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Targeting the membrane-anchored serine protease testisin with a novel engineered anthrax toxin prodrug to kill tumor cells and reduce tumor burden.
Oncotarget. 2015 Oct 20; 6(32):33534-53.O

Abstract

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.

Authors+Show Affiliations

Center for Vascular and Inflammatory Diseases and the Department of Physiology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.Center for Vascular and Inflammatory Diseases and the Department of Physiology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.Center for Vascular and Inflammatory Diseases and the Department of Physiology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.Division of Pediatric Hematology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.Center for Vascular and Inflammatory Diseases and the Department of Physiology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

26392335

Citation

Martin, Erik W., et al. "Targeting the Membrane-anchored Serine Protease Testisin With a Novel Engineered Anthrax Toxin Prodrug to Kill Tumor Cells and Reduce Tumor Burden." Oncotarget, vol. 6, no. 32, 2015, pp. 33534-53.
Martin EW, Buzza MS, Driesbaugh KH, et al. Targeting the membrane-anchored serine protease testisin with a novel engineered anthrax toxin prodrug to kill tumor cells and reduce tumor burden. Oncotarget. 2015;6(32):33534-53.
Martin, E. W., Buzza, M. S., Driesbaugh, K. H., Liu, S., Fortenberry, Y. M., Leppla, S. H., & Antalis, T. M. (2015). Targeting the membrane-anchored serine protease testisin with a novel engineered anthrax toxin prodrug to kill tumor cells and reduce tumor burden. Oncotarget, 6(32), 33534-53. https://doi.org/10.18632/oncotarget.5214
Martin EW, et al. Targeting the Membrane-anchored Serine Protease Testisin With a Novel Engineered Anthrax Toxin Prodrug to Kill Tumor Cells and Reduce Tumor Burden. Oncotarget. 2015 Oct 20;6(32):33534-53. PubMed PMID: 26392335.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Targeting the membrane-anchored serine protease testisin with a novel engineered anthrax toxin prodrug to kill tumor cells and reduce tumor burden. AU - Martin,Erik W, AU - Buzza,Marguerite S, AU - Driesbaugh,Kathryn H, AU - Liu,Shihui, AU - Fortenberry,Yolanda M, AU - Leppla,Stephen H, AU - Antalis,Toni M, PY - 2015/07/07/received PY - 2015/09/03/accepted PY - 2015/9/23/entrez PY - 2015/9/24/pubmed PY - 2016/9/17/medline KW - anthrax toxin KW - hepsin KW - membrane-anchored serine protease KW - prodrug KW - testisin SP - 33534 EP - 53 JF - Oncotarget JO - Oncotarget VL - 6 IS - 32 N2 - 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. SN - 1949-2553 UR - https://www.unboundmedicine.com/medline/citation/26392335/Targeting_the_membrane_anchored_serine_protease_testisin_with_a_novel_engineered_anthrax_toxin_prodrug_to_kill_tumor_cells_and_reduce_tumor_burden_ L2 - http://www.impactjournals.com/oncotarget/misc/linkedout.php?pii=5214 DB - PRIME DP - Unbound Medicine ER -