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Role of Rho kinase in sphingosine 1-phosphate-mediated endothelial and smooth muscle cell migration and differentiation

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The role of sphingosine 1-phosphate (S1P)-induced Rho kinase (ROCK) activation in the angiogenic responses of pulmonary artery-derived endothelial cells (PAEC) and smooth muscle cells (PASMC) was examined. S1P, a biologically active phospholipid that regulates angiogenesis, promoted PAEC chemotaxis and capillary morphogenesis; furthermore, this activity was unaltered by pretreatment with the pharmacological inhibitor of ROCK, H1152. In contrast, S1P (500 nM) significantly inhibited spontaneous PASMC chemotaxis and differentiation; however, this inhibition was eradicated upon H1152 pretreatment. Similarly, PASMCs transfected with ROCK II siRNA diminished S1P-induced inhibition of the development of multi-cellular structures. Analysis by RT-PCR identified the presence of S1P1 and S1P3 receptors on both PAECs and PASMCs, while S1P2 receptor expression was confined to only PASMCs. Consistent with this observation, the S1P1 and S1P3 receptor antagonist, VPC23019, virtually abolished the S1P-initiated PAEC differentiation but did not impede the S1P-induced inhibition of PASMC differentiation. However, the S1P2 receptor antagonist, JTE013, had no effect on S1P-mediated differentiation of PAECs but abolished the S1P-induced inhibition of PASMC function. Co-cultured endothelial and smooth muscle cells differentiated into “neovascular-like” networks, which were significantly inhibited by S1P. The inhibition of co-culture differentiation in both PAECs and PASMCs was negated by H1152 pretreatment. However, when smooth muscle cells were added to S1P-initiated endothelial cell networks, additional S1P treatment did not inhibit the cellular networks generated by these cells. In conclusion, S1P-induced PAEC angiogenic responses are regulated by S1P1 and/or S1P3 receptors independent of Rho kinase activation, whereas S1P2 receptor-mediated curtailment of PASMC function by S1P.

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References

  1. Folkman J, Shing Y (1992) Angiogenesis. J Biol Chem 267:10931–10934

    CAS  PubMed  Google Scholar 

  2. Harvey K, Siddiqui RA, Sliva D et al (2002) Serum factors involved in human microvascular endothelial cell morphogenesis. J Lab Clin Med 140:188–198

    Article  CAS  PubMed  Google Scholar 

  3. Liu ZJ, Snyder R, Soma A et al (2003) VEGF-A and alphaVbeta3 integrin synergistically rescue angiogenesis via N-Ras and PI3-K signaling in human microvascular endothelial cells. FASEB J 17:1931–1933

    CAS  PubMed  Google Scholar 

  4. Van Belle E, Witzenbichler B, Chen D et al (1998) Potentiated angiogenic effect of scatter factor/hepatocyte growth factor via induction of vascular endothelial growth factor: the case for paracrine amplification of angiogenesis. Circulation 97:381–390

    PubMed  Google Scholar 

  5. Sengupta S, Gherardi E, Sellers LA et al (2003) Hepatocyte growth factor/scatter factor can induce angiogenesis independently of vascular endothelial growth factor. Arterioscler Thromb Vasc Biol 23:69–75

    Article  CAS  PubMed  Google Scholar 

  6. Zlot C, Ingle G, Hongo J et al (2003) Stanniocalcin 1 is an autocrine modulator of endothelial angiogenic responses to hepatocyte growth factor. J Biol Chem 278:47654–47659

    Article  CAS  PubMed  Google Scholar 

  7. Liu F, Verin AD, Wang P et al (2001) Differential regulation of sphingosine-1-phosphate- and VEGF-induced endothelial cell chemotaxis. Involvement of G(ialpha2)-linked Rho kinase activity. Am J Respir Cell Mol Biol 24:711–719

    CAS  PubMed  Google Scholar 

  8. Lee MJ, Thangada S, Claffey KP et al (1999) Vascular endothelial cell adherens junction assembly and morphogenesis induced by sphingosine-1-phosphate. Cell 99:301–312

    Article  CAS  PubMed  Google Scholar 

  9. Boguslawski G, Grogg JR, Welch Z et al (2002) Migration of vascular smooth muscle cells induced by sphingosine 1-phosphate and related lipids: potential role in the angiogenic response. Exp Cell Res 274:264–274

    Article  CAS  PubMed  Google Scholar 

  10. Wang F, Van Brocklyn JR, Hobson JP et al (1999) Sphingosine 1-phosphate stimulates cell migration through a G(i)-coupled cell surface receptor. Potential involvement in angiogenesis. J Biol Chem 274:35343–35350

    Article  CAS  PubMed  Google Scholar 

  11. Pyne S, Pyne NJ (2000) Sphingosine 1-phosphate signalling in mammalian cells. Biochem J 349:385–402

    Article  CAS  PubMed  Google Scholar 

  12. Pyne S, Pyne N (2000) Sphingosine 1-phosphate signalling via the endothelial differentiation gene family of G-protein-coupled receptors. Pharmacol Ther 88:115–131

    Article  CAS  PubMed  Google Scholar 

  13. Kluk MJ, Hla T (2002) Signaling of sphingosine-1-phosphate via the S1P/EDG-family of G-protein-coupled receptors. Biochim Biophys Acta 1582:72–80

    CAS  PubMed  Google Scholar 

  14. English D, Welch Z, Kovala AT et al (2000) Sphingosine 1-phosphate released from platelets during clotting accounts for the potent endothelial cell chemotactic activity of blood serum and provides a novel link between hemostasis and angiogenesis. FASEB J 14:2255–2265

    Article  CAS  PubMed  Google Scholar 

  15. Tolle M, Levkau B, Kleuser B et al (2007) Sphingosine-1-phosphate and FTY720 as anti-atherosclerotic lipid compounds. Eur J Clin Invest 37:171–179

    Article  CAS  PubMed  Google Scholar 

  16. Yatomi Y (2006) Sphingosine 1-phosphate in vascular biology: possible therapeutic strategies to control vascular diseases. Curr Pharm Des 12:575–587

    Article  CAS  PubMed  Google Scholar 

  17. Nofer J-R, van der Giet M, Tolle M et al (2004) HDL induces NO-dependent vasorelaxation via the lysophospholipid receptor S1P3 [see comment]. J Clin Invest 113:569–581

    CAS  PubMed  Google Scholar 

  18. Murata N, Sato K, Kon J et al (2000) Interaction of sphingosine 1-phosphate with plasma components, including lipoproteins, regulates the lipid receptor-mediated actions. Biochem J 352(Pt 3):809–815

    Article  CAS  PubMed  Google Scholar 

  19. Nofer J-R, Bot M, Brodde M et al (2007) FTY720, a synthetic sphingosine 1 phosphate analogue, inhibits development of atherosclerosis in low-density lipoprotein receptor-deficient mice. Circulation 115:501–508

    Article  CAS  PubMed  Google Scholar 

  20. Lepley D, Paik J-H, Hla T et al (2005) The G protein-coupled receptor S1P2 regulates Rho/Rho kinase pathway to inhibit tumor cell migration. Cancer Res 65:3788–3795

    Article  CAS  PubMed  Google Scholar 

  21. Zhou H, Murthy KS (2004) Distinctive G protein-dependent signaling in smooth muscle by sphingosine 1-phosphate receptors S1P1 and S1P2. Am J Physiol Cell Physiol 286:C1130–C1138

    Article  CAS  PubMed  Google Scholar 

  22. Harvey KA, Paranavitana CN, Zaloga GP et al (2007) Diverse signaling pathways regulate fibroblast differentiation and transformation through Rho kinase activation. J Cell Physiol 211:353–363

    Article  CAS  PubMed  Google Scholar 

  23. Wettschureck N, Offermanns S (2002) Rho/Rho-kinase mediated signaling in physiology and pathophysiology. J Mol Med 80:629–638

    Article  CAS  PubMed  Google Scholar 

  24. Chapados R, Abe K, Ihida-Stansbury K et al (2006) ROCK controls matrix synthesis in vascular smooth muscle cells: coupling vasoconstriction to vascular remodeling. Circ Res 99:837–844

    Article  CAS  PubMed  Google Scholar 

  25. Rivera P, Ocaranza MP, Lavandero S et al (2007) Rho kinase activation and gene expression related to vascular remodeling in normotensive rats with high angiotensin I converting enzyme levels. Hypertension 50:792–798

    Article  CAS  PubMed  Google Scholar 

  26. Oka M, Homma N, Taraseviciene-Stewart L et al (2007) Rho kinase-mediated vasoconstriction is important in severe occlusive pulmonary arterial hypertension in rats. Circ Res 100:923–929

    Article  CAS  PubMed  Google Scholar 

  27. Abe K, Shimokawa H, Morikawa K et al (2004) Long-term treatment with a Rho-kinase inhibitor improves monocrotaline-induced fatal pulmonary hypertension in rats. Circ Res 94:385–393

    Article  CAS  PubMed  Google Scholar 

  28. Tawara S, Fukumoto Y, Shimokawa H (2007) Effects of combined therapy with a Rho-kinase inhibitor and prostacyclin on monocrotaline-induced pulmonary hypertension in rats. J Cardiovasc Pharmacol 50:195–200

    Article  CAS  PubMed  Google Scholar 

  29. Sasaki Y, Suzuki M, Hidaka H (2002) The novel and specific Rho-kinase inhibitor (S)-(+)-2-methyl-1-[(4-methyl-5-isoquinoline)sulfonyl]-homopiperazine as a probing molecule for Rho-kinase-involved pathway. Pharmacol Ther 93:225–232

    Article  CAS  PubMed  Google Scholar 

  30. Hla T, Maciag T (1990) An abundant transcript induced in differentiating human endothelial cells encodes a polypeptide with structural similarities to G-protein-coupled receptors. J Biol Chem 265:9308–9313

    CAS  PubMed  Google Scholar 

  31. MacLennan AJ, Browe CS, Gaskin AA et al (1994) Cloning and characterization of a putative G-protein coupled receptor potentially involved in development. Mol Cell Neurosci 5:201–209

    Article  CAS  PubMed  Google Scholar 

  32. Yamaguchi F, Tokuda M, Hatase O et al (1996) Molecular cloning of the novel human G protein-coupled receptor (GPCR) gene mapped on chromosome 9. Biochem Biophys Res Commun 227:608–614

    Article  CAS  PubMed  Google Scholar 

  33. Graler MH, Bernhardt G, Lipp M (1998) EDG6, a novel G-protein-coupled receptor related to receptors for bioactive lysophospholipids, is specifically expressed in lymphoid tissue. Genomics 53:164–169

    Article  CAS  PubMed  Google Scholar 

  34. Im DS, Heise CE, Ancellin N et al (2000) Characterization of a novel sphingosine 1-phosphate receptor, Edg-8. J Biol Chem 275:14281–14286

    Article  CAS  PubMed  Google Scholar 

  35. Davis MD, Clemens JJ, Macdonald TL et al (2005) Sphingosine 1-phosphate analogs as receptor antagonists. J Biol Chem 280:9833–9841

    Article  CAS  PubMed  Google Scholar 

  36. Sanchez T, Skoura A, Wu MT et al (2007) Induction of vascular permeability by the sphingosine-1-phosphate receptor-2 (S1P2R) and its downstream effectors ROCK and PTEN. Arterioscler Thromb Vasc Biol 27:1312–1318

    Article  CAS  PubMed  Google Scholar 

  37. Riento K, Ridley AJ (2003) Rocks: multifunctional kinases in cell behaviour. Nat Rev Mol Cell Biol 4:446–456

    Article  CAS  PubMed  Google Scholar 

  38. Begum N, Sandu OA, Ito M et al (2002) Active Rho kinase (ROK-alpha) associates with insulin receptor substrate-1 and inhibits insulin signaling in vascular smooth muscle cells. J Biol Chem 277:6214–6222

    Article  CAS  PubMed  Google Scholar 

  39. Lorenz JN, Arend LJ, Robitz R et al (2007) Vascular dysfunction in S1P2 sphingosine 1-phosphate receptor knockout mice. Am J Physiol Regul Integr Compar Physiol 292:R440–R446

    CAS  Google Scholar 

  40. Hu W, Huang J, Mahavadi S et al (2006) Lentiviral siRNA silencing of sphingosine-1-phosphate receptors S1P1 and S1P2 in smooth muscle. Biochem Biophys Res Commun 343:1038–1044

    Article  CAS  PubMed  Google Scholar 

  41. Hu W, Mahavadi S, Huang J et al (2006) Characterization of S1P1 and S1P2 receptor function in smooth muscle by receptor silencing and receptor protection. Am J Physiol Gastrointest Liver Physiol 291:G605–G610

    Article  CAS  PubMed  Google Scholar 

  42. Darland DC, D’Amore PA (2001) TGF beta is required for the formation of capillary-like structures in three-dimensional cocultures of 10T1/2 and endothelial cells. Angiogenesis 4:11–20

    Article  CAS  PubMed  Google Scholar 

  43. Russo S, Bussolati B, Deambrosis I et al (2003) Platelet-activating factor mediates CD40-dependent angiogenesis and endothelial-smooth muscle cell interaction. J Immunol 171:5489–5497

    CAS  PubMed  Google Scholar 

  44. Haberberger RV, Tabeling C, Runciman S et al (2009) Role of sphingosine kinase 1 in allergen-induced pulmonary vascular remodeling and hyperresponsiveness. J Allergy Clin Immunol 124:933–941, e1–9

    Google Scholar 

  45. Lockman K, Hinson JS, Medlin MD et al (2004) Sphingosine 1-phosphate stimulates smooth muscle cell differentiation and proliferation by activating separate serum response factor co-factors. J Biol Chem 279:42422–42430

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors wish to thank Dr. Karen Spear and Charlene Shaffer for their assistance in the preparation of this manuscript and Corine Nivanka Paranavitana for technical assistance.

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Correspondence to Rafat A. Siddiqui.

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Harvey, K.A., Welch, Z., Sliva, D. et al. Role of Rho kinase in sphingosine 1-phosphate-mediated endothelial and smooth muscle cell migration and differentiation. Mol Cell Biochem 342, 7–19 (2010). https://doi.org/10.1007/s11010-010-0461-2

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  • DOI: https://doi.org/10.1007/s11010-010-0461-2

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