Effects of a novel cyclic RGD peptidomimetic on cell proliferation, migration and angiogenic activity in human endothelial cells
Tóm tắt
Cyclic RGD peptidomimetics containing a bifunctional diketopiperazine scaffold are a novel class of high-affinity ligands for the integrins αVβ3 and αVβ5. Since integrins are a promising target for the modulation of normal and pathological angiogenesis, the present study aimed at characterizing the ability of the RGD peptidomimetic cyclo[DKP-RGD] 1 proliferation, migration and network formation in human umbilical vein endothelial cells (HUVEC). Cell viability was assessed by flow cytometry and annexin V (ANX)/propidium iodide (PI) staining. Cell proliferation was evaluated by the ELISA measurement of bromodeoxyuridine (BrdU) incorporation. Network formation by HUVEC cultured in Matrigel-coated plates was evaluated by optical microscopy and image analysis. Integrin subunit mRNA expression was assessed by real time-PCR and Akt phosphorylation by western blot analysis. Cyclo[DKP-RGD] 1 does not affect cell viability and proliferation either in resting conditions or in the presence of the pro-angiogenic growth factors VEGF, EGF, FGF, and IGF-I. Addition of cyclo[DKP-RGD] 1 however significantly decreased network formation induced by pro-angiogenic growth factors or by IL-8. Cyclo[DKP-RGD] 1 did not affect mRNA levels of αV, β3 or β5 integrin subunits, however it significantly reduced the phosphorylation of Akt. Cyclo[DKP-RGD] 1 can be a potential modulator of angiogenesis induced by different growth factors, possibly devoid of the adverse effects of cytotoxic RGD peptidomimetic analogues.
Tài liệu tham khảo
Carmeliet P: Mechanisms of angiogenesis and arteriogenesis. Nat Med. 2000, 6: 389-395. 10.1038/74651.
D’Andrea LD, Del Gatto A, Pedone C, Benedetti E: Peptide-based molecules in angiogenesis. Chem Biol Drug Des. 2006, 67: 115-126. 10.1111/j.1747-0285.2006.00356.x.
Li A, Dubey S, Varney ML, Bhavana J: IL-8 Directly enhanced endothelial cell survival, proliferation, and matrix metalloproteinases production and regulated angiogenesis. J Immunol. 2003, 170: 3369-3376. 10.4049/jimmunol.170.6.3369.
Juliano RL: Signal transduction by cell adhesion receptor and the cytoskeleton: function of integrins, cadherins, selectins, and immunoglobulin-superfamily members. Annu Rev Pharmacol Toxicol. 2002, 42: 283-323. 10.1146/annurev.pharmtox.42.090401.151133.
Hodivala-Dilke KM, Reynolds AR, Reynolds LE: Integrins in angiogenesis: multitalented molecules in a balancing act. Cell Tissue Res. 2003, 314: 131-144. 10.1007/s00441-003-0774-5.
Plow EF, Haas TA, Zhang L, Loftus J, Smith JW: Ligand binding to integrins. J Biol Chem. 2000, 275: 21785-21788. 10.1074/jbc.R000003200.
Xiong JP, Stehle T, Zhang R, Joachimiak A, Frech M, Goodman SL, Arnaout MA: Crystal structure of the extracellular segment of integrin alpha V beta 3 in complex with an Arg-Gly-Asp ligand. Science. 2002, 296: 151-155. 10.1126/science.1069040.
Paolillo M, Russo MA, Serra M, Colombo L, Schinelli S: Small molecule integrin antagonists in cancer therapy. Mini-Rev Med Chem. 2009, 9: 1439-1446. 10.2174/138955709789957404.
Schottelius M, Laufer B, Kessler H, Wester HJ: Ligands for mapping alphavbeta3-integrin expression in vivo. Acc Chem Res. 2009, 42 (7): 969-980. 10.1021/ar800243b.
Mas-Moruno C, Rechenmacher F, Kessler H: Cilengitide: the first anti-angiogenic small molecule drug candidate. Design, synthesis and clinical evaluation. Anti-Cancer Agents Med Chem. 2010, 10: 753-768. 10.2174/187152010794728639.
Kim YH, Lee JK, Kim B, DeWitt JP, Lee JE, Han JH, Kim SK, Oh CW, Kim CY: Combination therapy of cilengitide with belotecan against experimental glioblastoma. Int J Cancer. 2013, 133: 749-756. 10.1002/ijc.28058.
Wan W, Guo N, Pan D, Yu C, Weng Y, Luo S, Ding H, Xu Y, Wang L, Lang L, Xie Q, Yang M, Chen X: First experience of 18 F-alfatide in lung cancer patients using a new lyophilized kit for rapid radiofluorination. J Nucl Med. 2013, 54: 691-698. 10.2967/jnumed.112.113563.
Reynolds AR, Hart IR, Watson AR, Welti JC, Silva RG, Robinson SD, Da Violante G, Gourlaouen M, Salih M, Jones MC, Jones DT, Saunders G, Kostourou V, Perron-Sierra F,JC, Tucker GC, Hodivala-Dilke KM: Stimulation of tumor growth and angiogenesis by low concentrations of RGD-mimetic integrin inhibitors. Nat Med. 2009, 15: 392-400. 10.1038/nm.1941.
Papo N, Silverman AP, Lahti JL, Cochran JR: Antagonistic VEGF variants engineered to simultaneously bind to and inhibit VEGFR2 and αvβ3 integrin. Proc Natl Acad Sci U S A. 2011, 108: 14067-14072. 10.1073/pnas.1016635108.
Ressurreiçao ASM, Vidu A, Civera M, Belvisi L, Potenza D, Manzoni L, Ongeri S, Gennari C, Piarulli U: Cyclic RGD-peptidomimetics containing bifunctional diketopiperazine scaffolds as new potent integrin ligands. Chem Eur J. 2009, 15: 12184-12188. 10.1002/chem.200902398.
Marchini M, Mingozzi M, Colombo R, Guzzetti I, Belvisi L, Vasile F, Potenza D, Piarulli U, Arosio D, Gennari C: Cyclic RGD-peptidomimetics containing bifunctional diketopiperazine scaffolds as new potent integrin ligands. Chem Eur J. 2012, 18: 6195-6207. 10.1002/chem.201200457.
Shiojima I, Walsh K: Role of Akt signaling in vascular homeostasis and angiogenesis. Circ Res. 2002, 90: 1243-1250. 10.1161/01.RES.0000022200.71892.9F.
Russo MA, Paolillo M, Sanchez-Hernandes Y, Curti D, Ciusani E, Serra M, Colombo L, Schinelli S: A small-molecule RDG-integrin antagonist inhibits cell adhesion, cell migration and induces anoikis in glioblastoma cells. International J of Oncology. 2013, 42: 83-92.
Xu Y, Zhou Y, Lin H, Hu H, Wang Y, Xu G: Toll-like receptor 2 in promoting angiogenesis after acute ischemic injury. Int J Mol Med Mar. 2013, 31: 555-560.
Hynes RO: Cell-matrix adhesion in vascular development. J Thromb Haemost. 2007, 1: 32-40.
Kumar CC: Integrin alpha v beta 3 as a therapeutic target for blocking tumor-induced angiogenesis. Curr Drug Targets. 2003, 4: 123-131. 10.2174/1389450033346830.
Baranska P, Jerczynska H, Pawlowska Z, Koziolkiewicz W, Cierniewski CS: Expression of integrins and adhesive properties of human endothelial cell line EA.hy 926. Cancer Genomics Proteomics. 2005, 2: 265-270.
Lomonaco SL, Finniss S, Xiang CL, Lee HK, Jiang W, Lemke N, Rempel SA, Mikkelsen T, Brodie C: Cilengitide induces autophagy-mediated cell death in glioma cells. Neuro Oncol. 2011, 13: 857-865. 10.1093/neuonc/nor073.
Moreno PR, Purushothaman M, Purushothaman KR: Plaque neovascularization: defense mechanisms, betrayal, or a war in progress. Ann NY Acad Sci. 2012, 1254: 7-17. 10.1111/j.1749-6632.2012.06497.x.
Colombo R, Mingozzi M, Belvisi L, Arosio D, Piarulli U, Carenini N, Perego P, Zaffaroni N, De Cesare M, Castiglioni V, Scanziani E, Gennari C: Synthesis and biological evaluation (in vitro and in vivo) of cyclic arginine-glycine-aspartate (RGD) peptidomimetic-paclitaxel conjugates targeting integrin αVβ3. J Med Chem. 2012, 55: 10460-10474. 10.1021/jm301058f.
Eliceiri BP: Integrin and growth factor receptor crosstalk. Circ Res. 2001, 89: 1104-1110. 10.1161/hh2401.101084.
Shabbir SH, Eisenberg JL, Mrksich M: An Inhibitor of a Cell Adhesion Receptor Stimulates Cell Migration. Angew Chem Int Ed. 2010, 49: 7706-7709. 10.1002/anie.201002699.
Waugh DJJ, Wilson C: The interleukin-8 pathway in cancer. Clin Cancer Res. 2008, 14: 6735-6741. 10.1158/1078-0432.CCR-07-4843.
Brooks AN, Kilgour E, Smith PD: Molecular pathways: fibroblast growth factor signaling: a new therapeutic opportunity in cancer. Clin Cancer Res. 2012, 18: 1855-1862. 10.1158/1078-0432.CCR-11-0699.
Paesler J, Gehrke I, Poll-Wolbeck SJ, Kreuzer KA: Targeting the vascular endothelial growth factor in hematologic malignancies. Eur J Haematol. 2012, 89: 373-384. 10.1111/ejh.12009.
Hu B, Wei YQ, Tian L, Zhao X, Lu Y, Wu Y, Yao B, Liu JY, Niu T, Wen YJ, He QM, Su JM, Huang MJ, Lou YY, Luo Y, Kan B: Active antitumor immunity elicited by vaccine based on recombinant form of epidermal growth factor receptor. J Immunother. 2005, 28: 236-244. 10.1097/01.cji.0000161394.11831.3f.
Moulton KS, Karen S: Angiogenesis in atherosclerosis: gathering evidence beyond speculation. Curr Opin Lipidol. 2006, 17: 548-555. 10.1097/01.mol.0000245261.71129.f0.