IMMUNE NETWORK

The Korean Association of Immunobiologists (대한면역학회)
권호 표지

Development of Evaluating Ways for the Efficacy of Anti-VEGF Biopharmaceuticals

VEGF 제어의약품의 효능 평가법 개발

  • Nam, Eun-Hee (Department of Molecular Bioscience, School of Bioscience & Biotechnology, Kangwon National University) ;
  • Jeon, Seong-Hyun (Department of Molecular Bioscience, School of Bioscience & Biotechnology, Kangwon National University) ;
  • Lee, Wha-Jung (Department of Molecular Bioscience, School of Bioscience & Biotechnology, Kangwon National University) ;
  • Seo, Dong-Wan (Department of Molecular Bioscience, School of Bioscience & Biotechnology, Kangwon National University) ;
  • Kim, Pyeung-Hyeun (Department of Molecular Bioscience, School of Bioscience & Biotechnology, Kangwon National University)
  • 남은희 (강원대학교 BT특성화대학 분자생명과학과) ;
  • 전성현 (강원대학교 BT특성화대학 분자생명과학과) ;
  • 이화정 (강원대학교 BT특성화대학 분자생명과학과) ;
  • 서동완 (강원대학교 BT특성화대학 분자생명과학과) ;
  • 김평현 (강원대학교 BT특성화대학 분자생명과학과)
  • Published : 2007.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

Background: Angiogenesis mediated by VEGF constitutes a new target for anti-cancer therapy which has explored through different ways of intervention aiming at the blocking of the tumoral angiogenesis. In the present study, we developed the assays by which efficacies of anti-VEGF inhibitor candidates are evaluated at the various levels. Methods & Results: First, we developed two sandwich ELISAs using coated anti-VEGF Ab and soluble Flt-1 receptor fusion protein (sFlt-1/Fc). As low as 200 pg/ml of hVEGF diluted in human sera was detectable by these assays. In addition, we found that VEGF inhibitors ($2{\mu}g/ml$ of either anti-VEGF Ab or sFlt-1/Fc) completely block 5 ng/ml VEGF in these ELISAs. Subsequently, two bioassays, wound healing and HUVEC tube formation assays, revealed that anti-VEGF Ab $(1{\mu}g/ml)$ & sFlt-1/Fc Ab $(1{\mu}g/ml)$, or SU5416 (VEGFR tyrosine kinase inhibitor, $1{\mu}M$) prevents the activity of VEGF $(1{\sim}10ng/ml)$. Finally, secretion of MMP-9 by VEGF-stimulated macrophages was abolished by treatment of anti-VEGF Ab $(1{\mu}g/ml)$ in gelatin zymography. Conclusion: ELISAs together with bioassays developed in this study are appropriate for evaluation of the efficacy of inhibitors of VEGF.

Keywords

References

  1. Ferrara N: VEGF and the quest for tumour angiogenesis factors. Nat Rev Cancer 2;795-803, 2002 https://doi.org/10.1038/nrc909
  2. Jain RK: Molecular regulation of vessel maturation. Nat Med 9;685-693, 2003 https://doi.org/10.1038/nm0603-685
  3. Gille H, Kowalski J, Li B, LeCouter J, Moffat B, Zioncheck TF, Pelletier N, Ferrara N: Analysis of biological effects and signaling properties of Flt-1 (VEGFR-1) and KDR (VEGFR-2). A reassessment using novel receptor-specific vascular endothelial growth factor mutants. J Biol Chem 276;3222-3230, 2001 https://doi.org/10.1074/jbc.M002016200
  4. Meyer M, Clauss M, Lepple-Wienhues A, Waltenberger J, Augustin HG, Ziche M, Lanz C, Buttner M, Rziha HJ, Dehio C: A novel vascular endothelial growth factor encoded by Orf virus, VEGF-E, mediates angiogenesis via signalling through VEGFR-2 (KDR) but not VEGFR-1 (Flt-1) receptor tyrosine kinases. Embo J 18;363-374, 1999 https://doi.org/10.1093/emboj/18.2.363
  5. Hurwitz H, Fehrenbacher L, Novotny W, Cartwright T, Hainsworth J, Heim W, Berlin J, Baron A, Griffing S, Holmgren E, Ferrara N, Fyfe G, Rogers B, Ross R, Kabbinavar F: Bevacizumab plus irinotecan, fluorouracil, and leucovorin for metastatic colorectal cancer. N Engl J Med 350;2335-2342, 2004 https://doi.org/10.1056/NEJMoa032691
  6. Johnson DH, Fehrenbacher L, Novotny WF, Herbst RS, Nemunaitis JJ, Jablons DM, Langer CJ, DeVore RF, Gaudreault 3rd J, Damico LA, Holmgren E, Kabbinavar F: Randomized phase II trial comparing bevacizumab plus carboplatin and paclitaxel with carboplatin and paclitaxel alone in previously untreated locally advanced or metastatic non-small- cell lung cancer. J Clin Oncol 22;2184-2191, 2004 https://doi.org/10.1200/JCO.2004.11.022
  7. Motzer RJ, Michaelson MD, Redman BG, Hudes GR, Wilding G, Figlin RA, Ginsberg MS, Kim ST, Baum CM, DePrimo SE, Li JZ, Bello CL, Theuer CP, George DJ, Rini BI: Activity of SU11248, a multitargeted inhibitor of vascular endothelial growth factor receptor and platelet-derived growth factor receptor, in patients with metastatic renal cell carcinoma. J Clin Oncol 24;16-24, 2006 https://doi.org/10.1200/JCO.2005.02.2574
  8. Escudier B, Eisen T, Stadler WM, Szczylik C, Oudard S, Siebels M, Negrier S, Chevreau C, Solska E, Desai AA, Rolland F, Demkow T, Hutson TE, Gore M, Freeman S, Schwartz B, Shan M, Simantov R, Bukowski RM: Sorafenib in advanced clear-cell renal-cell carcinoma. N Engl J Med 356;125-134, 2007 https://doi.org/10.1056/NEJMoa060655
  9. Drevs J, Siegert P, Medinger M, Mross K, Strecker R, Zirrgiebel U, Harder J, Blum H, Robertson J, Jurgensmeier JM, Puchalski TA, Young H, Saunders O, Unger C: Phase I clinical study of AZD2171, an oral vascular endothelial growth factor signaling inhibitor, in patients with advanced solid tumors. J Clin Oncol 25;3045-3054, 2007 https://doi.org/10.1200/JCO.2006.07.2066
  10. Ogawa T, Takayama K, Takakura N, Kitano S, Ueno H: Anti-tumor angiogenesis therapy using soluble receptors: enhanced inhibition of tumor growth when soluble fibroblast growth factor receptor-1 is used with soluble vascular endothelial growth factor receptor. Cancer Gene Ther 9;633-640, 2002 https://doi.org/10.1038/sj.cgt.7700478
  11. Jaffe EA, Nachman RL, Becker CG, Minick CR: Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 52;2745-2756, 1973 https://doi.org/10.1172/JCI107470
  12. Jeon SH, Chae BC, Kim HA, Seo GY, Seo DW, Chun GT, Kim NS, Yie SW, Byeon WH, Eom SH, Ha KS, Kim YM, PH Kim: Mechanisms underlying TGF-{beta}1-induced expression of VEGF and Flk-1 in mouse macrophages and their implications for angiogenesis. J Leukoc Biol 81;557-566, 2007 https://doi.org/10.1189/jlb.0806517
  13. Roy H, Bhardwaj S, Yla-Herttuala S: Biology of vascular endothelial growth factors. FEBS Lett 580;2879-2887, 2006 https://doi.org/10.1016/j.febslet.2006.03.087
  14. Jeon SH, Chae BC, Kim HA, Seo GY, Seo DW, Chun GT, Yie SW, Eom SH, Kim PH: The PKA/CREB pathway is closely involved in VEGF expression in mouse macrophages. Mol Cells 23;23-29, 2007
  15. Bergers G, Brekken R, McMahon G, Vu TH, Itoh T, Tamaki K, Tanzawa K, Thorpe P, Itohara S, Werb Z, Hanahan D: Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis. Nat Cell Biol 2;737-744, 2000 https://doi.org/10.1038/35036374