Tabernero J, Hickish T, Topham C, et al. Improved overall survival with oxaliplatin, fluorouracil, and leucovorin as adjuvant treatment in stage II or III colon cancer in the MOSAIC trial. J Clin Oncol. 2009;27:3109–16.

    Article  CAS  Google Scholar 

  44. Carrato A. Adjuvant treatment of colorectal cancer. Gastrointest Cancer Res. 2008;2:S42–46.

    PubMed  PubMed Central  Google Scholar 

  45. Verdaguer H, Tabernero J, Macarulla T. Ramucirumab in metastatic colorectal cancer: evidence to date and place in therapy. Ther Adv Med Oncol. 2016;8:230–42.

    Article  CAS  Google Scholar 

  46. Nelson VM, Benson AB 3rd. Status of targeted therapies in the adjuvant treatment of colon cancer. J Gastrointest Oncol. 2013;4:245–52.

    CAS  PubMed  PubMed Central  Google Scholar 

  47. Airola MV, Allen WJ, Pulkoski-Gross MJ, Obeid LM, Rizzo RC, Hannun YA. Structural basis for ceramide recognition and hydrolysis by human neutral ceramidase. Structure. 2015;23:1482–91.

    Article  CAS  Google Scholar 

  48. Clark J, Anderson KE, Juvin V, Smith TS, Karpe F, Wakelam MJ, et al. Quantification of PtdInsP3 molecular species in cells and tissues by mass spectrometry. Nat Methods. 2011;8:267–72.

    Article  CAS  Google Scholar 

  49. Anderson KE, Juvin V, Clark J, Stephens LR, Hawkins PT. Investigating the effect of arachidonate supplementation on the phosphoinositide content of MCF10a breast epithelial cells. Adv Biol Regul. 2016;62:18–24.

    Article  CAS  Google Scholar 

  50. Gandy KA, Canals D, Adada M, Wada M, Roddy P, Snider AJ, et al. Sphingosine 1-phosphate induces filopodia formation through S1PR2 activation of ERM proteins. Biochem J. 2013;449:661–72.

    Article  CAS  Google Scholar 

  51. Kim JH, Alfieri AA, Kim SH, Young CW. Potentiation of radiation effects on two murine tumors by lonidamine. Cancer Res. 1986;46:1120–3.

    CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by NCI grants CA172517 and CA97132. We acknowledge the support from the “Research Histology Core Laboratory, Department of Pathology, Stony Brook University”. We would like to thank Ayanna Lewis, MD for proof reading the manuscript.

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Authors and Affiliations

  1. Stony Brook Cancer Center, Stony Brook University, Stony Brook, NY, USA

    Nicolas Coant, Mónica García-Barros, Lina M. Obeid & Yusuf A. Hannun

  2. Signalling Programme, Babraham Institute, Babraham Research Campus, Cambridge, UK

    Qifeng Zhang

  3. Department of Medicine, Stony Brook University, Stony Brook, NY, USA

    Lina M. Obeid & Yusuf A. Hannun

  4. Northport VA Medical Center, Northport, NY, USA

    Lina M. Obeid

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Correspondence to Yusuf A. Hannun.

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All animal studies were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of Stony Brook University.

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Coant, N., García-Barros, M., Zhang, Q. et al. AKT as a key target for growth promoting functions of neutral ceramidase in colon cancer cells. Oncogene 37, 3852–3863 (2018). https://doi.org/10.1038/s41388-018-0236-x

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