Induction of selective thrombosis and infarction in tumor-feeding vessels represents an attractive strategy to combat cancer. Here we took advantage of the unique coagulation properties of staphylocoagulase and genetically engineered it to generate a new fusion protein with novel anti-cancer properties. This novel bi-functional protein consists of truncated coagulase (tCoa) and an NGR (GNGRAHA) motif that recognizes CD13 and αvβ3 integrin receptors, targeting it to tumor endothelial cells. Herein, we report that tCoa coupled by its C-terminus to an NGR sequence retained its normal binding activity with prothrombin and avβ3 integrins, as confirmed in silico and in vitro. Moreover, in vivo biodistribution studies demonstrated selective accumulation of FITC-labeled tCoa-NGR fusion proteins at the site of subcutaneously implanted PC3 tumor xenografts in nude mice. Notably, systemic administration of tCoa-NGR to mice bearing 4T1 mouse mammary xenografts or PC3 human prostate tumors resulted in a significant reduction in tumor growth. These anti-tumor effects were accompanied by massive thrombotic occlusion of small and large tumor vessels, tumor infarction and tumor cell death. From these findings, we propose tCoa-NGR mediated tumor infarction as a novel and promising anti-cancer strategy targeting both CD13 and integrin αvβ3 positive tumor neovasculature.

Center for Nanobiology and Structural Biology Institute of Microbiology Academy of Sciences of the Czech Republic Zámek 136 373 33 Nové Hrady Czech Republic

Department of Cellular and Molecular Biology Faculty of Biological Science Azarbaijan Shahid Madani University Tabriz Iran

Department of Chemistry University of Neyshabour Khorasan Razavi Province Neyshabour Iran

Department of Clinical Biochemistry and Laboratory Medicine Faculty of Medicine Tabriz University of Medical Sciences Tabriz Iran

Department of Medical Biotechnology Faculty of Advanced Medical Sciences Tabriz University of Medical Sciences Tabriz Iran

Department of Pathobiology Faculty of Veterinary Sciences University of Tabriz Tabriz Iran

Drug Applied Research Center Tabriz University of Medical Sciences Tabriz Iran

Faculty of Science University of South Bohemia Branišovská 1760 37005 České Budějovice Czech Republic

Institute of Animal Breeding and Genetics University of Veterinary Medicine Vienna 1210 Vienna Austria

Iranian National Science Foundation Tehran Iran

Ludwig Boltzmann Institute for Cancer Research 1090 Vienna Austria

Medical University of Vienna 1090 Vienna Austria

North Research Center Pasture Institute of Iran Amol Iran

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