Recombinant horseradish peroxidase variants for targeted cancer treatment.

Publisher: John Wiley & Sons Ltd Country of Publication: United States NLM ID: 101595310 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2045-7634 (Electronic) Linking ISSN: 20457634 NLM ISO Abbreviation: Cancer Med Subsets: MEDLINE

Original Publication: [Malden, MA] : John Wiley & Sons Ltd., c2012-

Prodrugs*; Antineoplastic Agents/*pharmacology ; Horseradish Peroxidase/*pharmacology ; Recombinant Proteins/*pharmacology; Antineoplastic Agents/chemistry ; Antineoplastic Agents/isolation & purification ; Cell Line, Tumor ; Cell Survival/drug effects ; Enzyme Activation/drug effects ; Horseradish Peroxidase/chemistry ; Horseradish Peroxidase/isolation & purification ; Humans ; Indoleacetic Acids/chemistry ; Inhibitory Concentration 50 ; Isoenzymes ; Recombinant Proteins/chemistry ; Recombinant Proteins/isolation & purification

Cancer is a major cause of death. Common chemo- and radiation-therapies damage healthy tissue and cause painful side effects. The enzyme horseradish peroxidase (HRP) has been shown to activate the plant hormone indole-3-acetic acid (IAA) to a powerful anticancer agent in in vitro studies, but gene directed enzyme prodrug therapy (GDEPT) studies showed ambivalent results. Thus, HRP/IAA in antibody directed enzyme prodrug therapy (ADEPT) was investigated as an alternative. However, this approach has not been intensively studied, since the enzyme preparation from plant describes an undefined mixture of isoenzymes with a heterogenic glycosylation pattern incompatible with the human system. Here, we describe the recombinant production of the two HRP isoenzymes C1A and A2A in a Pichia pastoris benchmark strain and a glyco-engineered strain with a knockout of the α-1,6-mannosyltransferase (OCH1) responsible for hypermannosylation. We biochemically characterized the enzyme variants, tested them with IAA and applied them on cancer cells. In the absence of H2 O2 , HRP C1A turned out to be highly active with IAA, independent of its surface glycosylation. Subsequent in vitro cytotoxicity studies with human T24 bladder carcinoma and MDA-MB-231 breast carcinoma cells underlined the applicability of recombinant HRP C1A with reduced surface glycoslyation for targeted cancer treatment. Summarizing, this is the first study describing the successful use of recombinantly produced HRP for targeted cancer treatment. Our findings might pave the way for an increased use of the powerful isoenzyme HRP C1A in cancer research in the future.
(© 2016 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)

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P 24861 Austria FWF_ Austrian Science Fund FWF

Keywords: Antibody directed enzyme prodrug therapy (ADEPT); MDA-MB-231 breast carcinoma; Pichia pastoris; T24 bladder carcinoma; horseradish peroxidase; indole-3-acetic acid

0 (Antineoplastic Agents)
0 (Indoleacetic Acids)
0 (Isoenzymes)
0 (Prodrugs)
0 (Recombinant Proteins)
6U1S09C61L (indoleacetic acid)
EC 1.11.1.- (Horseradish Peroxidase)

Date Created: 20160319 Date Completed: 20170118 Latest Revision: 20241211

20250114

PMC4924378

10.1002/cam4.668

26990592