Although numerous genetic mutations and amplifications have been identified in pancreatic cancer, much of the molecular pathogenesis of the disease remains undefined. While proteomic and transcriptomic analyses have been utilized to probe and characterize pancreatic tumors, lipidomic analyses have not been applied to identify perturbations in pancreatic cancer patient samples. Thus, we utilized a mass spectrometry-based lipidomic approach, focused towards the sphingolipid class of lipids, to quantify changes in human pancreatic cancer tumor and plasma specimens. Subgroup analysis revealed that patients with positive lymph node metastasis have a markedly higher level of ceramide species (C16:0 and C24:1) in their tumor specimens compared to pancreatic cancer patients without nodal disease or to patients with pancreatitis. Also of interest, ceramide metabolites, including phosphorylated (sphingosine- and sphinganine-1-phosphate) and glycosylated (cerebroside) species were elevated in the plasma, but not the pancreas, of pancreatic cancer patients with nodal disease. Analysis of plasma level of cytokine and growth factors revealed that IL-6, IL-8, CCL11 (eotaxin), EGF and IP10 (interferon inducible protein 10, CXCL10) were elevated in patients with positive lymph nodes metastasis, but that only IP10 and EGF directly correlated with several sphingolipid changes. Taken together, these data indicate that sphingolipid metabolism is altered in human pancreatic cancer and associated with advanced disease. Assessing plasma and/or tissue sphingolipids could potentially risk stratify patients in the clinical setting.
References
[1]
Tholey, R.; Sawicki, J.A.; Brody, J.R. Molecular-based and alternative therapies for pancreatic cancer: Looking “out of the box”. Cancer J. 2012, 18, 665–673, doi:10.1097/PPO.0b013e3182793ff6.
[2]
Pan, S.; Brentnall, T.A.; Kelly, K.; Chen, R. Tissue proteomics in pancreatic cancer study: Discovery, emerging technologies and challenges. Proteomics 2013, 13, 710–721, doi:10.1002/pmic.201200319.
Ogretmen, B.; Hannun, Y.A. Biologically active sphingolipids in cancer pathogenesis and treatment. Nat. Rev. Cancer 2004, 4, 604–616, doi:10.1038/nrc1411.
Ryland, L.K.; Fox, T.E.; Liu, X.; Loughran, T.P.; Kester, M. Dysregulation of sphingolipid metabolism in cancer. Cancer Biol. Ther. 2011, 11, 138–149, doi:10.4161/cbt.11.2.14624.
[7]
Barth, B.M.; Cabot, M.C.; Kester, M. Ceramide-based therapeutics for the treatment of cancer. Anticancer Agents Med. Chem. 2011, 11, 911–919, doi:10.2174/187152011797655177.
[8]
Furuya, H.; Shimizu, Y.; Kawamori, T. Sphingolipids in cancer. Cancer Metastasis Rev. 2011, 30, 567–576, doi:10.1007/s10555-011-9304-1.
[9]
Futerman, A.H.; Hannun, Y.A. The complex life of simple sphingolipids. EMBO Rep. 2004, 5, 777–782, doi:10.1038/sj.embor.7400208.
[10]
Bose, R.; Verheij, M.; Haimovitz-Friedman, A.; Scotto, K.; Fuks, Z.; Kolesnick, R. Ceramide synthase mediates daunorubicin-induced apoptosis: An alternative mechanism for generating death signals. Cell 1995, 82, 405–414, doi:10.1016/0092-8674(95)90429-8.
[11]
Perry, D.K.; Carton, J.; Shah, A.K.; Meredith, F.; Uhlinger, D.J.; Hannun, Y.A. Serine palmitoyltransferase regulates de novo ceramide generation during etoposide-induced apoptosis. J. Biol. Chem. 2000, 275, 9078–9084.
[12]
Garcia-Barros, M.; Paris, F.; Cordon-Cardo, C.; Lyden, D.; Rafii, S.; Haimovitz-Friedman, A.; Fuks, Z.; Kolesnick, R. Tumor response to radiotherapy regulated by endothelial cell apoptosis. Science 2003, 300, 1155–1159, doi:10.1126/science.1082504.
[13]
Fox, T.E.; Bewley, M.C.; Unrath, K.A.; Pedersen, M.M.; Anderson, R.E.; Jung, D.Y.; Jefferson, L.S.; Kim, J.K.; Bronson, S.K.; Flanagan, J.M.; et al. Circulating sphingolipid biomarkers in models of type 1 diabetes. J. Lipid Res. 2011, 52, 509–517, doi:10.1194/jlr.M010595.
[14]
Koybasi, S.; Senkal, C.E.; Sundararaj, K.; Spassieva, S.; Bielawski, J.; Osta, W.; Day, T.A.; Jiang, J.C.; Jazwinski, S.M.; Hannun, Y.A.; et al. Defects in cell growth regulation by C18:0-ceramide and longevity assurance gene 1 in human head and neck squamous cell carcinomas. J. Biol. Chem. 2004, 279, 44311–44319, doi:10.1074/jbc.M406920200.
[15]
Karahatay, S.; Thomas, K.; Koybasi, S.; Senkal, C.E.; Elojeimy, S.; Liu, X.; Bielawski, J.; Day, T.A.; Gillespie, M.B.; Sinha, D.; et al. Clinical relevance of ceramide metabolism in the pathogenesis of human head and neck squamous cell carcinoma (HNSCC): Attenuation of C(18)-ceramide in HNSCC tumors correlates with lymphovascular invasion and nodal metastasis. Cancer Lett. 2007, 256, 101–111, doi:10.1016/j.canlet.2007.06.003.
[16]
Schiffmann, S.; Sandner, J.; Birod, K.; Wobst, I.; Angioni, C.; Ruckhaberle, E.; Kaufmann, M.; Ackermann, H.; Lotsch, J.; Schmidt, H.; et al. Ceramide synthases and ceramide levels are increased in breast cancer tissue. Carcinogenesis 2009, 30, 745–752, doi:10.1093/carcin/bgp061.
[17]
Senkal, C.E.; Ponnusamy, S.; Bielawski, J.; Hannun, Y.A.; Ogretmen, B. Antiapoptotic roles of ceramide-synthase-6-generated C16-ceramide via selective regulation of the ATF6/CHOP arm of ER-stress-response pathways. FASEB J. 2010, 24, 296–308, doi:10.1096/fj.09-135087.
[18]
Hartmann, D.; Lucks, J.; Fuchs, S.; Schiffmann, S.; Schreiber, Y.; Ferreiros, N.; Merkens, J.; Marschalek, R.; Geisslinger, G.; Grosch, S. Long chain ceramides and very long chain ceramides have opposite effects on human breast and colon cancer cell growth. Int. J. Biochem. Cell. Biol. 2012, 44, 620–628, doi:10.1016/j.biocel.2011.12.019.
[19]
Gouaze-Andersson, V.; Cabot, M.C. Glycosphingolipids and drug resistance. Biochim. Biophys. Acta 2006, 1758, 2096–2103, doi:10.1016/j.bbamem.2006.08.012.
[20]
Gouaze-Andersson, V.; Yu, J.Y.; Kreitenberg, A.J.; Bielawska, A.; Giuliano, A.E.; Cabot, M.C. Ceramide and glucosylceramide upregulate expression of the multidrug resistance gene MDR1 in cancer cells. Biochim. Biophys. Acta 2007, 1771, 1407–1417, doi:10.1016/j.bbalip.2007.09.005.
[21]
Couvelard, A.; Hu, J.; Steers, G.; O’Toole, D.; Sauvanet, A.; Belghiti, J.; Bedossa, P.; Gatter, K.; Ruszniewski, P.; Pezzella, F. Identification of potential therapeutic targets by gene-expression profiling in pancreatic endocrine tumors. Gastroenterology 2006, 131, 1597–1610, doi:10.1053/j.gastro.2006.09.007.
[22]
Park, H.U.; Kim, J.W.; Kim, G.E.; Bae, H.I.; Crawley, S.C.; Yang, S.C.; Gum, J.R., Jr.; Batra, S.K.; Rousseau, K.; Swallow, D.M.; et al. Aberrant expression of MUC3 and MUC4 membrane-associated mucins and sialyl Le(x) antigen in pancreatic intraepithelial neoplasia. Pancreas 2003, 26, e48–e54, doi:10.1097/00006676-200304000-00022.
[23]
Pfeffer, F.; Koczan, D.; Adam, U.; Benz, S.; von Dobschuetz, E.; Prall, F.; Nizze, H.; Thiesen, H.J.; Hopt, U.T.; Lobler, M. Expression of connexin26 in islets of Langerhans is associated with impaired glucose tolerance in patients with pancreatic adenocarcinoma. Pancreas 2004, 29, 284–290, doi:10.1097/00006676-200411000-00007.
[24]
Sato, N.; Fukushima, N.; Maitra, A.; Iacobuzio-Donahue, C.A.; van Heek, N.T.; Cameron, J.L.; Yeo, C.J.; Hruban, R.H.; Goggins, M. Gene expression profiling identifies genes associated with invasive intraductal papillary mucinous neoplasms of the pancreas. Am. J. Pathol. 2004, 164, 903–914, doi:10.1016/S0002-9440(10)63178-1.
[25]
Hamanaka, Y.; Hamanaka, S.; Suzuki, M. Sialyl Lewis(a) ganglioside in pancreatic cancer tissue correlates with the serum CA 19-9 level. Pancreas 1996, 13, 160–165, doi:10.1097/00006676-199608000-00007.
[26]
Saddoughi, S.A.; Song, P.; Ogretmen, B. Roles of bioactive sphingolipids in cancer biology and therapeutics. Subcell Biochem. 2008, 49, 413–440, doi:10.1007/978-1-4020-8831-5_16.
[27]
Nava, V.E.; Hobson, J.P.; Murthy, S.; Milstien, S.; Spiegel, S. Sphingosine kinase type 1 promotes estrogen-dependent tumorigenesis of breast cancer MCF-7 cells. Exp. Cell. Res. 2002, 281, 115–127, doi:10.1006/excr.2002.5658.
[28]
Van Brocklyn, J.; Letterle, C.; Snyder, P.; Prior, T. Sphingosine-1-phosphate stimulates human glioma cell proliferation through Gi-coupled receptors: Role of ERK MAP kinase and phosphatidylinositol 3-kinase beta. Cancer Lett. 2002, 181, 195–204, doi:10.1016/S0304-3835(02)00050-2.
[29]
Sarkar, S.; Maceyka, M.; Hait, N.C.; Paugh, S.W.; Sankala, H.; Milstien, S.; Spiegel, S. Sphingosine kinase 1 is required for migration, proliferation and survival of MCF-7 human breast cancer cells. FEBS Lett. 2005, 579, 5313–5317, doi:10.1016/j.febslet.2005.08.055.
[30]
Ponnusamy, S.; Selvam, S.P.; Mehrotra, S.; Kawamori, T.; Snider, A.J.; Obeid, L.M.; Shao, Y.; Sabbadini, R.; Ogretmen, B. Communication between host organism and cancer cells is transduced by systemic sphingosine kinase 1/sphingosine 1-phosphate signalling to regulate tumour metastasis. EMBO Mol. Med. 2012, 4, 761–775, doi:10.1002/emmm.201200244.
[31]
Fox, T.E.; Bewley, M.C.; Unrath, K.A.; Pedersen, M.M.; Anderson, R.E.; Jung, D.Y.; Jefferson, L.S.; Kim, J.K.; Bronson, S.K.; Flanagan, J.M.; et al. Circulating sphingolipid biomarkers in models of type 1 diabetes. J. Lipid Res. 2011, 52, 509–517, doi:10.1194/jlr.M010595.
Long, J.S.; Fujiwara, Y.; Edwards, J.; Tannahill, C.L.; Tigyi, G.; Pyne, S.; Pyne, N.J. Sphingosine 1-phosphate receptor 4 uses HER2 (ERBB2) to regulate extracellular signal regulated kinase-1/2 in MDA-MB-453 breast cancer cells. J. Biol. Chem. 2010, 285, 35957–35966.
[34]
Shida, D.; Fang, X.; Kordula, T.; Takabe, K.; Lepine, S.; Alvarez, S.E.; Milstien, S.; Spiegel, S. Cross-talk between LPA1 and epidermal growth factor receptors mediates up-regulation of sphingosine kinase 1 to promote gastric cancer cell motility and invasion. Cancer Res. 2008, 68, 6569–6577, doi:10.1158/0008-5472.CAN-08-0411.
[35]
Doll, F.; Pfeilschifter, J.; Huwiler, A. The epidermal growth factor stimulates sphingosine kinase-1 expression and activity in the human mammary carcinoma cell line MCF7. Biochim. Biophys. Acta 2005, 1738, 72–81, doi:10.1016/j.bbalip.2005.12.001.
[36]
Evans, A.; Costello, E. The role of inflammatory cells in fostering pancreatic cancer cell growth and invasion. Front. Physiol. 2012, 3, 270.
