Induction of potent TRAIL-mediated tumoricidal activity by hFLEX/Furin/TRAIL recombinant DNA construct.
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been shown to exert selectively cytotoxic activity against many tumor cells but not normal cells. On the other hand, the ligand for the receptor tyrosine kinase Fms-like tyrosine kinase 3 (Flt3L) is a growth factor for hematopoietic progenitors and is a potent stimulating factor for dendritic and NK cells. Previously, we have demonstrated that it is possible to inhibit the outgrowth of primary tumors by the administration of an hFlex (the extracellular domain of the Flt3L) and TRAIL (amino acid residues 95-281) secreted fusion protein. Here, we report that by the insertion of a linker sequence encoding the cleavage site for the Golgi-expressed endoprotease furin between the DNA sequences encoding hFlex and TRAIL, the tumoricidal activity of the cleaved TRAIL protein generated was greatly enhanced in comparison to the hFlex/TRAIL fusion protein. Furthermore, we demonstrate that intratumoral injection of the hFlex/furin/TRAIL DNA, in conjunction with cationic liposomes, significantly suppressed the outgrowth of the human CNE-2 nasopharyngeal tumor xenografts in SCID mice. In situ histological examinations confirmed the expression of TRAIL in the treated tumor nodules and the induction of apoptosis was also evidenced by the presence of numerous pyknotic nuclei.
Wu
Xiaofeng
X
Gene Vector Laboratory, Division of Cellular and Molecular Research, National Cancer Center, 11 Hospital Drive, Singapore 169610, Singapore.
Hui
Kam M
KM
D000818:Animals; D017304:Annexin A5 / Q000032:analysis; D000906:Antibodies / Q000276:immunology; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins; D002277:Carcinoma / Q000473:pathology / Q000628:therapy; D045744:Cell Line, Tumor; D004274:DNA, Recombinant / Q000235:genetics / Q000627:therapeutic use; D003713:Dendritic Cells / Q000276:immunology; D045683:Furin / Q000235:genetics* / Q000627:therapeutic use; D006801:Humans; D007120:Immunochemistry; D008081:Liposomes / Q000627:therapeutic use; D008562:Membrane Glycoproteins / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics* / Q000627:therapeutic use; D051379:Mice; D016513:Mice, SCID; D009303:Nasopharyngeal Neoplasms / Q000473:pathology / Q000628:therapy; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000628:therapy*; D010957:Plasmids / Q000235:genetics / Q000627:therapeutic use; D011419:Propidium / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000627:therapeutic use; D053221:TNF-Related Apoptosis-Inducing Ligand; D014162:Transfection; D016042:Transplantation, Heterotopic; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics*; D023041:Xenograft Model Antitumor Assays
D017304:Annexin A5; D000906:Antibodies; D051017:Apoptosis Regulatory Proteins; D004274:DNA, Recombinant; D008081:Liposomes; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; C505654:Tnfsf10 protein, mouse; D014409:Tumor Necrosis Factor-alpha; C084881:flt3 ligand protein; D011419:Propidium; D045683:Furin