Article thumbnail

Phage-Derived Fully Human Monoclonal Antibody Fragments to Human Vascular Endothelial Growth Factor-C Block Its Interaction with VEGF Receptor-2 and 3

By Matthias Rinderknecht, Alessandra Villa, Kurt Ballmer-Hofer, Dario Neri and Michael Detmar

Abstract

Vascular endothelial growth factor C (VEGF-C) is a key mediator of lymphangiogenesis, acting via its receptors VEGF-R2 and VEGF-R3. High expression of VEGF-C in tumors correlates with increased lymphatic vessel density, lymphatic vessel invasion, sentinel lymph node metastasis and poor prognosis. Recently, we found that in a chemically induced skin carcinoma model, increased VEGF-C drainage from the tumor enhanced lymphangiogenesis in the sentinel lymph node and facilitated metastatic spread of cancer cells via the lymphatics. Hence, interference with the VEGF-C/VEGF-R3 axis holds promise to block metastatic spread, as recently shown by use of a neutralizing anti-VEGF-R3 antibody and a soluble VEGF-R3 (VEGF-C/D trap). By antibody phage-display, we have developed a human monoclonal antibody fragment (single-chain Fragment variable, scFv) that binds with high specificity and affinity to the fully processed mature form of human VEGF-C. The scFv binds to an epitope on VEGF-C that is important for receptor binding, since binding of the scFv to VEGF-C dose-dependently inhibits the binding of VEGF-C to VEGF-R2 and VEGF-R3 as shown by BIAcore and ELISA analyses. Interestingly, the variable heavy domain (VH) of the anti-VEGF-C scFv, which contains a mutation typical for camelid heavy chain-only antibodies, is sufficient for binding VEGF-C. This reduced the size of the potentially VEGF-C-blocking antibody fragment to only 14.6 kDa. Anti-VEGF-C VH-based immunoproteins hold promise to block the lymphangiogenic activity of VEGF-C, which would present a significant advance in inhibiting lymphatic-based metastatic spread of certain cancer types

Topics: Research Article
Publisher: Public Library of Science
OAI identifier: oai:pubmedcentral.nih.gov:2914788
Provided by: PubMed Central

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.

Suggested articles

Citations

  1. (2008). A high-affinity human monoclonal antibody specific to the alternatively spliced EDA domain of fibronectin efficiently targets tumor neo-vasculature in vivo.
  2. (1996). A novel vascular endothelial growth factor, VEGF-C, is a ligand for the Flt4 (VEGFR-3) and KDR (VEGFR-2) receptor tyrosine kinases.
  3. (1998). A recombinant mutant vascular endothelial growth factor-C that has lost vascular endothelial growth factor receptor-2 binding, activation, and vascular permeability activities.
  4. (2009). Alternatively spliced vascular endothelial growth factor receptor-2 is an essential endogenous inhibitor of lymphatic vessel growth.
  5. (2003). Angiogenesis in hematologic malignancies.
  6. (1999). Antibody variable region binding by Staphylococcal protein A: thermodynamic analysis and location of the Fv binding site on E-domain.
  7. (2009). Autocrine loop between vascular endothelial growth factor (VEGF)-C and VEGF receptor-3 positively regulates tumor-associated lymphangiogenesis in oral squamoid cancer cells.
  8. (2001). Betalactamase inhibitors derived from single-domain antibody fragments elicited in the camelidae.
  9. (2009). Complete eradication of human B-cell lymphoma xenografts using rituximab in combination with the immunocytokine L19-IL2.
  10. (2008). Comprehensive analysis of the factors contributing to the stability and solubility of autonomous human VH domains.
  11. (1989). Conformations of immunoglobulin hypervariable regions.
  12. (2000). Design and use of phage display libraries for the selection of antibodies and enzymes.
  13. (2005). Design, construction, and characterization of a large synthetic human antibody phage display library.
  14. (1990). Development of antibodies to unprotected glycosylation sites on recombinant human GM-CSF.
  15. (1993). Evolution of hominoid mitochondrial DNA with special reference to the silent substitution rate over the genome.
  16. (1977). Higher frequencies of transitions among point mutations.
  17. (2005). Inhibition of lymphogenous metastasis using adeno-associated virus-mediated gene transfer of a soluble VEGFR-3 decoy receptor.
  18. (2006). Inhibition of VEGFR-3 activation with the antagonistic antibody more potently suppresses lymph node and distant metastases than inactivation of VEGFR-2.
  19. (2007). Migration-promoting role of VEGF-C and VEGF-C binding receptors in human breast cancer cells.
  20. (2009). Molecular mechanisms of lymph-node metastasis.
  21. (2000). Monoclonal antibodies to vascular endothelial growth factor-D block its interactions with
  22. (1993). Naturally occurring antibodies devoid of light chains.
  23. (1998). Potent enzyme inhibitors derived from dromedary heavy-chain antibodies.
  24. (2001). Recognition of antigens by singledomain antibody fragments: the superfluous luxury of paired domains.
  25. (2002). Role of VEGF family members and receptors in coronary vessel formation.
  26. (2010). Structural determinants of growth factor binding and specificity by VEGF receptor 2.
  27. (2008). Suppression of prostate cancer nodal and systemic metastasis by blockade of the lymphangiogenic axis.
  28. (2002). Suppression of tumor lymphangiogenesis and lymph node metastasis by blocking vascular endothelial growth factor receptor 3 signaling.
  29. (2004). Suppression of VEGFR-3 signaling inhibits lymph node metastasis in gastric cancer.
  30. (2002). The role of tumor lymphangiogenesis in metastatic spread.
  31. (2003). The secretory proprotein convertases furin, PC5, and PC7 activate VEGF-C to induce tumorigenesis.
  32. (1998). The structure of human interferon-beta: implications for activity.
  33. (2008). Tumor lymphangiogenesis and melanoma metastasis.
  34. (2003). Tumor lymphangiogenesis: a novel prognostic indicator for cutaneous melanoma metastasis and survival.
  35. (2006). Tumor-targeting properties of novel antibodies specific to the large isoform of tenascin-C.
  36. (1999). Unique single-domain antigen binding fragments derived from naturally occurring camel heavy-chain antibodies.
  37. (1999). Vascular endothelial growth factor (VEGF) and its receptors in tumor-bearing dogs.
  38. (2002). Vascular endothelial growth factor (VEGF)-C signaling through FLT-4 (VEGFR-3) mediates leukemic cell proliferation, survival, and resistance to chemotherapy.
  39. (2006). Vascular endothelial growth factor (VEGF)/VEGF-C mosaic molecules reveal specificity determinants and feature novel receptor binding patterns.
  40. (2008). Vascular endothelial growth factor C stimulates progression of human gastric cancer via both autocrine and paracrine mechanisms.
  41. (1998). Vascular endothelial growth factor D (VEGF-D) is a ligand for the tyrosine kinases
  42. (2006). Vascular endothelial growth factor-D activates VEGFR-3 expressed in osteoblasts inducing their differentiation.
  43. (2002). VEGF-C mediates cyclic pressure-induced endothelial cell proliferation.
  44. (2008). VEGF-C, a lymphatic growth factor, is a RANKL target gene in osteoclasts that enhances osteoclastic bone resorption through an autocrine mechanism.
  45. (2001). VEGF-D promotes the metastatic spread of tumor cells via the lymphatics.