Sphingosine kinase 2 deficiency increases proliferation and migration of renal mouse mesangial cells and fibroblasts
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Stephanie Schwalm
, Tankica Maneva Timcheva
Abstract
Both of the sphingosine kinase (SK) subtypes SK-1 and SK-2 catalyze the production of the bioactive lipid molecule sphingosine 1-phosphate (S1P). However, the subtype-specific cellular functions are largely unknown. In this study, we investigated the cellular function of SK-2 in primary mouse renal mesangial cells (mMC) and embryonic fibroblasts (MEF) from wild-type C57BL/6 or SK-2 knockout (SK2ko) mice. We found that SK2ko cells displayed a significantly higher proliferative and migratory activity when compared to wild-type cells, with concomitant increased cellular activities of the classical extracellular signal regulated kinase (ERK) and PI3K/Akt cascades, and of the small G protein RhoA. Furthermore, we detected an upregulation of SK-1 protein and S1P3 receptor mRNA expression in SK-2ko cells. The MEK inhibitor U0126 and the S1P1/3 receptor antagonist VPC23019 blocked the increased migration of SK-2ko cells. Additionally, S1P3ko mesangial cells showed a reduced proliferative behavior and reduced migration rate upon S1P stimulation, suggesting a crucial involvement of the S1P3 receptor. In summary, our data demonstrate that SK-2 exerts suppressive effects on cell growth and migration in renal mesangial cells and fibroblasts, and that therapeutic targeting of SKs for treating proliferative diseases requires subtype-selective inhibitors.
Acknowledgments
This work was supported by the Swiss National Science Foundation (3100A0–111806), and the German Research Foundation (SFB1039/TP02). We thank Isolde Römer and Simone Albert for technical assistance.
References
Alemany, R., van Koppen, C.J., Danneberg, K., Ter Braak, M., and Meyer Zu Heringdorf, D. (2007). Regulation and functional roles of sphingosine kinases. Naunyn-Schmiedeberg’s Arch. Pharmacol. 374, 413–428.Suche in Google Scholar
An, S., Zheng, Y., and Bleu, T. (2000). Sphingosine 1-phosphate-induced cell proliferation, survival, and related signaling events mediated by G protein-coupled receptors Edg3 and Edg5. J. Biol. Chem. 275, 288–296.10.1074/jbc.275.1.288Suche in Google Scholar PubMed
Antoon, J.W., White, M.D., Meacham, W.D., Slaughter, E.M., Muir, S.E., Elliott, S., Rhodes, L.V., Ashe, H.B., Wiese, T.E., Smith, C.D., et al. (2010). Antiestrogenic effects of the novel sphingosine kinase-2 inhibitor ABC294640. Endocrinology 151, 5124–5135.10.1210/en.2010-0420Suche in Google Scholar PubMed PubMed Central
Antoon, J.W., White, M.D., Slaughter, E.M., Driver, J.L., Khalili, H.S., Elliott, S., Smith, C.D., Burow, M.E., and Beckman, B.S. (2011). Targeting NF-kB mediated breast cancer chemoresistance through selective inhibition of sphingosine kinase-2. Cancer Biol. Ther. 11, 678–689.10.4161/cbt.11.7.14903Suche in Google Scholar PubMed PubMed Central
Arikawa, K., Takuwa, N., Yamaguchi, H., Sugimoto, N., Kitayama, J., Nagawa, H., Takehara, K., and Takuwa, Y. (2003). Ligand-dependent inhibition of B16 melanoma cell migration and invasion via endogenous S1P2 G protein-coupled receptor. Requirement of inhibition of cellular RAC activity. J. Biol. Chem. 278, 32841–32851.10.1074/jbc.M305024200Suche in Google Scholar PubMed
Baker, D.A., Eudaly, J., Smith, C.D., Obeid, L.M., and Gilkeson, G.S. (2013). Impact of sphingosine kinase 2 deficiency on the development of TNF-a-induced inflammatory arthritis. Rheumatol. Int. 33, 2677–2681.10.1007/s00296-012-2493-2Suche in Google Scholar PubMed PubMed Central
Bessa, C., Pereira, C., Leao, M., Maciel, C., Gomes, S., Goncalves, J., Corte-Real, M., Costa, V., and Saraiva, L. (2013). Using yeast to uncover the regulation of protein kinase Cdelta by ceramide. FEMS Yeast Res. 13, 700–705.10.1111/1567-1364.12069Suche in Google Scholar PubMed
Calise, S., Blescia, S., Cencetti, F., Bernacchioni, C., Donati, C., and Bruni, P. (2012). Sphingosine 1-phosphate stimulates proliferation and migration of satellite cells: role of S1P receptors. Biochim. Biophys. Acta. 1823, 439–450.10.1016/j.bbamcr.2011.11.016Suche in Google Scholar PubMed
Conner, D.A. (2001). Mouse embryo fibroblast (MEF) feeder cell preparation. Curr. Protoc. Mol. Biol. Chapter 23, Unit 23 22.10.1002/0471142727.mb2302s51Suche in Google Scholar PubMed
Döll, F., Pfeilschifter, J., and Huwiler, A. (2005). The epidermal growth factor stimulates sphingosine kinase-1 expression and activity in the human mammary carcinoma cell line MCF7. Biochim. Biophys. Acta. 1738, 72–81.10.1016/j.bbalip.2005.12.001Suche in Google Scholar PubMed
Döll, F., Pfeilschifter, J., and Huwiler, A. (2007). Prolactin upregulates sphingosine kinase-1 expression and activity in the human breast cancer cell line MCF7 and triggers enhanced proliferation and migration. Endocr. Relat. Cancer 14, 325–335.10.1677/ERC-06-0050Suche in Google Scholar PubMed
Favata, M.F., Horiuchi, K.Y., Manos, E.J., Daulerio, A.J., Stradley, D.A., Feeser, W.S., Van Dyk, D.E., Pitts, W.J., Earl, R.A., Hobbs, F., et al. (1998). Identification of a novel inhibitor of mitogen-activated protein kinase kinase. J. Biol. Chem. 273, 18623–18632.10.1074/jbc.273.29.18623Suche in Google Scholar PubMed
French, K.J., Schrecengost, R.S., Lee, B.D., Zhuang, Y., Smith, S.N., Eberly, J.L., Yun, J.K., and Smith, C.D. (2003). Discovery and evaluation of inhibitors of human sphingosine kinase. Cancer Res. 63, 5962–5969.Suche in Google Scholar
French, K.J., Upson, J.J., Keller, S.N., Zhuang, Y., Yun, J.K., and Smith, C.D. (2006). Antitumor activity of sphingosine kinase inhibitors. J. Pharmacol. Exp. Ther. 318, 596–603.10.1124/jpet.106.101345Suche in Google Scholar PubMed
French, K.J., Zhuang, Y., Maines, L.W., Gao, P., Wang, W., Beljanski, V., Upson, J.J., Green, C.L., Keller, S.N., and Smith, C.D. (2010). Pharmacology and antitumor activity of ABC294640, a selective inhibitor of sphingosine kinase-2. J. Pharmacol. Exp. Ther. 333, 129–139.10.1124/jpet.109.163444Suche in Google Scholar PubMed PubMed Central
Goparaju, S.K., Jolly, P.S., Watterson, K.R., Bektas, M., Alvarez, S., Sarkar, S., Mel, L., Ishii, I., Chun, J., Milstien, S., et al. (2005). The S1P2 receptor negatively regulates platelet-derived growth factor-induced motility and proliferation. Mol. Cell. Biol. 25, 4237–4249.10.1128/MCB.25.10.4237-4249.2005Suche in Google Scholar PubMed PubMed Central
Hait, N.C., Bellamy, A., Milstien, S., Kordula, T., and Spiegel, S. (2007). Sphingosine kinase type 2 activation by ERK-mediated phosphorylation. J. Biol. Chem. 282, 12058–12065.10.1074/jbc.M609559200Suche in Google Scholar PubMed
Hall, A. (1990). The cellular functions of small GTP-binding proteins. Science 249, 635–640.10.1126/science.2116664Suche in Google Scholar PubMed
Hofmann, L.P., Ren, S., Schwalm, S., Pfeilschifter, J., and Huwiler, A. (2008). Sphingosine kinase 1 and 2 regulate the capacity of mesangial cells to resist apoptotic stimuli in an opposing manner. Biol. Chem. 389, 1399–1407.10.1515/BC.2008.160Suche in Google Scholar PubMed
Huwiler, A., Brunner, J., Hummel, R., Vervoordeldonk, M., Stabel, S., van den Bosch, H., and Pfeilschifter, J. (1996). Ceramide-binding and activation defines protein kinase c-Raf as a ceramide-activated protein kinase. Proc. Natl. Acad. Sci. USA 93, 6959–6963.10.1073/pnas.93.14.6959Suche in Google Scholar PubMed PubMed Central
Huwiler, A., Doll, F., Ren, S., Klawitter, S., Greening, A., Romer, I., Bubnova, S., Reinsberg, L., and Pfeilschifter, J. (2006). Histamine increases sphingosine kinase-1 expression and activity in the human arterial endothelial cell line EA.hy 926 by a PKC-alpha-dependent mechanism. Biochim. Biophys. Acta. 1761, 367–376.10.1016/j.bbalip.2006.02.007Suche in Google Scholar PubMed
Huwiler, A., Fabbro, D., and Pfeilschifter, J. (1998). Selective ceramide binding to protein kinase C-a and -d isoenzymes in renal mesangial cells. Biochemistry 37, 14556–14562.10.1021/bi981401iSuche in Google Scholar PubMed
Huwiler, A., Kolter, T., Pfeilschifter, J., and Sandhoff, K. (2000). Physiology and pathophysiology of sphingolipid metabolism and signaling. Biochim. Biophys. Acta. 1485, 63–99.10.1016/S1388-1981(00)00042-1Suche in Google Scholar
Huwiler, A. and Pfeilschifter, J. (2006). Altering the sphingosine-1-phosphate/ceramide balance: a promising approach for tumor therapy. Curr. Pharm. Des. 12, 4625–4635.10.2174/138161206779010422Suche in Google Scholar
Igarashi, N., Okada, T., Hayashi, S., Fujita, T., Jahangeer, S., and Nakamura, S. (2003). Sphingosine kinase 2 is a nuclear protein and inhibits DNA synthesis. J. Biol. Chem. 278, 46832–46839.10.1074/jbc.M306577200Suche in Google Scholar
Ishii, I., Friedman, B., Ye, X., Kawamura, S., McGiffert, C., Contos, J.J., Kingsbury, M.A., Zhang, G., Brown, J.H., and Chun, J. (2001). Selective loss of sphingosine 1-phosphate signaling with no obvious phenotypic abnormality in mice lacking its G protein-coupled receptor, LP(B3)/EDG-3. J. Biol. Chem. 276, 33697–33704.10.1074/jbc.M104441200Suche in Google Scholar
Jo, S.K., Bajwa, A., Ye, H., Vergis, A.L., Awad, A.S., Kharel, Y., Lynch, K.R., and Okusa, M.D. (2009). Divergent roles of sphingosine kinases in kidney ischemia-reperfusion injury. Kidney Int. 75, 167–175.10.1038/ki.2008.400Suche in Google Scholar
Katsuma, S., Hada, Y., Ueda, T., Shiojima, S., Hirasawa, A., Tanoue, A., Takagaki, K., Ohgi, T., Yano, J., and Tsujimoto, G. (2002). Signalling mechanisms in sphingosine 1-phosphate-promoted mesangial cell proliferation. Genes Cells 7, 1217–1230.10.1046/j.1365-2443.2002.00594.xSuche in Google Scholar
Keely, P.J., Conklin, M.W., Gehler, S., Ponik, S.M., and Provenzano, P.P. (2007). Investigating integrin regulation and signaling events in three-dimensional systems. Methods Enzymol. 426, 27–45.10.1016/S0076-6879(07)26002-1Suche in Google Scholar
Klawitter, S., Hofmann, L.P., Pfeilschifter, J., and Huwiler, A. (2007). Extracellular nucleotides induce migration of renal mesangial cells by upregulating sphingosine kinase-1 expression and activity. Br. J. Pharmacol. 150, 271–280.10.1038/sj.bjp.0706983Suche in Google Scholar PubMed PubMed Central
Kunkel, G.T., Maceyka, M., Milstien, S., and Spiegel, S. (2013). Targeting the sphingosine-1-phosphate axis in cancer, inflammation and beyond. Nat. Rev. Drug Discov. 12, 688–702.10.1038/nrd4099Suche in Google Scholar PubMed PubMed Central
Liang, J., Nagahashi, M., Kim, E.Y., Harikumar, K.B., Yamada, A., Huang, W.C., Hait, N.C., Allegood, J.C., Price, M.M., Avni, D., et al. (2013). Sphingosine-1-phosphate links persistent STAT3 activation, chronic intestinal inflammation, and development of colitis-associated cancer. Cancer Cell 23, 107–120.10.1016/j.ccr.2012.11.013Suche in Google Scholar PubMed PubMed Central
Liu, H., Sugiura, M., Nava, V.E., Edsall, L.C., Kono, K., Poulton, S., Milstien, S., Kohama, T., and Spiegel, S. (2000a). Molecular cloning and functional characterization of a novel mammalian sphingosine kinase type 2 isoform. J. Biol. Chem. 275, 19513–19520.10.1074/jbc.M002759200Suche in Google Scholar
Liu, Y., Wada, R., Yamashita, T., Mi, Y., Deng, C.X., Hobson, J.P., Rosenfeldt, H.M., Nava, V.E., Chae, S.S., Lee, M.J., et al. (2000b). Edg-1, the G protein-coupled receptor for sphingosine-1-phosphate, is essential for vascular maturation. J. Clin. Invest. 106, 951–961.10.1172/JCI10905Suche in Google Scholar
Liu, H., Toman, R.E., Goparaju, S.K., Maceyka, M., Nava, V.E., Sankala, H., Payne, S.G., Bektas, M., Ishii, I., Chun, J., et al. (2003). Sphingosine kinase type 2 is a putative BH3-only protein that induces apoptosis. J. Biol. Chem. 278, 40330–40336.10.1074/jbc.M304455200Suche in Google Scholar
Muller, G., Ayoub, M., Storz, P., Rennecke, J., Fabbro, D., and Pfizenmaier, K. (1995). PKC zeta is a molecular switch in signal transduction of TNF-a, bifunctionally regulated by ceramide and arachidonic acid. EMBO J. 14, 1961–1969.10.1002/j.1460-2075.1995.tb07188.xSuche in Google Scholar
O’Connor, K. and Chen, M. (2013). Dynamic functions of RhoA in tumor cell migration and invasion. Small GTPases 4, 141–147.10.4161/sgtp.25131Suche in Google Scholar
Okada, T., Ding, G., Sonoda, H., Kajimoto, T., Haga, Y., Khosrowbeygi, A., Gao, S., Miwa, N., Jahangeer, S., and Nakamura, S. (2005). Involvement of N-terminal-extended form of sphingosine kinase 2 in serum-dependent regulation of cell proliferation and apoptosis. J. Biol. Chem. 280, 36318–36325.10.1074/jbc.M504507200Suche in Google Scholar
Okamoto, H., Takuwa, N., Yokomizo, T., Sugimoto, N., Sakurada, S., Shigematsu, H., and Takuwa, Y. (2000). Inhibitory regulation of Rac activation, membrane ruffling, and cell migration by the G protein-coupled sphingosine-1-phosphate receptor EDG5 but not EDG1 or EDG3. Mol. Cell. Biol. 20, 9247–9261.10.1128/MCB.20.24.9247-9261.2000Suche in Google Scholar
Osada, M., Yatomi, Y., Ohmori, T., Ikeda, H., and Ozaki, Y. (2002). Enhancement of sphingosine 1-phosphate-induced migration of vascular endothelial cells and smooth muscle cells by an EDG-5 antagonist. Biochem. Biophys. Res. Commun. 299, 483–487.10.1016/S0006-291X(02)02671-2Suche in Google Scholar
Paik, J.H., Chae, S., Lee, M.J., Thangada, S., and Hla, T. (2001). Sphingosine 1-phosphate-induced endothelial cell migration requires the expression of EDG-1 and EDG-3 receptors and Rho-dependent activation of avb3- and b1-containing integrins. J. Biol. Chem. 276, 11830–11837.10.1074/jbc.M009422200Suche in Google Scholar PubMed
Pertz, O., Hodgson, L., Klemke, R.L., and Hahn, K.M. (2006). Spatiotemporal dynamics of RhoA activity in migrating cells. Nature 440, 1069–1072.10.1038/nature04665Suche in Google Scholar PubMed
Pyne, N.J. and Pyne, S. (2010). Sphingosine 1-phosphate and cancer. Nat. Rev. Cancer 10, 489–503.10.1038/nrc2875Suche in Google Scholar
Rölz, W., Xin, C., Ren, S., Pfeilschifter, J., and Huwiler, A. (2002). Interleukin-1 inhibits angiotensin II-stimulated protein kinase B pathway in renal mesangial cells via the inducible nitric oxide synthase. Eur. J. Pharmacol. 442, 195–203.10.1016/S0014-2999(02)01545-5Suche in Google Scholar
Ruckhäberle, E., Rody, A., Engels, K., Gaetje, R., von Minckwitz, G., Schiffmann, S., Grösch, S., Geisslinger, G., Holtrich, U., Karn, T., et al. (2008). Microarray analysis of altered sphingolipid metabolism reveals prognostic significance of sphingosine kinase 1 in breast cancer. Breast Cancer Res. Treat. 112, 41–52.10.1007/s10549-007-9836-9Suche in Google Scholar PubMed
Shida, D., Takabe, K., Kapitonov, D., Milstien, S., and Spiegel, S. (2008). Targeting SphK1 as a new strategy against cancer. Curr. Drug Targets 9, 662–673.10.2174/138945008785132402Suche in Google Scholar PubMed PubMed Central
Vivanco, I. and Sawyers, C.L. (2002). The phosphatidylinositol 3-Kinase AKT pathway in human cancer. Nat. Rev. Cancer 2, 489–501.10.1038/nrc839Suche in Google Scholar PubMed
vom Dorp, F., Sanders, H., Boergermann, C., Lummen, G., Rubben, H., Jakobs, K.H., and Schmidt, M. (2011). Inhibition of Rho-kinase abrogates migration of human transitional cell carcinoma cells: results of an in vitro study. Urol. Int. 86, 220–227.10.1159/000321271Suche in Google Scholar PubMed
Wymann, M.P., Bulgarelli-Leva, G., Zvelebil, M.J., Pirola, L., Vanhaesebroeck, B., Waterfield, M.D., and Panayotou, G. (1996). Wortmannin inactivates phosphoinositide 3-kinase by covalent modification of Lys-802, a residue involved in the phosphate transfer reaction. Mol. Cell Biol. 16, 1722–1733.10.1128/MCB.16.4.1722Suche in Google Scholar PubMed PubMed Central
Zhang, W., Zhao, J., Lee, J.F., Gartung, A., Jawadi, H., Lambiv, W.L., Honn, K.V., and Lee, M.J. (2013). ETS-1-mediated transcriptional up-regulation of CD44 is required for sphingosine-1-phosphate receptor subtype 3-stimulated chemotaxis. J. Biol. Chem. 288, 32126–32137.10.1074/jbc.M113.495218Suche in Google Scholar PubMed PubMed Central
©2015 by De Gruyter
Artikel in diesem Heft
- Frontmatter
- Guest Editorial
- Highlight: Molecular Medicine of Sphingolipids
- HIGHLIGHT: MOLECULAR MEDICINE OF SPHINGOLIPIDS
- The role of serum amyloid A and sphingosine-1-phosphate on high-density lipoprotein functionality
- Sphingolipids in viral infection
- Tackling the biophysical properties of sphingolipids to decipher their biological roles
- Ceramide and sphingosine in pulmonary infections
- Molecular mechanisms of erythrocyte aging
- Sphingolipids in liver injury, repair and regeneration
- Ultrasound-stimulated microbubble enhancement of radiation response
- Innate immune responses in the brain of sphingolipid lysosomal storage diseases
- Novel mechanisms of action of classical chemotherapeutic agents on sphingolipid pathways
- The role of sphingolipids in endothelial barrier function
- The effect of altered sphingolipid acyl chain length on various disease models
- Secretory sphingomyelinase in health and disease
- Preclinical development of a C6-ceramide NanoLiposome, a novel sphingolipid therapeutic
- Sphingomyelin breakdown in T cells: role in activation, effector functions and immunoregulation
- The molecular medicine of acid ceramidase
- Caenorhabditis elegans as a model to study sphingolipid signaling
- S1PR4 is required for plasmacytoid dendritic cell differentiation
- Antinociceptive effects of FTY720 during trauma-induced neuropathic pain are mediated by spinal S1P receptors
- Subcellular distribution of FTY720 and FTY720-phosphate in immune cells – another aspect of Fingolimod action relevant for therapeutic application
- Downregulation of sphingosine 1-phosphate (S1P) receptor 1 by dexamethasone inhibits S1P-induced mesangial cell migration
- Sphingosine kinase 2 deficiency increases proliferation and migration of renal mouse mesangial cells and fibroblasts
- Obituary
- The life and work of Dr. Robert Bittman (1942–2014)
Artikel in diesem Heft
- Frontmatter
- Guest Editorial
- Highlight: Molecular Medicine of Sphingolipids
- HIGHLIGHT: MOLECULAR MEDICINE OF SPHINGOLIPIDS
- The role of serum amyloid A and sphingosine-1-phosphate on high-density lipoprotein functionality
- Sphingolipids in viral infection
- Tackling the biophysical properties of sphingolipids to decipher their biological roles
- Ceramide and sphingosine in pulmonary infections
- Molecular mechanisms of erythrocyte aging
- Sphingolipids in liver injury, repair and regeneration
- Ultrasound-stimulated microbubble enhancement of radiation response
- Innate immune responses in the brain of sphingolipid lysosomal storage diseases
- Novel mechanisms of action of classical chemotherapeutic agents on sphingolipid pathways
- The role of sphingolipids in endothelial barrier function
- The effect of altered sphingolipid acyl chain length on various disease models
- Secretory sphingomyelinase in health and disease
- Preclinical development of a C6-ceramide NanoLiposome, a novel sphingolipid therapeutic
- Sphingomyelin breakdown in T cells: role in activation, effector functions and immunoregulation
- The molecular medicine of acid ceramidase
- Caenorhabditis elegans as a model to study sphingolipid signaling
- S1PR4 is required for plasmacytoid dendritic cell differentiation
- Antinociceptive effects of FTY720 during trauma-induced neuropathic pain are mediated by spinal S1P receptors
- Subcellular distribution of FTY720 and FTY720-phosphate in immune cells – another aspect of Fingolimod action relevant for therapeutic application
- Downregulation of sphingosine 1-phosphate (S1P) receptor 1 by dexamethasone inhibits S1P-induced mesangial cell migration
- Sphingosine kinase 2 deficiency increases proliferation and migration of renal mouse mesangial cells and fibroblasts
- Obituary
- The life and work of Dr. Robert Bittman (1942–2014)