A Novel EGFR Targeted Immunotoxin Based on Cetuximab and Type 1 RIP Quinoin Overcomes the Cetuximab Resistance in Colorectal Cancer Cells
Cetuximab is a monoclonal antibody blocking the epidermal growth factor receptor (EGFR) in metastatic colorectal cancer (mCRC). However, cetuximab treatment has no clinical benefits in patients affected by mCRC with KRAS mutation or in the presence of constitutive activation of signalling pathways a...
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MDPI AG
2023-01-01
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| Series: | Toxins |
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| author | Nicola Landi Vincenza Ciaramella Sara Ragucci Angela Chambery Fortunato Ciardiello Paolo V. Pedone Teresa Troiani Antimo Di Maro |
| author_facet | Nicola Landi Vincenza Ciaramella Sara Ragucci Angela Chambery Fortunato Ciardiello Paolo V. Pedone Teresa Troiani Antimo Di Maro |
| author_sort | Nicola Landi |
| collection | DOAJ |
| description | Cetuximab is a monoclonal antibody blocking the epidermal growth factor receptor (EGFR) in metastatic colorectal cancer (mCRC). However, cetuximab treatment has no clinical benefits in patients affected by mCRC with KRAS mutation or in the presence of constitutive activation of signalling pathways acting downstream of the EGFR. The aim of this study was to improve cetuximab’s therapeutic action by conjugating cetuximab with the type 1 ribosome inactivating protein (RIP) quinoin isolated from quinoa seeds. A chemical conjugation strategy based on the use of heterobifunctional reagent succinimidyl 3-(2-pyridyldithio)propionate (SPDP) was applied to obtain the antibody-type 1 RIP chimeric immunoconjugate. The immunotoxin was then purified by chromatographic technique, and its enzymatic action was evaluated compared to quinoin alone. Functional assays were performed to test the cytotoxic action of the quinoin cetuximab immunoconjugate against the cetuximab-resistant GEO-CR cells. The novel quinoin cetuximab immunoconjugate showed a significant dose-dependent cytotoxicity towards GEO-CR cells, achieving IC<sub>50</sub> values of 27.7 nM (~5.0 μg/mL) at 72 h compared to cetuximab (IC<sub>50</sub> = 176.7 nM) or quinoin (IC<sub>50</sub> = 149.3 nM) alone assayed in equimolar amounts. These results support the therapeutic potential of quinoin cetuximab immunoconjugate for the EGFR targeted therapy, providing a promising candidate for further development towards clinical use in the treatment of cetuximab-resistant metastatic colorectal cancer. |
| first_indexed | 2024-03-09T11:06:21Z |
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| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T11:06:21Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-9a09573074fd400b8851133f7ddeb3612023-12-01T00:57:14ZengMDPI AGToxins2072-66512023-01-011515710.3390/toxins15010057A Novel EGFR Targeted Immunotoxin Based on Cetuximab and Type 1 RIP Quinoin Overcomes the Cetuximab Resistance in Colorectal Cancer CellsNicola Landi0Vincenza Ciaramella1Sara Ragucci2Angela Chambery3Fortunato Ciardiello4Paolo V. Pedone5Teresa Troiani6Antimo Di Maro7Department of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, ItalyMedical Oncology, Department of Precision Medicine, University of Campania ‘Luigi Vanvitelli’, Via S. Pansini 5, 80131 Napoli, ItalyDepartment of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, ItalyDepartment of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, ItalyMedical Oncology, Department of Precision Medicine, University of Campania ‘Luigi Vanvitelli’, Via S. Pansini 5, 80131 Napoli, ItalyDepartment of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, ItalyMedical Oncology, Department of Precision Medicine, University of Campania ‘Luigi Vanvitelli’, Via S. Pansini 5, 80131 Napoli, ItalyDepartment of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, ItalyCetuximab is a monoclonal antibody blocking the epidermal growth factor receptor (EGFR) in metastatic colorectal cancer (mCRC). However, cetuximab treatment has no clinical benefits in patients affected by mCRC with KRAS mutation or in the presence of constitutive activation of signalling pathways acting downstream of the EGFR. The aim of this study was to improve cetuximab’s therapeutic action by conjugating cetuximab with the type 1 ribosome inactivating protein (RIP) quinoin isolated from quinoa seeds. A chemical conjugation strategy based on the use of heterobifunctional reagent succinimidyl 3-(2-pyridyldithio)propionate (SPDP) was applied to obtain the antibody-type 1 RIP chimeric immunoconjugate. The immunotoxin was then purified by chromatographic technique, and its enzymatic action was evaluated compared to quinoin alone. Functional assays were performed to test the cytotoxic action of the quinoin cetuximab immunoconjugate against the cetuximab-resistant GEO-CR cells. The novel quinoin cetuximab immunoconjugate showed a significant dose-dependent cytotoxicity towards GEO-CR cells, achieving IC<sub>50</sub> values of 27.7 nM (~5.0 μg/mL) at 72 h compared to cetuximab (IC<sub>50</sub> = 176.7 nM) or quinoin (IC<sub>50</sub> = 149.3 nM) alone assayed in equimolar amounts. These results support the therapeutic potential of quinoin cetuximab immunoconjugate for the EGFR targeted therapy, providing a promising candidate for further development towards clinical use in the treatment of cetuximab-resistant metastatic colorectal cancer.https://www.mdpi.com/2072-6651/15/1/57<i>Chenopodium quinoa</i>chemical conjugationcytotoxicityGEO-CR cellsmonoclonal antibody |
| spellingShingle | Nicola Landi Vincenza Ciaramella Sara Ragucci Angela Chambery Fortunato Ciardiello Paolo V. Pedone Teresa Troiani Antimo Di Maro A Novel EGFR Targeted Immunotoxin Based on Cetuximab and Type 1 RIP Quinoin Overcomes the Cetuximab Resistance in Colorectal Cancer Cells Toxins <i>Chenopodium quinoa</i> chemical conjugation cytotoxicity GEO-CR cells monoclonal antibody |
| title | A Novel EGFR Targeted Immunotoxin Based on Cetuximab and Type 1 RIP Quinoin Overcomes the Cetuximab Resistance in Colorectal Cancer Cells |
| title_full | A Novel EGFR Targeted Immunotoxin Based on Cetuximab and Type 1 RIP Quinoin Overcomes the Cetuximab Resistance in Colorectal Cancer Cells |
| title_fullStr | A Novel EGFR Targeted Immunotoxin Based on Cetuximab and Type 1 RIP Quinoin Overcomes the Cetuximab Resistance in Colorectal Cancer Cells |
| title_full_unstemmed | A Novel EGFR Targeted Immunotoxin Based on Cetuximab and Type 1 RIP Quinoin Overcomes the Cetuximab Resistance in Colorectal Cancer Cells |
| title_short | A Novel EGFR Targeted Immunotoxin Based on Cetuximab and Type 1 RIP Quinoin Overcomes the Cetuximab Resistance in Colorectal Cancer Cells |
| title_sort | novel egfr targeted immunotoxin based on cetuximab and type 1 rip quinoin overcomes the cetuximab resistance in colorectal cancer cells |
| topic | <i>Chenopodium quinoa</i> chemical conjugation cytotoxicity GEO-CR cells monoclonal antibody |
| url | https://www.mdpi.com/2072-6651/15/1/57 |
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