A hybrid protein comprising ATF domain of pro-UK and VAS, an angiogenesis inhibitor, is a potent candidate for targeted cancer therapy.
Directional and controllable degradation of extracellular matrix mediated by the uPA-uPA receptor (uPAR) system is ubiquitously implicated in tumor establishment, invasion and metastasis. Targeting the excessive activation of this system as well as the proliferation of the tumor vascular endothelial cell would be expected to prevent tumor neovasculature and halt the tumor development. In this study, we created a fusion protein (ALV), comprising the aminoterminal fragment (ATF) of urokinase and VAS, the antiangiogenic functional domain of vasostatin. The antitumor activity of this hybrid molecule was evaluated with both in vitro and in vivo experiments. Cell adhesion and motility assays demonstrated that ALV, owing to its ATF moiety, could interact with uPAR on the tumor cell surface with high affinity and specificity, and thereby might competitively inhibit the plasmin activation by localized urokinase and contribute to the suppression of tumor invasion. These results and speculations were validated by zymography assay and Matrigel invasion assay. In addition, ALV exhibited an improved inhibitory efficacy against endothelial cell (EC) proliferation and capillary vessel formation in a 3D angiogenesis model, proving that ATF and VAS, when fused into a chimeric molecule, cooperatively inhibited angiogenesis by targeting both the interaction of uPA and uPAR on cell surface (by ATF) and EC proliferation (mainly by VAS). Animal model confirmed that, at the same molar dose, ALV produced significantly higher therapeutic benefit than VAS and ATF in terms of tumor growth delay and mice survival prolongation. Conclusively coupling VAS with the uPAR ligand ATF resulted in an improved antineoplastic activity, which may show a novel avenue for the design of tumor therapeutic drugs.
Sun
Qiming
Q
The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Sciences, Nanjing University, Nanjing 210093, People's Republic of China.
Xu
Qian
Q
Dong
Xiangbai
X
Cao
Lin
L
Huang
Xiaofeng
X
Hu
Qingang
Q
Hua
Zi-Chun
ZC
D000230:Adenocarcinoma / Q000098:blood supply / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D020533:Angiogenesis Inhibitors / Q000235:genetics / Q000494:pharmacology*; D000818:Animals; D001943:Breast Neoplasms / Q000098:blood supply / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D037282:Calreticulin / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D003110:Colonic Neoplasms / Q000098:blood supply / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000098:blood supply / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008819:Mice, Nude; D009369:Neoplasms / Q000098:blood supply* / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000378:metabolism; D055293:Receptors, Urokinase Plasminogen Activator; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D023041:Xenograft Model Antitumor Assays
D020533:Angiogenesis Inhibitors; D037282:Calreticulin; C524898:PLAUR protein, human; D010446:Peptide Fragments; C524899:Plaur protein, mouse; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D011993:Recombinant Fusion Proteins; C121986:vasostatin; D014568:Urokinase-Type Plasminogen Activator