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Utilizing Sphingosine-1-Phosphate to Stimulate Sprouting Angiogenesis

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Sphingosine-1-Phosphate

Part of the book series: Methods in Molecular Biology ((MIMB,volume 874))

Abstract

In vitro models are useful for dissecting cell behavior under controlled conditions. Angiogenesis is a multistep process where endothelial cells (ECs) are activated by pro-angiogenic factors to degrade the basement membrane, migrate into the surrounding matrix, and form sprouting structures connecting neighboring vessels. Sphingosine-1-phosphate (S1P), a biologically active sphingolipid, promotes vessel morphogenesis and angiogenesis during embryonic development and in adults under normal and pathological conditions via its actions on ECs. Here, we describe an in vitro endothelial morphogenic assay that is significantly enhanced by S1P. This method allows for testing whether molecules and their related signaling pathways regulate the initiation of angiogenic sprouts stimulated by S1P, as well as whether individual compounds have pro- or anti-angiogenic properties.

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References

  1. Iruela-Arispe ML, Davis GE (2009) Cellular and molecular mechanisms of vascular lumen formation. Dev Cell 16:222–231

    Article  PubMed  CAS  Google Scholar 

  2. Adams RH, Alitalo K (2007) Molecular regulation of angiogenesis and lymphangiogenesis. Nat Rev Mol Cell Biol 8:464–478

    Article  PubMed  CAS  Google Scholar 

  3. Kalluri R (2003) Basement membranes: structure, assembly and role in tumour angiogenesis. Nat Rev Cancer 3:422–433

    Article  PubMed  CAS  Google Scholar 

  4. Sainson RC, Aoto J, Nakatsu MN, Holderfield M, Conn E, Koller E, Hughes CC (2005) ­Cell-autonomous notch signaling regulates endothelial cell branching and proliferation during vascular tubulogenesis. FASEB J 19:1027–1029

    PubMed  CAS  Google Scholar 

  5. Montesano R, Pepper MS, Vassalli JD, Orci L (1992) Modulation of angiogenesis in vitro. EXS 61:129–136

    PubMed  CAS  Google Scholar 

  6. Nicosia RF, Villaschi S (1999) Autoregulation of angiogenesis by cells of the vessel wall. Int Rev Cytol 185:1–43

    Article  PubMed  CAS  Google Scholar 

  7. Vernon RB, Sage EH (1995) Between molecules and morphology. Extracellular matrix and creation of vascular form. Am J Pathol 147:873–883

    PubMed  CAS  Google Scholar 

  8. Egginton S, Gerritsen M (2003) Lumen formation: in vivo versus in vitro observations. Microcirculation 10:45–61

    PubMed  Google Scholar 

  9. Bayless KJ, Davis GE (2003) Sphingosine-1-phosphate markedly induces matrix metalloproteinase and integrin-dependent human endothelial cell invasion and lumen formation in three-dimensional collagen and fibrin ­matrices. Biochem Biophys Res Commun 312:903–913

    Article  PubMed  CAS  Google Scholar 

  10. Kadish JL, Butterfield CE, Folkman J (1979) The effect of fibrin on cultured vascular endothelial cells. Tissue Cell 11:99–108

    Article  PubMed  CAS  Google Scholar 

  11. Montesano R, Orci L, Vassalli P (1983) In vitro rapid organization of endothelial cells into capillary-like networks is promoted by collagen matrices. J Cell Biol 97:1648–1652

    Article  PubMed  CAS  Google Scholar 

  12. Nicosia RF, Madri JA (1987) The microvascular extracellular matrix. Developmental changes during angiogenesis in the aortic ring-plasma clot model. Am J Pathol 128:78–90

    PubMed  CAS  Google Scholar 

  13. Davis GE, Camarillo CW (1996) An alpha 2 beta 1 integrin-dependent pinocytic mechanism involving intracellular vacuole formation and coalescence regulates capillary lumen and tube formation in three-dimensional collagen matrix. Exp Cell Res 224:39–51

    Article  PubMed  CAS  Google Scholar 

  14. Nakatsu MN, Sainson RC, Aoto JN, Taylor KL, Aitkenhead M, Perez-del-Pulgar S, Carpenter PM, Hughes CC (2003) Angiogenic sprouting and capillary lumen formation ­modeled by human umbilical vein endothelial cells (HUVEC) in fibrin gels: the role of fibroblasts and Angiopoietin-1. Microvasc Res 66:102–112

    Article  PubMed  CAS  Google Scholar 

  15. Liu Y, Senger DR (2004) Matrix-specific ­activation of Src and Rho initiates capillary morphogenesis of endothelial cells. FASEB J 18:457–468

    Article  PubMed  CAS  Google Scholar 

  16. Milstien S, Spiegel S (2006) Targeting sphingosine-1-phosphate: a novel avenue for cancer therapeutics. Cancer Cell 9:148–150

    Article  PubMed  CAS  Google Scholar 

  17. Visentin B, Vekich JA, Sibbald BJ, Cavalli AL, Moreno KM, Matteo RG, Garland WA, Lu Y, Yu S, Hall HS, Kundra V, Mills GB, Sabbadini RA (2006) Validation of an anti-sphingosine-1-phosphate antibody as a potential therapeutic in reducing growth, invasion, and angiogenesis in multiple tumor lineages. Cancer Cell 9:225–238

    Article  PubMed  CAS  Google Scholar 

  18. English D, Brindley DN, Spiegel S, Garcia JG (2002) Lipid mediators of angiogenesis and the signalling pathways they initiate. Biochim Biophys Acta 1582:228–239

    PubMed  CAS  Google Scholar 

  19. Hla T (2004) Physiological and pathological actions of sphingosine-1-phosphate. Semin Cell Dev Biol 15:513–520

    Article  PubMed  CAS  Google Scholar 

  20. Langlois S, Gingras D, Béliveau R (2004) Membrane type 1-matrix metalloproteinase (MT1-MMP) cooperates with sphingosine-1-phosphate to induce endothelial cell migration and morphogenic differentiation. Blood 103:3020–3028

    Article  PubMed  CAS  Google Scholar 

  21. Lee OH, Kim YM, Lee YM, Moon EJ, Lee DJ, Kim JH, Kim KW, Kwon YG (1999) Sphingosine-1-phosphate induces angiogenesis: its angiogenic action and signaling mechanism in human umbilical vein endothelial cells. Biochem Biophys Res Commun 264:743–750

    Article  PubMed  CAS  Google Scholar 

  22. Spiegel S, Milstien S (2003) Sphingosine-1-phosphate: an enigmatic signalling lipid. Nat Rev Mol Cell Biol 4:397–407

    Article  PubMed  CAS  Google Scholar 

  23. 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

    PubMed  CAS  Google Scholar 

  24. Dunlap KA, Erikson DW, Burghardt RC, White FJ, Reed KM, Farmer JL, Spencer TE, Magness RR, Bazer FW, Bayless KJ, Johnson GA (2008) Progesterone and placentation increase secreted phosphoprotein one (SPP1 or osteopontin) in uterine glands and stroma for histotrophic and hematotrophic support of ovine pregnancy. Biol Reprod 79:983–990

    Article  PubMed  CAS  Google Scholar 

  25. Lee MJ, Thangada S, Claffey KP, Ancellin N, Liu CH, Kluk M, Volpi M, Sha’afi RI, Hla T (1999) Vascular endothelial cell adherens junction assembly and morphogenesis induced by sphingosine-1-phosphate. Cell 99:301–312

    Article  PubMed  CAS  Google Scholar 

  26. Endo A, Nagashima K, Kurose H, Mochizuki S, Matsuda M, Mochizuki N (2002) Sphingosine-1-phosphate induces membrane ruffling and increases motility of human umbilical vein endothelial cells via vascular endothelial growth factor receptor and CrkII. J Biol Chem 277:23747–23754

    Article  PubMed  CAS  Google Scholar 

  27. Paik JH, Chae S, Lee MJ, Thangada S, 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 alphavbeta3- and beta1-containing integrins. J Biol Chem 276:11830–11837

    Article  PubMed  CAS  Google Scholar 

  28. Kang H, Bayless KJ, Kaunas R (2008) Fluid shear stress modulates endothelial cell invasion into three-dimensional collagen matrices. Am J Physiol Heart Circ Physiol 295:H2087–H2097

    Article  PubMed  CAS  Google Scholar 

  29. Su SC, Mendoza EA, Kwak HI, Bayless KJ (2008) Molecular profile of endothelial invasion of three-dimensional collagen matrices: insights into angiogenic sprout induction in wound healing. Am J Physiol Cell Physiol 295:C1215–C1229

    Article  PubMed  CAS  Google Scholar 

  30. Argraves KM, Wilkerson BA, Argraves WS, Fleming PA, Obeid LM, Drake CJ (2004) Sphingosine-1-phosphate signaling promotes critical migratory events in vasculogenesis. J Biol Chem 279:50580–50590

    Article  PubMed  CAS  Google Scholar 

  31. Su SC, Maxwell SA, Bayless KJ (2010) Annexin 2 regulates endothelial morphogenesis by controlling AKT activation and junctional integrity. J Biol Chem 285:40624–40634

    Article  PubMed  CAS  Google Scholar 

  32. Kwak HI, Mendoza EA, Bayless KJ (2009) ADAM17 co-purifies with TIMP-3 and modulates endothelial invasion responses in three-dimensional collagen matrices. Matrix Biol 28:470–479

    Article  PubMed  CAS  Google Scholar 

  33. Lee PF, Yeh AT, Bayless KJ (2009) Nonlinear optical microscopy reveals invading endothelial cells anisotropically alter three-dimensional collagen matrices. Exp Cell Res 315:396–410

    Article  PubMed  CAS  Google Scholar 

  34. Maciag T, Cerundolo J, Ilsley S, Kelley PR, Forand R (1979) An endothelial cell growth factor from bovine hypothalamus: identification and partial characterization. Proc Natl Acad Sci USA 76:5674–5678

    Article  PubMed  CAS  Google Scholar 

  35. Bornstein MB (1958) Reconstituted rattail collagen used as substrate for tissue cultures on coverslips in Maximow slides and roller tubes. Lab Invest 7:134–137

    PubMed  CAS  Google Scholar 

  36. Rajan N, Habermehl J, Cote MF, Doillon CJ, Mantovani D (2006) Preparation of ready-to-use, storable and reconstituted type I collagen from rat tail tendon for tissue engineering applications. Nat Protoc 1:2753–2758

    Article  PubMed  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Molecular & Cellular Medicine, Texas A&M Health Science Center, College Station, TX, USA

    Shih-Chi Su & Kayla J. Bayless

Authors
  1. Shih-Chi Su
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  2. Kayla J. Bayless
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Corresponding author

Correspondence to Kayla J. Bayless .

Editor information

Editors and Affiliations

  1. University of Melbourne, Parkville, Victoria, Australia

    Alice Pébay

  2. Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Smyth Road 501, Ottawa, K1Y 8L6, Canada

    Kursad Turksen

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Su, SC., Bayless, K.J. (2012). Utilizing Sphingosine-1-Phosphate to Stimulate Sprouting Angiogenesis. In: Pébay, A., Turksen, K. (eds) Sphingosine-1-Phosphate. Methods in Molecular Biology, vol 874. Humana Press. https://doi.org/10.1007/978-1-61779-800-9_16

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  • DOI: https://doi.org/10.1007/978-1-61779-800-9_16

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-799-6

  • Online ISBN: 978-1-61779-800-9

  • eBook Packages: Springer Protocols

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