Selective targeted delivery of TNFalpha to tumor blood vessels.
We sought to enhance the selective toxicity of tumor necrosis factor alpha (TNFalpha) to permit its systemic use in cancer therapy. Because ligand-targeted therapeutics have proven successful in improving the selective toxicity of drugs, we prepared a fusion protein (L19mTNFalpha) composed of mouse TNFalpha and a high-affinity antibody fragment (L19 scFv) to the extradomain B (ED-B) domain of fibronectin, a marker of angiogenesis. L19mTNFalpha was expressed in mammalian cells, purified, and characterized. L19mTNFalpha was an immunoreactive and biologically active homotrimer. Radiolabeled L19mTNFalpha selectively targeted tumor neovasculature in tumor-bearing mice, where it accumulated selectively and persistently (tumor-to-blood ratio of the percentage of injected dose per gram [%ID/g] of 700, 48 hours from injection). L19mTNFalpha showed a greater anticancer therapeutic activity than both mTNFalpha and TN11mTNFalpha, a control fusion protein in which an antibody fragment, irrelevant in the tumor model used, substituted for L19. This activity was further dramatically enhanced by its combination with melphalan or the recently reported fusion protein L19-IL2. In conclusion, L19mTNFalpha allows concentrating therapeutically active doses of TNFalpha at the tumor level, thus opening new possibilities for the systemic use of TNFalpha in cancer therapy.
Borsi
Laura
L
Laboratory of Cell Biology, Istituto Nazionale per la Ricerca sul Cancro, Largo Rosanna Benzi, 10 16132 Genoa, Italy. luciano.zardi@tin.it
Balza
Enrica
E
Carnemolla
Barbara
B
Sassi
Francesca
F
Castellani
Patrizia
P
Berndt
Alexander
A
Kosmehl
Hartwig
H
Biro
Attila
A
Siri
Annalisa
A
Orecchia
Paola
P
Grassi
Jessica
J
Neri
Dario
D
Zardi
Luciano
L
D000230:Adenocarcinoma / Q000098:blood supply* / Q000473:pathology; D020533:Angiogenesis Inhibitors / Q000008:administration & dosage* / Q000627:therapeutic use / Q000633:toxicity; D000818:Animals; D000937:Antigen-Antibody Reactions; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use; D003110:Colonic Neoplasms / Q000473:pathology*; D004354:Drug Screening Assays, Antitumor; D004357:Drug Synergism; D005353:Fibronectins / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000008:administration & dosage / Q000235:genetics; D007275:Injections, Intravenous; D007376:Interleukin-2 / Q000008:administration & dosage; D008558:Melphalan / Q000008:administration & dosage; D051379:Mice; D008807:Mice, Inbred BALB C; D009368:Neoplasm Transplantation; D009389:Neovascularization, Pathologic / Q000188:drug therapy*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000627:therapeutic use / Q000633:toxicity; D040521:Subcutaneous Tissue; D018243:Teratocarcinoma / Q000098:blood supply*; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000008:administration & dosage* / Q000235:genetics / Q000627:therapeutic use / Q000633:toxicity
D020533:Angiogenesis Inhibitors; D005353:Fibronectins; D007128:Immunoglobulin Fragments; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D008558:Melphalan