Research Papers:

Targeting the membrane-anchored serine protease testisin with a novel engineered anthrax toxin prodrug to kill tumor cells and reduce tumor burden

Erik W. Martin, Marguerite S. Buzza, Kathryn H. Driesbaugh, Shihui Liu, Yolanda M. Fortenberry, Stephen H. Leppla and Toni M. Antalis _

PDF  |  HTML  |  Supplementary Files  |  How to cite

Oncotarget. 2015; 6:33534-33553. https://doi.org/10.18632/oncotarget.5214

Metrics: PDF 2440 views  |   HTML 2250 views  |   ?  


Erik W. Martin1, Marguerite S. Buzza1, Kathryn H. Driesbaugh1, Shihui Liu2, Yolanda M. Fortenberry3, Stephen H. Leppla2, Toni M. Antalis1

1Center for Vascular and Inflammatory Diseases and the Department of Physiology, University of Maryland School of Medicine, Baltimore, MD 21201, USA

2National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA

3Division of Pediatric Hematology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA

Correspondence to:

Toni M. Antalis, e-mail: [email protected]

Keywords: anthrax toxin, membrane-anchored serine protease, testisin, hepsin, prodrug

Received: July 07, 2015     Accepted: September 03, 2015     Published: September 15, 2015


The membrane-anchored serine proteases are a unique group of trypsin-like serine proteases that are tethered to the cell surface via transmembrane domains or glycosyl-phosphatidylinositol-anchors. Overexpressed in tumors, with pro-tumorigenic properties, they are attractive targets for protease-activated prodrug-like anti-tumor therapies. Here, we sought to engineer anthrax toxin protective antigen (PrAg), which is proteolytically activated on the cell surface by the proprotein convertase furin to instead be activated by tumor cell-expressed membrane-anchored serine proteases to function as a tumoricidal agent. PrAg's native activation sequence was mutated to a sequence derived from protein C inhibitor (PCI) that can be cleaved by membrane-anchored serine proteases, to generate the mutant protein PrAg-PCIS. PrAg-PCIS was resistant to furin cleavage in vitro, yet cytotoxic to multiple human tumor cell lines when combined with FP59, a chimeric anthrax toxin lethal factor-Pseudomonas exotoxin fusion protein. Molecular analyses showed that PrAg-PCIS can be cleaved in vitro by several serine proteases including the membrane-anchored serine protease testisin, and mediates increased killing of testisin-expressing tumor cells. Treatment with PrAg-PCIS also potently attenuated the growth of testisin-expressing xenograft tumors in mice. The data indicates PrAg can be engineered to target tumor cell-expressed membrane-anchored serine proteases to function as a potent tumoricidal agent.

Creative Commons License All site content, except where otherwise noted, is licensed under a Creative Commons Attribution 4.0 License.
PII: 5214