(111)In-labeled trastuzumab (Herceptin) modified with nuclear localization sequences (NLS): an Auger electron-emitting radiotherapeutic agent for HER2/neu-amplified breast cancer.
UNLABELLED: The cytotoxicity and tumor-targeting properties of the anti-HER2/neu monoclonal antibody trastuzumab modified with peptides (CGYGPKKKRKVGG) harboring the nuclear localization sequence ([NLS] italicized) of simian virus 40 large T-antigen and radiolabeled with (111)In were evaluated. METH...
| Główni autorzy: | , , , , |
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| Format: | Journal article |
| Język: | English |
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2007
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| _version_ | 1826292694951919616 |
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| author | Costantini, D Chan, C Cai, Z Vallis, K Reilly, R |
| author_facet | Costantini, D Chan, C Cai, Z Vallis, K Reilly, R |
| author_sort | Costantini, D |
| collection | OXFORD |
| description | UNLABELLED: The cytotoxicity and tumor-targeting properties of the anti-HER2/neu monoclonal antibody trastuzumab modified with peptides (CGYGPKKKRKVGG) harboring the nuclear localization sequence ([NLS] italicized) of simian virus 40 large T-antigen and radiolabeled with (111)In were evaluated. METHODS: Trastuzumab was derivatized with sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC) for reaction with NLS-peptides and labeled with (111)In using diethylenetriaminepentaacetic acid (DTPA). The immunoreactivity of (111)In-NLS-trastuzumab was determined by its ability to displace the binding of trastuzumab to SK-BR-3 human breast cancer (BC) cells. Cellular uptake and nuclear localization were evaluated in SK-BR-3, MDA-MB-361, and MDA-MB-231 BC cells, expressing high, intermediate, or very low levels of HER2/neu, respectively, by cell fractionation and confocal microscopy. Biodistribution and nuclear uptake were compared in athymic mice bearing MDA-MB-361 xenografts. The cytotoxicity of (111)In-trastuzumab and (111)In-NLS-trastuzumab was studied by clonogenic assays, and DNA damage was assessed by probing for phosphorylated histone H2AX (gammaH2AX) foci. RESULTS: The dissociation constant for binding of (111)In-NLS-trastuzumab to SK-BR-3 cells was reduced <3-fold compared with that of (111)In-trastuzumab, demonstrating relatively preserved receptor-binding affinity. The receptor-mediated internalization of (111)In-trastuzumab in SK-BR-3, MDA-MB-361, and MDA-MB-231 cells increased significantly from 7.2% +/- 0.9%, 1.3% +/- 0.1%, and 0.2% +/- 0.05% to 14.4% +/- 1.8%, 6.3% +/- 0.2%, and 0.9% +/- 0.2% for (111)In-NLS-trastuzumab harboring 6 NLS-peptides, respectively. NLS-trastuzumab localized in the nuclei of BC cells, whereas unmodified trastuzumab remained surface-bound. Conjugation of (111)In-trastuzumab to NLS-peptides did not affect its tissue biodistribution but promoted specific nuclear uptake in MDA-MB-361 xenografts (2.4-2.9 %ID/g [percentage injected dose per gram] for (111)In-NLS-trastuzumab and 1.1 %ID/g for (111)In-trastuzumab). (111)In-NLS-trastuzumab was 5- and 2-fold more potent at killing SK-BR-3 and MDA-MB-361 cells than (111)In-trastuzumab, respectively, whereas toxicity toward MDA-MB-231 cells was minimal. (111)In-NLS-trastuzumab was 6-fold more effective at killing SK-BR-3 cells than unlabeled trastuzumab. Formation of gammaH2AX foci occurred in a greater proportion of BC cells after incubation with (111)In-NLS-trastuzumab compared with (111)In-trastuzumab or unlabeled trastuzumab. CONCLUSION: NLS-peptides routed (111)In-trastuzumab to the nucleus of HER2/neu-positive human BC cells, rendering the radiopharmaceutical lethal to the cells through the emission of nanometer-micrometer range Auger electrons. The greater cytotoxic potency of (111)In-NLS-trastuzumab compared with unlabeled trastuzumab in vitro and its favorable tumor-targeting properties in vivo suggest that it could be an effective targeted radiotherapeutic agent for HER2/neu-amplified BC in humans. |
| first_indexed | 2024-03-07T03:18:40Z |
| format | Journal article |
| id | oxford-uuid:b6b1f644-f5ed-4095-a5ad-6b6966c6a11f |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T03:18:40Z |
| publishDate | 2007 |
| record_format | dspace |
| spelling | oxford-uuid:b6b1f644-f5ed-4095-a5ad-6b6966c6a11f2022-03-27T04:42:46Z(111)In-labeled trastuzumab (Herceptin) modified with nuclear localization sequences (NLS): an Auger electron-emitting radiotherapeutic agent for HER2/neu-amplified breast cancer.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:b6b1f644-f5ed-4095-a5ad-6b6966c6a11fEnglishSymplectic Elements at Oxford2007Costantini, DChan, CCai, ZVallis, KReilly, RUNLABELLED: The cytotoxicity and tumor-targeting properties of the anti-HER2/neu monoclonal antibody trastuzumab modified with peptides (CGYGPKKKRKVGG) harboring the nuclear localization sequence ([NLS] italicized) of simian virus 40 large T-antigen and radiolabeled with (111)In were evaluated. METHODS: Trastuzumab was derivatized with sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC) for reaction with NLS-peptides and labeled with (111)In using diethylenetriaminepentaacetic acid (DTPA). The immunoreactivity of (111)In-NLS-trastuzumab was determined by its ability to displace the binding of trastuzumab to SK-BR-3 human breast cancer (BC) cells. Cellular uptake and nuclear localization were evaluated in SK-BR-3, MDA-MB-361, and MDA-MB-231 BC cells, expressing high, intermediate, or very low levels of HER2/neu, respectively, by cell fractionation and confocal microscopy. Biodistribution and nuclear uptake were compared in athymic mice bearing MDA-MB-361 xenografts. The cytotoxicity of (111)In-trastuzumab and (111)In-NLS-trastuzumab was studied by clonogenic assays, and DNA damage was assessed by probing for phosphorylated histone H2AX (gammaH2AX) foci. RESULTS: The dissociation constant for binding of (111)In-NLS-trastuzumab to SK-BR-3 cells was reduced <3-fold compared with that of (111)In-trastuzumab, demonstrating relatively preserved receptor-binding affinity. The receptor-mediated internalization of (111)In-trastuzumab in SK-BR-3, MDA-MB-361, and MDA-MB-231 cells increased significantly from 7.2% +/- 0.9%, 1.3% +/- 0.1%, and 0.2% +/- 0.05% to 14.4% +/- 1.8%, 6.3% +/- 0.2%, and 0.9% +/- 0.2% for (111)In-NLS-trastuzumab harboring 6 NLS-peptides, respectively. NLS-trastuzumab localized in the nuclei of BC cells, whereas unmodified trastuzumab remained surface-bound. Conjugation of (111)In-trastuzumab to NLS-peptides did not affect its tissue biodistribution but promoted specific nuclear uptake in MDA-MB-361 xenografts (2.4-2.9 %ID/g [percentage injected dose per gram] for (111)In-NLS-trastuzumab and 1.1 %ID/g for (111)In-trastuzumab). (111)In-NLS-trastuzumab was 5- and 2-fold more potent at killing SK-BR-3 and MDA-MB-361 cells than (111)In-trastuzumab, respectively, whereas toxicity toward MDA-MB-231 cells was minimal. (111)In-NLS-trastuzumab was 6-fold more effective at killing SK-BR-3 cells than unlabeled trastuzumab. Formation of gammaH2AX foci occurred in a greater proportion of BC cells after incubation with (111)In-NLS-trastuzumab compared with (111)In-trastuzumab or unlabeled trastuzumab. CONCLUSION: NLS-peptides routed (111)In-trastuzumab to the nucleus of HER2/neu-positive human BC cells, rendering the radiopharmaceutical lethal to the cells through the emission of nanometer-micrometer range Auger electrons. The greater cytotoxic potency of (111)In-NLS-trastuzumab compared with unlabeled trastuzumab in vitro and its favorable tumor-targeting properties in vivo suggest that it could be an effective targeted radiotherapeutic agent for HER2/neu-amplified BC in humans. |
| spellingShingle | Costantini, D Chan, C Cai, Z Vallis, K Reilly, R (111)In-labeled trastuzumab (Herceptin) modified with nuclear localization sequences (NLS): an Auger electron-emitting radiotherapeutic agent for HER2/neu-amplified breast cancer. |
| title | (111)In-labeled trastuzumab (Herceptin) modified with nuclear localization sequences (NLS): an Auger electron-emitting radiotherapeutic agent for HER2/neu-amplified breast cancer. |
| title_full | (111)In-labeled trastuzumab (Herceptin) modified with nuclear localization sequences (NLS): an Auger electron-emitting radiotherapeutic agent for HER2/neu-amplified breast cancer. |
| title_fullStr | (111)In-labeled trastuzumab (Herceptin) modified with nuclear localization sequences (NLS): an Auger electron-emitting radiotherapeutic agent for HER2/neu-amplified breast cancer. |
| title_full_unstemmed | (111)In-labeled trastuzumab (Herceptin) modified with nuclear localization sequences (NLS): an Auger electron-emitting radiotherapeutic agent for HER2/neu-amplified breast cancer. |
| title_short | (111)In-labeled trastuzumab (Herceptin) modified with nuclear localization sequences (NLS): an Auger electron-emitting radiotherapeutic agent for HER2/neu-amplified breast cancer. |
| title_sort | 111 in labeled trastuzumab herceptin modified with nuclear localization sequences nls an auger electron emitting radiotherapeutic agent for her2 neu amplified breast cancer |
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