The Significance of Cell Surface N-Glycosylation for Internalization and Potency of Cytotoxic Conjugates Targeting Receptor Tyrosine Kinases
Precise anticancer therapies employing cytotoxic conjugates constitute a side-effect-limited, highly attractive alternative to commonly used cancer treatment modalities, such as conventional chemotherapy, radiotherapy or surgical interventions. Receptor tyrosine kinases are a large family of N-glyco...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-07-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/23/15/8514 |
| _version_ | 1797432836545839104 |
|---|---|
| author | Marta Poźniak Dominika Żukowska Aleksandra Gędaj Mateusz Adam Krzyścik Natalia Porębska Małgorzata Zakrzewska Jacek Otlewski Łukasz Opaliński |
| author_facet | Marta Poźniak Dominika Żukowska Aleksandra Gędaj Mateusz Adam Krzyścik Natalia Porębska Małgorzata Zakrzewska Jacek Otlewski Łukasz Opaliński |
| author_sort | Marta Poźniak |
| collection | DOAJ |
| description | Precise anticancer therapies employing cytotoxic conjugates constitute a side-effect-limited, highly attractive alternative to commonly used cancer treatment modalities, such as conventional chemotherapy, radiotherapy or surgical interventions. Receptor tyrosine kinases are a large family of N-glycoproteins intensively studied as molecular targets for cytotoxic conjugates in various cancers. At the cell surface, these receptors are embedded in a dense carbohydrate layer formed by numerous plasma membrane glycoproteins. The complexity of the cell surface architecture is further increased by galectins, secreted lectins capable of recognizing and clustering glycoconjugates, affecting their motility and activity. Cell surface N-glycosylation is intensively remodeled by cancer cells; however, the contribution of this phenomenon to the efficiency of treatment with cytotoxic conjugates is largely unknown. Here, we evaluated the significance of N-glycosylation for the internalization and toxicity of conjugates targeting two model receptor tyrosine kinases strongly implicated in cancer: HER2 and FGFR1. We employed three conjugates of distinct molecular architecture and specificity: Affibody<sub>HER2</sub>-vcMMAE (targeting HER2), vcMMAE-KCK-FGF1.E and T-Fc-vcMMAE (recognizing different epitopes within FGFR1). We demonstrated that inhibition of N-glycosylation reduced the cellular uptake of all conjugates tested and provided evidence for a role of the galectin network in conjugate internalization. In vitro binding studies revealed that the reduced uptake of conjugates is not due to impaired HER2 and FGFR1 binding. Importantly, we demonstrated that alteration of N-glycosylation can affect the cytotoxic potential of conjugates. Our data implicate a key role for cell surface N-glycosylation in the delivery of cytotoxic conjugates into cancer cells. |
| first_indexed | 2024-03-09T10:08:33Z |
| format | Article |
| id | doaj.art-5f5f6b7264f942bfb061fe511ef5aae2 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T10:08:33Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-5f5f6b7264f942bfb061fe511ef5aae22023-12-01T22:57:51ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-07-012315851410.3390/ijms23158514The Significance of Cell Surface N-Glycosylation for Internalization and Potency of Cytotoxic Conjugates Targeting Receptor Tyrosine KinasesMarta Poźniak0Dominika Żukowska1Aleksandra Gędaj2Mateusz Adam Krzyścik3Natalia Porębska4Małgorzata Zakrzewska5Jacek Otlewski6Łukasz Opaliński7Faculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, PolandFaculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, PolandFaculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, PolandFaculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, PolandFaculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, PolandFaculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, PolandFaculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, PolandFaculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, PolandPrecise anticancer therapies employing cytotoxic conjugates constitute a side-effect-limited, highly attractive alternative to commonly used cancer treatment modalities, such as conventional chemotherapy, radiotherapy or surgical interventions. Receptor tyrosine kinases are a large family of N-glycoproteins intensively studied as molecular targets for cytotoxic conjugates in various cancers. At the cell surface, these receptors are embedded in a dense carbohydrate layer formed by numerous plasma membrane glycoproteins. The complexity of the cell surface architecture is further increased by galectins, secreted lectins capable of recognizing and clustering glycoconjugates, affecting their motility and activity. Cell surface N-glycosylation is intensively remodeled by cancer cells; however, the contribution of this phenomenon to the efficiency of treatment with cytotoxic conjugates is largely unknown. Here, we evaluated the significance of N-glycosylation for the internalization and toxicity of conjugates targeting two model receptor tyrosine kinases strongly implicated in cancer: HER2 and FGFR1. We employed three conjugates of distinct molecular architecture and specificity: Affibody<sub>HER2</sub>-vcMMAE (targeting HER2), vcMMAE-KCK-FGF1.E and T-Fc-vcMMAE (recognizing different epitopes within FGFR1). We demonstrated that inhibition of N-glycosylation reduced the cellular uptake of all conjugates tested and provided evidence for a role of the galectin network in conjugate internalization. In vitro binding studies revealed that the reduced uptake of conjugates is not due to impaired HER2 and FGFR1 binding. Importantly, we demonstrated that alteration of N-glycosylation can affect the cytotoxic potential of conjugates. Our data implicate a key role for cell surface N-glycosylation in the delivery of cytotoxic conjugates into cancer cells.https://www.mdpi.com/1422-0067/23/15/8514N-glycosylationendocytosisgalectinscytotoxic conjugatescancer therapyRTK |
| spellingShingle | Marta Poźniak Dominika Żukowska Aleksandra Gędaj Mateusz Adam Krzyścik Natalia Porębska Małgorzata Zakrzewska Jacek Otlewski Łukasz Opaliński The Significance of Cell Surface N-Glycosylation for Internalization and Potency of Cytotoxic Conjugates Targeting Receptor Tyrosine Kinases International Journal of Molecular Sciences N-glycosylation endocytosis galectins cytotoxic conjugates cancer therapy RTK |
| title | The Significance of Cell Surface N-Glycosylation for Internalization and Potency of Cytotoxic Conjugates Targeting Receptor Tyrosine Kinases |
| title_full | The Significance of Cell Surface N-Glycosylation for Internalization and Potency of Cytotoxic Conjugates Targeting Receptor Tyrosine Kinases |
| title_fullStr | The Significance of Cell Surface N-Glycosylation for Internalization and Potency of Cytotoxic Conjugates Targeting Receptor Tyrosine Kinases |
| title_full_unstemmed | The Significance of Cell Surface N-Glycosylation for Internalization and Potency of Cytotoxic Conjugates Targeting Receptor Tyrosine Kinases |
| title_short | The Significance of Cell Surface N-Glycosylation for Internalization and Potency of Cytotoxic Conjugates Targeting Receptor Tyrosine Kinases |
| title_sort | significance of cell surface n glycosylation for internalization and potency of cytotoxic conjugates targeting receptor tyrosine kinases |
| topic | N-glycosylation endocytosis galectins cytotoxic conjugates cancer therapy RTK |
| url | https://www.mdpi.com/1422-0067/23/15/8514 |
| work_keys_str_mv | AT martapozniak thesignificanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT dominikazukowska thesignificanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT aleksandragedaj thesignificanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT mateuszadamkrzyscik thesignificanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT nataliaporebska thesignificanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT małgorzatazakrzewska thesignificanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT jacekotlewski thesignificanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT łukaszopalinski thesignificanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT martapozniak significanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT dominikazukowska significanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT aleksandragedaj significanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT mateuszadamkrzyscik significanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT nataliaporebska significanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT małgorzatazakrzewska significanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT jacekotlewski significanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases AT łukaszopalinski significanceofcellsurfacenglycosylationforinternalizationandpotencyofcytotoxicconjugatestargetingreceptortyrosinekinases |