Abstract
IPS-05002, a novel antagonist of integrin a5β1, was screened from a phytochemical compound library using ProteoChip-based integrin a5β1-fibronectin interaction assay method. The novel compound inhibited VEGF-stimulated human umbilical vein endothelial cell (HUVEC) proliferation, adhesion, and migration in a dose-dependent manner. It also suppressed tubular network formation. Differential expression profiling of cell cycle proteins in VEGF-induced HUVECs in the presence of IPS-05002 showed up-regulation of IKB-β, XRCC4, and down-regulation of Cdc6 compared with HUVECs induced by VEGF alone. In conclusion, these data suggest that IPS-05002 will be a potent inhibitor for VEGF-mediated angiogenesis.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Folkman, J. & Shing, Y. Angiogenesis. J. Biol. Chem. 267, 10931–10934 (1991).
Folkman, J. Angiogenesis and apoptosis. Semin. Cancer Biol. 13, 159–167 (2003).
Fan, T.P., Jaggar, R. & Bicknell, R. Controlling the vasculature: angiogenesis, anti-angiogenesis and vascular targeting of gene therapy. Trends Pharmacol. Sci. 16, 57–66 (1995).
Naumov, G.N., Akslen, L.A. & Folkman, J. Role of angiogenesis in human tumor dormancy: animal models of the angiogenic switc. Cell Cycle. 5, 1779–1787 (2006).
Nguyen, M. Angiogenic factors as tumor markers. Invest New Drugs 15, 29–37 (1997).
Rak, J., Yu, J.L., Klement, G. & Kerbel, R.S. Oncogenes and angiogenesis: signaling three-dimensional tumor growth. J. Investig. Dermatol. Symp. Proc. 5, 24–33 (2000).
Avraamides, C.J., Garmy-Susini, B. & Varner, J.A. Integrins in angiogenesis and lymphangiogenesis. Nat. Rev. Cancer 8, 604–617 (2008).
Raymond, K., Faraldo, M.M., Deugnier, M.A. & Glukhova, M.A. Integrins in mammary development. Semin. Cell Dev. Biol. 23, 599–605 (2012).
Barczyk, M., Carracedo, S. & Gullberg, D. Integrins. Cell Tissue Res. 339, 269–280 (2010).
Campbell, I.D. & Humphries, M.J. Integrin structure, activation, and interactions. Cold Spring Harb. Perspect. Biol. 3, a004994 (2011).
Avraamides, C.J., Garmy-Susini, B. & Varner, J.A. Integrins in angiogenesis and lymphangiogenesis. Nat. Rev. Cancer 8, 604–617 (2008).
Carlson, T.R., Hu, H., Braren, R., Kim, Y.H. & Wang, R.A. Cell-autonomous requirement for beta1 integrin in endothelial cell adhesion, migration and survival during angiogenesis in mice. Development 135, 2193–2202 (2008).
Schaffner, F., Ray, A.M. & Dontenwill, M. Integrin a5β1, the Fibronectin Receptor, as a Pertinent Therapeutic Target in Solid Tumors. Cancers 5, 27–47 (2013).
Kim, S., Bell, K., Mousa, S.A. & Varner, J.A. Regulation of angiogenesis in vivo by ligation of integrin alpha5beta1 with the central cell-binding domain of fibronectin. Am. J. Pathol. 156, 1345–1362 (2000).
Magnussen, A. et al. Rapid access of antibodies to alpha5beta1 integrin overexpressed on the luminal surface of tumor blood vessels. Cancer Res. 65, 2712–2721 (2005).
Zhang, H., Li, C. & Baciu, P.C. Expression of integrins and MMPs during alkaline-burn-induced corneal angiogenesis. Invest. Ophthalmol. Vis. Sci. 43, 955–962 (2002).
Bussolati, B., Deambrosis, I., Russo, S., Deregibus, M.C. & Camussi, G. Altered angiogenesis and survival in human tumor-derived endothelial cells. FASEB J. 17, 1159–1161 (2003).
Sudhakar, A. et al. Human tumstatin and human endostatin exhibit distinct antiangiogenic activities mediated by avβ3 and a5β1 integrins. Proc Nat Acad Sci U S A 100, 4766–4771 (2003).
Orecchia, A. et al. Vascular endothelial growth factor receptor-1 is deposited in the extracellular matrix by endothelial cells and is a ligand for the a5β1 integrin. J. Cell. Sci. 116, 3479–3489 (2003).
Felcht, M. et al. Angiopoietin-2 differentially regulates angiogenesis through TIE2 and integrin signaling. J. Clin. Invest. 122, 1991–2005 (2012).
Kim, S., Bakre, M., Yin, H. & Varner, J.A. Inhibition of endothelial cell survival and angiogenesis by protein kinase A. J. Clin. Invest. 110, 933–941 (2002).
Stupack, D., Puente, X.S., Boutsaboualoy, S., Storgard, C.M. & Cheresh, D.A. Apoptosis of adherent cells by recruitment of caspase-8 to unligated integrins. J. Cell Biol. 155, 459–470 (2001).
Bhaskar, V. et al. A function blocking anti-mouse integrin alpha5beta1 antibody inhibits angiogenesis and impedes tumor growth in vivo. J. Transl. Med. 5, 1–11 (2007).
Goodman, S.L. & Picard, M. Integrins as therapeutic targets. Trends Pharmacol. Sci. 33, 405–412 (2012).
Smallheer, J.M. et al. Synthesis and biological evaluation of nonpeptide integrin antagonists containing spirocyclic scaffolds. Bioorg Med Chem Lett. 14, 383–387 (2004).
Bhaskar, V. et al. a chimeric integrin alpha5beta1 antibody, inhibits the growth of VX2 tumors in rabbits. Invest. New Drugs 26, 7–12 (2008).
Livant, D.L. et al. The PHSRN sequence induces extracellular matrix invasion and accelerates wound healing in obese diabetic mice. J. Clin. Invest. 105, 1537–1545 (2000).
Zeng, Z.Z. et al. alpha(5)beta(1) Integrin Ligand PHSRN Induces Invasion and alpha(5) mRNA in Endothelial Cells to Stimulate Angiogenesis. Transl. Oncol. 2, 8–20 (2009).
Kim, E.Y., Bang, J.Y., Chang, S.I. & Kang, I.C. A novel integrin a5β1 antagonistic peptide, A5-1, screened by Protein Chip system as a potent angiogenesis inhibitor. Biochem. Biophys. Res. Commun. 377, 1288–1293 (2008).
Sukhanov, S. & Delafontaine, P. Protein chip-based microarray profiling of oxidized low density lipoprotein-treated cells. Proteomics 5, 1274–1280 (2005).
Koch, S. & Claesson-Welsh, L. Signal transduction by vascular endothelial growth factor receptors. Cold Spring Harb. Perspect. Med. 2, a006502 (2012).
Lee, Y., Kang, D.K., Chang S.I., Han, M.H. & Kang, I.C. High-throughput screening of novel peptide inhibitors of an integrin receptor from the hexapeptide library by using a protein microarray chip. J. Biomol. Screen. 9, 687–694 (2004).
Mechtcheriakova, D. et al. Specificity, diversity, and convergence in VEGF and TNF-a signaling events leading to tissue factor upregulation via EGR-1 in endothelial cells. FASEB J. 15, 230–242 (2001).
Hayden, M.S. & Ghosh, S. Shared principles in NFkappaB signaling. Cell 132, 344–362 (2008).
Hayden, M.S. & Ghosh, S. Regulation of NF-HB by TNF family cytokines. Semin. Immunol. 26, 253–266 (2014).
Herrero, A.B., San Miguel, J. & Gutierrez, N.C. Deregulation of DNA double-strand break repair in multiple myeloma: implications for genome stability. PLoS One 10, e0121581 (2015).
Speck, C., Chen, Z., Li, H. & Stillman, B. ATPase-dependent cooperative binding of ORC and Cdc6 to origin DNA. Nat. Struct. Mol. Biol. 12, 965–971 (2005).
Ahn, E.H. et al. Profiling of Differential Protein Expression in angiogenin-Induced HUVECs using Antibody-Arrayed ProteoChip. Proteomics 6, 1104–1109 (2006).
Ajjappala, B.S. et al. Protein chip analysis of pluripotency-associated proteins in NIH3T3 fibroblast. Proteomics 9, 3968–3978 (2009).
Kang, H., Park, B.R., Yoo, H.S., Kwon, K.R. & Kang, I.C. Anti-angiogenic function of a Korean Ginseng and Toad venom complex, Doksamsumsu-dan (DSSSD) analyzed by a forwarded phase antibody microarray. BioChip Journal 9, 222–231 (2015).
Bang, J.Y. et al. Pharmacoproteomic Analysis of a Novel Cell-permeable Peptide Inhibitor of Tumor-induced Angiogenesis. Mol. Cell. Proteomics 10, 10. 1074/mcp.M110.005264.1-11 (2011).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kim, MA., Kang, IC. Anti-angiogenic mechanism of IPS-05002, a novel antagonist against integrin a5β1, determined by ProteoChip-based antibody array. BioChip J 10, 174–181 (2016). https://doi.org/10.1007/s13206-016-0303-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13206-016-0303-8