Development and characterization of a potent immunoconjugate targeting the Fn14 receptor on solid tumor cells.

Abstract

TNF-like weak inducer of apoptosis (TWEAK) and fibroblast growth factor (FGF)-inducible 14 (Fn14) are a TNF superfamily ligand-receptor pair involved in many cellular processes including proliferation, migration, differentiation, inflammation, and angiogenesis. The Fn14 receptor is expressed at relatively low levels in normal tissues, but it is known to be dramatically elevated in a number of tumor types, including brain and breast tumors. Thus, it seems to be an excellent candidate for therapeutic intervention. We first analyzed Fn14 expression in human tumor cell lines. Fn14 was expressed in a variety of lines including breast, brain, bladder, skin, lung, ovarian, pancreatic, colon, prostate, and cervical cancer cell lines. We then developed an immunoconjugate containing a high-affinity anti-Fn14 monoclonal antibody (ITEM-4) conjugated to recombinant gelonin (rGel), a highly cytotoxic ribosome-inactivating N-glycosidase. Both ITEM-4 and the conjugate were found to bind to cells to an equivalent extent. Confocal microscopic analysis showed that ITEM4-rGel specifically and rapidly (within 2 hours) internalized into Fn14-positive T-24 bladder cancer cells but not into Fn14-deficient mouse embryonic fibroblasts. Cytotoxicity studies against 22 different tumor cell lines showed that ITEM4-rGel was highly cytotoxic to Fn14-expressing cells and was 8- to 8 × 10(4)-fold more potent than free rGel. ITEM4-rGel was found to kill cells by inducing apoptosis with high-mobility group box 1 protein release. Finally, ITEM4-rGel immunoconjugate administration promoted long-term tumor growth suppression in nude mice bearing T-24 human bladder cancer cell xenografts. Our data support the use of an antibody-drug conjugate approach to selectively target and inhibit the growth of Fn14-expressing tumors.

• Authors: Hong Zhou, John W Marks, Walter N Hittelman, Hideo Yagita, Lawrence H Cheung, Michael G Rosenblum, Jeffrey A Winkles
• Journal: Molecular cancer therapeutics (Mol Cancer Ther) Vol. 10 Issue 7 Pg. 1276-88 (Jul 2011) ISSN: 1538-8514 [Electronic] United States
• PMID: 21586630 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)
• Copyright: © 2011 American Association for Cancer Research.

Chemical References

• Antineoplastic Agents
• HMGB1 Protein
• Immunoconjugates
• Receptors, Tumor Necrosis Factor
• Ribosome Inactivating Proteins, Type 1
• TNFRSF12A protein, human
• TWEAK Receptor
• Tnfrsf12a protein, mouse
• GEL protein, Gelonium multiflorum

Topics

• Animals
• Antineoplastic Agents (isolation & purification, pharmacology)
• Apoptosis (drug effects)
• Cell Line, Tumor
• Female
• Gene Expression Regulation, Neoplastic
• HMGB1 Protein (metabolism)
• HT29 Cells
• Humans
• Immunoconjugates (administration & dosage, isolation & purification, metabolism, pharmacology, toxicity)
• Injections, Intravenous
• Jurkat Cells
• Mice
• Mice, Nude
• Neoplasms (genetics, metabolism, pathology)
• Protein Binding (physiology)
• Receptors, Tumor Necrosis Factor (antagonists & inhibitors, genetics, metabolism)
• Ribosome Inactivating Proteins, Type 1 (metabolism)
• TWEAK Receptor
• Tumor Burden (drug effects)
• Xenograft Model Antitumor Assays