In vitro and in vivo effects of an anti-mouse endoglin (CD105)-immunotoxin on the early stages of mouse B16MEL4A5 melanoma tumours.
TGF-beta superfamily co-receptors are emerging as targets for cancer therapy, acting both directly on cells and indirectly on the tumour neovasculature. Endoglin (CD105), an accessory component of the TGF-beta receptor complex, is expressed in certain melanoma cell lines and the endothelial cells of tumour neovessels. Targeting endoglin with immunotoxins is an attractive approach for actively suppressing the blood supply to tumours. Here, we report evidence indicating that endoglin is expressed in mouse melanoma B16MEL4A5 and mouse fibroblast L929 cell lines. We prepared an immunotoxin to target endoglin by coupling the rat anti-mouse MJ7/18 (IgG2a) monoclonal antibody (mAb) to the non-toxic type 2 ribosome-inactivating protein nigrin b (Ngb) with N-succinimidyl 3-(2-pyridyldithio)-propionate (SPDP) as a linker with a molar nigrin b at a MJ7/18 stoichiometry of 2:1. The MJ7-Ngb immunotoxin generated killed both cell lines, with IC50 values of 4.2 × 10(-9) M for B16MEL4A5 and 7.7 × 10(-11) M for L929 cells. For in vivo assays of the immunotoxin, B16MEL4A5 cells were injected subcutaneously into the right flanks of 6-week-old C57BL/6 J mice. When the animals developed palpable solid tumours, they were subjected to treatment with the immunotoxin. While treatment with either MJ7/18 mAb or Ngb did not affect tumour development, treatment with the immunotoxin completely and steadily blocked tumour growth up to 7 days, after which some tumours re-grew. Thus, vascular-targeting therapy with this anti-vascular immunotoxin could promote the destruction of newly created tumour vessels at early stages of B16MEL4A5 tumour development and readily accessible CD105+ B16MEL4A5 melanoma cells.
Facultad de Ciencias, Departamento de Bioquímica y Biología Molecular, Universidad de Valladolid, 47005, Valladolid, Spain.
Raquel
R
Muñoz
Yolanda
Y
Arias
José Miguel
JM
Ferreras
Pilar
P
Jiménez
Carmen
C
Langa
María Angeles
MA
Rojo
Manuel José
MJ
Gayoso
Damián
D
Córdoba-Díaz
Carmelo
C
Bernabéu
Tomás
T
Girbés
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology / Q000627:therapeutic use*; D015703:Antigens, CD / Q000276:immunology*; D002460:Cell Line; D045744:Cell Line, Tumor; D000071063:Endoglin; D000922:Immunotoxins / Q000494:pharmacology / Q000627:therapeutic use*; D008546:Melanoma, Experimental / Q000098:blood supply / Q000628:therapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D009389:Neovascularization, Pathologic / Q000188:drug therapy; D010940:Plant Proteins / Q000008:administration & dosage*; D011956:Receptors, Cell Surface / Q000276:immunology*; D054788:Ribosome Inactivating Proteins / Q000008:administration & dosage*
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; C064206:ENG protein, human; D000071063:Endoglin; D000922:Immunotoxins; D010940:Plant Proteins; D011956:Receptors, Cell Surface; C579416:nigrin b protein, Sambucus nigra; D054788:Ribosome Inactivating Proteins