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Development of a MET-targeted single-chain antibody fragment as an anti-oncogene targeted therapy for breast cancer

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Abstract

The usage of monoclonal antibodies (mAbs) and antibody fragments, as a matter associated with the biopharmaceutical industry, is increasingly growing. Harmonious with this concept, we designed an exclusive modeled single-chain variable fragment (scFv) against mesenchymal-epithelial transition (MET) oncoprotein. This scFv was newly developed from Onartuzumab sequence by gene cloning, and expression using bacterial host. Herein, we examined its preclinical efficacy for the reduction of tumor growth, invasiveness and angiogenesis in vitro and in vivo. Expressed anti-MET scFv demonstrated high binding capacity (48.8%) toward MET-overexpressing cancer cells. The IC50 value of anti-MET scFv against MET-positive human breast cancer cell line (MDA-MB-435) was 8.4 µg/ml whereas this value was measured as 47.8 µg/ml in MET-negative cell line BT-483. Similar concentrations could also effectively induce apoptosis in MDA-MB-435 cancer cells. Moreover, this antibody fragment could reduce migration and invasion in MDA-MB-435 cells. Grafted breast tumors in Balb/c mice showed significant tumor growth suppression as well as reduction of blood-supply in response to recombinant anti-MET treatment. Histopathology and immunohistochemical assessments revealed higher rate of response to therapy. In our study, we designed and synthetized a novel anti-MET scFv which could effectively suppress MET-overexpressing breast cancer tumors.

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Data Availability

The authors declare that all data supporting the findings of current study are provided within the manuscript.

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Acknowledgements

We would thank Dr. Vahid Siavashi and Mr. Alireza Ghovati, Department of Clinical Pathology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran. for their technical assistance.

Funding

This work was supported by Research Council, University of Tehran. Specialized research grant for Doctoral Program of Higher Education, Faculty of Veterinary Medicine (7508017/6/23).

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Authors and Affiliations

  1. Department of Clinical Pathology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

    Rana Vafaei, Zohreh Khaki & Seyed Mahdi Nassiri

  2. Recombinant Proteins Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR, Tehran, Iran

    Rana Vafaei, Malihe Salehi, Neda Jalili, Shima Moradi Kalbolandi, Roya Mirzaei & Leila Farahmand

  3. Department of Surgery and Radiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

    Mohammad Reza Esmailinejad & Alireza Vajhi

  4. Cancer Biology Research Center, Cancer Institute of Iran, Tehran University of Medical Sciences, Tehran, Iran

    Ahad Muhammadnajad

  5. Shahid Bahonar University of Kerman, Department of Clinical Sciences, Faculty of Veterinary Medicine, Kerman, Iran

    Mohammad Reza Esmailinejad

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  1. Rana Vafaei
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  2. Zohreh Khaki
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Contributions

L. F. developed the methodology; Z. Kh. and R.V. conducted the experiments, contributed to the data analysis and wrote the manuscript; M. S., N. J and S. M assisted the experiments; M. R. EN. and A.V. conducted ultrasonography; S. M. N. contributed to scientific assist and revised the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Zohreh Khaki or Leila Farahmand.

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Ethics approval and consent to participate

This project was in accordance with the national norms, the ethical principles and standards for conducting medical research in Iran and evaluated by Motamed Cancer Institute-Academic Centre for Education, Culture and Research. This institution performed its reviews based on United States Public Health Service (USPHS) regulations and applicable federal and local laws. Protocols were also approved by the Animal Care Committee of the University of Tehran (7508017623).

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Vafaei, R., Khaki, Z., Salehi, M. et al. Development of a MET-targeted single-chain antibody fragment as an anti-oncogene targeted therapy for breast cancer. Invest New Drugs 41, 226–239 (2023). https://doi.org/10.1007/s10637-023-01354-7

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