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Blockade of CHRNB2 signaling with a therapeutic monoclonal antibody attenuates the aggressiveness of gastric cancer cells

Oncogene volume 40, pages 5495–5504 (2021)Cite this article

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Abstract

Here, we evaluated the therapeutic potential of antibodies (Abs) targeting cholinergic receptor nicotinic beta 2 subunit (CHRNB2) in gastric cancer. To investigate the effects of these Abs on malignant phenotypes in vitro and in mouse xenograft models, we generated gene knockouts through genome editing, performed RNA interference-mediated knockdown of gene expression, and ectopically expressed CHRNB2 in gastric cancer cells. The effects of anti-CHRNB2 Abs on the proliferation of cancer cells were evaluated both in vitro and in vivo. We determined the effects of Chrnb2 deficiency on mice and the clinical significance of CHRNB2 expression in gastric cancer clinical specimens. Knockdown of CHRNB2 attenuated gastric cancer cell proliferation, whereas forced overexpression of CHRNB2 increased cell proliferation. Knockout of CHRNB2 significantly influenced cell survival and functions associated with metastasis. The effects of polyclonal Abs targeting the C- and N-termini of CHRNB2 guided the development of anti-CHRNB2 monoclonal Abs that inhibited the growth of gastric cancer cells in vitro and in vivo. Pathway analysis revealed that CHRNB2 interfered with signaling through the PI3K-AKT and JAK-STAT pathways. Chrnb2-deficient mice exhibited normal reproduction, organ functions, and motor functions. CHRNB2 regulates multiple oncological phenotypes associated with metastasis, and blockade of CHRNB2 expression using specific Abs shows promise for controlling metastasis in gastric cancer.

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Fig. 1: Expression of CHRNB2 in gastric cancer cell lines and the effect of CHRNB2 knockdown, knockout, or overexpression on cell phenotypes.
Fig. 2: Effects of CHRNB2 knockout on the survival of gastric cancer cell lines.
Fig. 3: Effects of CHRNB2 knockout on the malignant phenotypes and drug resistance of gastric cancer cell lines.
Fig. 4: Generation and effects of anti-CHRNB2 pAbs and mAbs.
Fig. 5: Intracellular signal transduction pathways associated with CHRNB2.
Fig. 6: Characterization of Chrnb2-deficient mice.
Fig. 7: CHRNB2 expression in patient gastric tissues.

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Acknowledgements

We thank Edanz Group (https://en-author-services.edanz.com/ac) and the SPRINGER NATURE English Language Editing service (https://authorservices.springernature.com/language-editing/) for editing a draft of this manuscript. This work was supported by a Grant-in-Aid for Scientific Research (B, 20H03750), the Japan Agency for Medical Research and Development (17lm0203005j0001, 2017); the Center for Advanced Medicine and Clinical Research at Nagoya University (2018); a grant (2018) from the Japan Research Foundation for Clinical Pharmacology, Foundation for Promotion of Cancer Research, Kobayashi Foundation for Cancer Research; and the Mochida Memorial Foundation for Medical and Pharmaceutical Research.

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Authors and Affiliations

  1. Department of Gastroenterological Surgery (Surgery II), Nagoya University Graduate School of Medicine, Nagoya, Japan

    Mitsuro Kanda, Dai Shimizu, Shunsuke Nakamura, Koichi Sawaki, Shinichi Umeda, Takashi Miwa, Haruyoshi Tanaka, Yoshikuni Inokawa, Norifumi Hattori, Masamichi Hayashi, Chie Tanaka, Goro Nakayama & Yasuhiro Kodera

  2. Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan

    Yohei Iguchi & Masahisa Katsuno

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Contributions

MK: data acquisition and analysis, interpretation of data, and drafting the manuscript. SN, KS, SU, TM, HT, and DS: acquisition and interpretation of data, manuscript revision. YI, NH, MH, CT, GN, and YI: material support, generation of data. YK and MK: study concept and design, study supervision, interpretation of data, and revision of the manuscript. MK had full access to all data and takes full responsibility for the veracity of the data and statistical analysis.

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Correspondence to Mitsuro Kanda.

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Kanda, M., Shimizu, D., Nakamura, S. et al. Blockade of CHRNB2 signaling with a therapeutic monoclonal antibody attenuates the aggressiveness of gastric cancer cells. Oncogene 40, 5495–5504 (2021). https://doi.org/10.1038/s41388-021-01945-9

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