Suppression of Breast Cancer Cell Proliferation by Selective Single-Domain Antibody for Intracellular STAT3
Background: The serendipitous discovery of heavy-chain antibodies devoid of light chains in camelids and the subsequent development of VHHs (variable region of camelid heavy chain) have provided a very important tool for research and possibly for therapeutics. In this study, we synthesized single-do...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2018-01-01
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| Series: | Breast Cancer: Basic and Clinical Research |
| Online Access: | https://doi.org/10.1177/1178223417750858 |
| _version_ | 1811335274459824128 |
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| author | Sunanda Singh Genoveva Murillo Dong Chen Ashutosh S Parihar Rajendra G Mehta |
| author_facet | Sunanda Singh Genoveva Murillo Dong Chen Ashutosh S Parihar Rajendra G Mehta |
| author_sort | Sunanda Singh |
| collection | DOAJ |
| description | Background: The serendipitous discovery of heavy-chain antibodies devoid of light chains in camelids and the subsequent development of VHHs (variable region of camelid heavy chain) have provided a very important tool for research and possibly for therapeutics. In this study, we synthesized single-domain 15-kDa antibody SBT-100 (anti-STAT3 B VHH13) against human STAT3 (signal transducer and activator of transcription) that binds selectively to STAT3 and suppresses the function of phosphorylated STAT3 (p-STAT3). Methods: Single-chain VHH nanobodies were generated by immunizing camelid with humanized STAT3. Commercially available breast cancer cell lines including MDA-MB-231, MDA-MB-468, MDA-MB-453, MCF-7, and BT474 were used. Cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The association of anti-STAT3 B VHH13 with STAT3 and p-STAT3 was determined by immunoprecipitation and Western blot analyses. The efficacy of SBT-100 on the growth of MDA-MB-231 xenografts in vivo was determined using athymic mice. Statistical significance for cell proliferation was determined using analysis of variance. If a significant difference ( P < .05) was observed, then Tukey-Kramer multiple comparison test was conducted. Results: SBT-100 suppressed cell proliferation of triple-negative breast cancer cells ( P < .01) as well as provided significant inhibition of tumor growth ( P < .05) in a xenograft model without any toxicity. Results are presented to show that anti-STAT3 B VHH13 selectively binds to STAT3 suggesting that the effects were mediated by inhibiting STAT3. Conclusions: A very large number of human malignancies and benign diseases have constitutive STAT3 activation. Therefore, the results described here suggest that anti-STAT3 B VHH13 can be developed for therapeutic intervention for cancer cells expressing STAT3 or p-STAT3. |
| first_indexed | 2024-04-13T17:21:46Z |
| format | Article |
| id | doaj.art-d2f65828cb614510aa53aa26020b6b2b |
| institution | Directory Open Access Journal |
| issn | 1178-2234 |
| language | English |
| last_indexed | 2024-04-13T17:21:46Z |
| publishDate | 2018-01-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Breast Cancer: Basic and Clinical Research |
| spelling | doaj.art-d2f65828cb614510aa53aa26020b6b2b2022-12-22T02:37:58ZengSAGE PublishingBreast Cancer: Basic and Clinical Research1178-22342018-01-011210.1177/1178223417750858Suppression of Breast Cancer Cell Proliferation by Selective Single-Domain Antibody for Intracellular STAT3Sunanda Singh0Genoveva Murillo1Dong Chen2Ashutosh S Parihar3Rajendra G Mehta4Singh Biotechnology and Tampa Bay Technology Incubator, University of South Florida, Tampa Bay, FL, USAIIT Research Institute, Chicago, IL, USACreative Biolabs, Shirley, NY, USASingh Biotechnology and Tampa Bay Technology Incubator, University of South Florida, Tampa Bay, FL, USAIIT Research Institute, Chicago, IL, USABackground: The serendipitous discovery of heavy-chain antibodies devoid of light chains in camelids and the subsequent development of VHHs (variable region of camelid heavy chain) have provided a very important tool for research and possibly for therapeutics. In this study, we synthesized single-domain 15-kDa antibody SBT-100 (anti-STAT3 B VHH13) against human STAT3 (signal transducer and activator of transcription) that binds selectively to STAT3 and suppresses the function of phosphorylated STAT3 (p-STAT3). Methods: Single-chain VHH nanobodies were generated by immunizing camelid with humanized STAT3. Commercially available breast cancer cell lines including MDA-MB-231, MDA-MB-468, MDA-MB-453, MCF-7, and BT474 were used. Cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The association of anti-STAT3 B VHH13 with STAT3 and p-STAT3 was determined by immunoprecipitation and Western blot analyses. The efficacy of SBT-100 on the growth of MDA-MB-231 xenografts in vivo was determined using athymic mice. Statistical significance for cell proliferation was determined using analysis of variance. If a significant difference ( P < .05) was observed, then Tukey-Kramer multiple comparison test was conducted. Results: SBT-100 suppressed cell proliferation of triple-negative breast cancer cells ( P < .01) as well as provided significant inhibition of tumor growth ( P < .05) in a xenograft model without any toxicity. Results are presented to show that anti-STAT3 B VHH13 selectively binds to STAT3 suggesting that the effects were mediated by inhibiting STAT3. Conclusions: A very large number of human malignancies and benign diseases have constitutive STAT3 activation. Therefore, the results described here suggest that anti-STAT3 B VHH13 can be developed for therapeutic intervention for cancer cells expressing STAT3 or p-STAT3.https://doi.org/10.1177/1178223417750858 |
| spellingShingle | Sunanda Singh Genoveva Murillo Dong Chen Ashutosh S Parihar Rajendra G Mehta Suppression of Breast Cancer Cell Proliferation by Selective Single-Domain Antibody for Intracellular STAT3 Breast Cancer: Basic and Clinical Research |
| title | Suppression of Breast Cancer Cell Proliferation by Selective Single-Domain Antibody for Intracellular STAT3 |
| title_full | Suppression of Breast Cancer Cell Proliferation by Selective Single-Domain Antibody for Intracellular STAT3 |
| title_fullStr | Suppression of Breast Cancer Cell Proliferation by Selective Single-Domain Antibody for Intracellular STAT3 |
| title_full_unstemmed | Suppression of Breast Cancer Cell Proliferation by Selective Single-Domain Antibody for Intracellular STAT3 |
| title_short | Suppression of Breast Cancer Cell Proliferation by Selective Single-Domain Antibody for Intracellular STAT3 |
| title_sort | suppression of breast cancer cell proliferation by selective single domain antibody for intracellular stat3 |
| url | https://doi.org/10.1177/1178223417750858 |
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