Activation of CD95L fusion protein prodrugs by tumor-associated proteases.
To achieve tumor cell-restricted activation of CD95, we developed a CD95L fusion protein format, in which CD95L activity is only unmasked upon antibody-mediated binding to tumor cells and subsequent processing by tumor-associated proteases, such as matrix metalloproteases (MMPs) and urokinase plasminogen activator (uPA). On target-negative, but MMP- and uPA-expressing HT1080 tumor cells, the CD95L prodrugs were virtually inactive. On target antigen-expressing HT1080 cells, however, the CD95L prodrugs showed an apoptotic activity comparable to soluble CD95L artificially activated by crosslinking. CD95 activation by the CD95L prodrugs was preceded by prodrug processing. Apoptosis was blocked by inhibitors of MMPs or uPA and by neutralizing antibodies recognizing the targeted cell surface antigen or the CD95L moiety of the prodrugs. In a xenotransplantation tumor model, local application of the prodrug reduced the growth of target antigen-expressing, but not antigen-negative tumor cells, verifying targeted CD95L prodrug activation in vivo.
Watermann
I
I
Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, Germany.
Gerspach
J
J
Lehne
M
M
Seufert
J
J
Schneider
B
B
Pfizenmaier
K
K
Wajant
H
H
D000818:Animals; D000951:Antigens, Neoplasm / Q000378:metabolism; D000970:Antineoplastic Agents / Q000378:metabolism*; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D014408:Biomarkers, Tumor / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D053222:Fas Ligand Protein / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D005354:Fibrosarcoma / Q000188:drug therapy / Q000378:metabolism; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D011355:Prodrugs / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D011994:Recombinant Proteins; D036401:Toxicity Tests, Chronic; D014407:Tumor Cells, Cultured; D019014:fas Receptor / Q000187:drug effects / Q000378:metabolism*
D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D053222:Fas Ligand Protein; D011355:Prodrugs; D011994:Recombinant Proteins; D019014:fas Receptor; D020778:Matrix Metalloproteinase 2