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Differential tumor inhibitory effects induced by HER3 extracellular subdomain-specific mouse monoclonal antibodies

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Abstract

Purpose

The therapeutic potential of targeting the human epidermal growth factor receptor-3 (ErbB3/HER3) has long been ignored due to impaired tyrosine kinase function and low expression level in tumor cells compared with EGFR and HER2. Although recent investigations have explored the potential benefit of HER3 targeting and several anti-HER3 agents have been developed, there is still a critical need to design and produce more efficient therapeutics. This study was designed to develop tumor inhibitory monoclonal antibodies (MAbs) against different extracellular subdomains of HER3.

Methods

Distinct extracellular subdomains of HER3 (DI+II and DIII+IV) were utilized to produce MAbs by hybridoma technology. Biochemical and functional characteristics of these MAbs were then investigated by various methodologies, including immunoblotting, flow cytometry, cell proliferation, cell signaling, and enzyme-linked immunosorbent assays.

Results

Four anti-DI+II and six anti-DIII+IV MAbs were obtained, selected based on their ability to bind recombinant full HER3 extracellular domain (ECD). Our data showed that only one anti-DI+II and four anti-DIII+IV MAbs recognized the native form of HER3 by immunoblotting. Four MAbs recognized the membranous HER3 by flow cytometry leading to induction of different levels of receptor internalization and subsequent degradation. Results of cell proliferation assays using these MAbs indicated that they differentially inhibited proliferation of HER3-expressing cancer cells and showed considerable synergistic effects in combination with trastuzumab. Selected MAb with the highest inhibitory effect significantly inhibited the phosphorylation of AKT and ERK1/2 molecules.

Conclusion

Some of the anti-HER3 MAbs produced in this study displayed tumor inhibitory function and may be considered promising candidates for future HER3-targeted cancer therapy.

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Availability of data and materials

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

Code availability

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Acknowledgements

The authors wish to thank Mohammad Ali Judaki and Akram Sadat Majidian for technical support. This study was partially supported by the National Institute for Medical Research Development of Iran (NIMAD) under Grant number 971145 and Tehran University of Medical Sciences (TUMS) under Grant number 41447.

Funding

This study was partially supported by the National Institute for Medical Research Development of Iran (NIMAD) under Grant number 971145 and Tehran University of Medical Sciences (TUMS) under Grant number 41447.

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Authors and Affiliations

  1. Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

    Danesh Hassani, Samaneh Mansouri-Fard, Maryam Mobini, Forough Golsaz-Shirazi, Mohammad Mehdi Amiri & Fazel Shokri

  2. Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran

    Mahmood Jeddi-Tehrani, Parisa Yousefi & Hengameh Ahmadi-Zare

Authors
  1. Danesh Hassani
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  2. Mahmood Jeddi-Tehrani
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  3. Parisa Yousefi
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  4. Samaneh Mansouri-Fard
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  5. Maryam Mobini
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  6. Hengameh Ahmadi-Zare
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  7. Forough Golsaz-Shirazi
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  8. Mohammad Mehdi Amiri
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  9. Fazel Shokri
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Contributions

DH: contributed to the experimental design, performance of the experiments, acquisition, analysis, and interpretation of data, and writing the manuscript. MJT: contributed to the study conceptualization and reviewing and revising the manuscript. PY: contributed to the performance of cell fusion experiments. SMF: contributed to the performance of the purification experiments. MM: contributed to the performance of the flow cytometry experiments and analysis of data. HAZ: contributed to the performance of the serological experiments. FGS: contributed to the experimental design and reviewing and revising the manuscript. MMA: contributed to the study conceptualization, project administration, experimental design, and reviewing and revising the manuscript. FS: contributed to the study conceptualization, project administration, experimental design, analysis and interpretation of data, and reviewing and revising the manuscript.

Corresponding authors

Correspondence to Mohammad Mehdi Amiri or Fazel Shokri.

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Mice were housed and handled in accordance with all applicable international and/or institutional guidelines concerning the care and use of laboratory animals. All experiments involving animal use were approved by the research ethics committee of Tehran University of Medical Sciences (Code: IR.TUMS.SPH.REC.1397.320).

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Hassani, D., Jeddi-Tehrani, M., Yousefi, P. et al. Differential tumor inhibitory effects induced by HER3 extracellular subdomain-specific mouse monoclonal antibodies. Cancer Chemother Pharmacol 89, 347–361 (2022). https://doi.org/10.1007/s00280-021-04390-3

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