Targeted Delivery of Deoxycytidine Kinase to Her2-Positive Cells Enhances the Efficacy of the Nucleoside Analog Fludarabine
| Autor: | Ying Su, Sujatha P. Koduvayur, Arnon Lavie, Brian K. Kay |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Receptor ErbB-2 Cancer Treatment Glycobiology lcsh:Medicine Apoptosis Biochemistry Cell Fusion Spectrum Analysis Techniques Drug Delivery Systems Medicine and Health Sciences Tumor Cells Cultured Phosphorylation lcsh:Science Ovarian Neoplasms Multidisciplinary Pro-Drugs Conjugated Proteins Drugs Nucleosides Deoxycytidine kinase Flow Cytometry Combined Modality Therapy Glycosylamines 3. Good health Fludarabine Cell killing Oncology DARPin Cell Processes Spectrophotometry Female Cytophotometry Vidarabine Research Article medicine.drug Cell Binding Cell Physiology Breast Neoplasms Biology Research and Analysis Methods 03 medical and health sciences Deoxycytidine Kinase medicine Humans Cell Proliferation Pharmacology Nucleoside analogue Cell growth lcsh:R Biology and Life Sciences Proteins Cell Biology 030104 developmental biology Cancer cell Cancer research lcsh:Q Nucleoside |
| Zdroj: | PLoS ONE PLoS ONE, Vol 11, Iss 6, p e0157114 (2016) |
| ISSN: | 1932-6203 |
| Popis: | Cytotoxic drugs, such as nucleoside analogs and toxins, commonly suffer from off-target effects. One approach to mitigate this problem is to deliver the cytotoxic drug selectively to the intended site. While for toxins this can be achieved by conjugating the cell-killing moiety to a targeting moiety, it is not an option for nucleoside analogs, which rely on intracellular enzymes to convert them to their active triphosphorylated form. To overcome this limitation, and achieve site-targeted activation of nucleoside analogs, we fused the coding region of a prodrug-activating enzyme, deoxycytidine kinase (dCK), to affinity reagents that bind to the Her2 cell surface protein. We evaluated dCK fusions to an anti-Her2 affibody and Designed Ankyrin Repeat Protein (DARPin) for their ability to kill cancer cells by promoting the activation of the nucleoside analog fludarabine. Cell staining and flow cytometry experiments with three Her2 positive cancer cell lines (BT-474-JB, JIMT-1 and SK-OV-3) indicate dCK fusions binding and cellular internalization. In contrast, these reagents bind only weakly to the Her2 negative cell line, MCF-7. Cell proliferation assays indicate that SK-OV-3 and BT-474-JB cell lines exhibit significantly reduced proliferation rates when treated with targeting-module fused dCK and fludarabine, compared to fludarabine alone. These findings demonstrate that we have succeeded in delivering active dCK into the Her2-positive cells, thereby increasing the activation of fludarabine, which ultimately reduces the dose of nucleoside analog needed for cell killing. This strategy may help establish the therapeutic index required to differentiate between healthy tissues and cancer cells. |
| Databáze: | OpenAIRE |
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