Potential of unglycosylated horseradish peroxidase variants for enzyme prodrug cancer therapy
| Autor: | Oliver Spadiut, Angelika Wlodarczyk, Mario Kuttke, Christina Divne, Valentina Furlanetto, Diana Humer, Anna-Katharina Schruef |
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| Rok vydání: | 2021 |
| Předmět: |
Cell Survival
Antineoplastic Agents RM1-950 Adenocarcinoma Horseradish peroxidase Enzyme engineering Mice In vivo Cell Line Tumor medicine Escherichia coli Cytotoxic T cell Animals Humans Prodrugs Viability assay Cytotoxicity Horseradish Peroxidase Pharmacology biology Toxicity Indoleacetic Acids Chemistry Cancer food and beverages Nasopharyngeal Neoplasms General Medicine Prodrug medicine.disease HCT116 Cells Mice Inbred C57BL Enzyme prodrug cancer therapy Recombinant enzyme Cancer cell Colonic Neoplasms biology.protein Cancer research Carcinoma Squamous Cell Female Therapeutics. Pharmacology |
| Zdroj: | Biomedicine & Pharmacotherapy, Vol 142, Iss, Pp 112037-(2021) |
| ISSN: | 1950-6007 |
| Popis: | Fighting cancer still relies on chemo- and radiation therapy, which is a trade-off between effective clearance of malignant cells and severe side effects on healthy tissue. Targeted cancer treatment on the other hand is a promising and refined strategy with less systemic interference. The enzyme horseradish peroxidase (HRP) exhibits cytotoxic effects on cancer cells in combination with indole-3-acetic acid (IAA). However, the plant-derived enzyme is out of bounds for medical purposes due to its foreign glycosylation pattern and resulting rapid clearance and immunogenicity. In this study, we generated recombinant, unglycosylated HRP variants in Escherichia coli using random mutagenesis and investigated their biochemical properties and suitability for cancer treatment. The cytotoxicity of the HRP-IAA enzyme prodrug system was assessed in vitro with HCT-116 human colon, FaDu human nasopharyngeal squamous cell carcinoma and murine colon adenocarcinoma cells (MC38). Extensive cytotoxicity was shown in all three cancer cell lines: the cell viability of HCT-116 and MC38 cells treated with HRP-IAA was below 1% after 24 h incubation and the surviving fraction of FaDu cells was ≤ 10% after 72 h. However, no cytotoxic effect was observed upon in vivo intratumoral application of HRP-IAA on a MC38 tumor model in C57BL/6J mice. However, we expect that targeting of HRP to the tumor by conjugation to specific antibodies or antibody fragments will reduce HRP clearance and thereby enhance therapy efficacy. |
| Databáze: | OpenAIRE |
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