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The G Protein–Coupled Receptor PAC1 Regulates Transactivation of the Receptor Tyrosine Kinase HER3

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Abstract

Peptide G protein–coupled receptors (GPCRs) for pituitary adenylate cyclase activating polypeptide (PACAP) regulate the growth of non-small cell lung cancer (NSCLC) cells. PACAP binds with high affinity to PAC1, which causes transactivation of receptor tyrosine kinases (RTK) for the EGFR and HER2 but its effect on HER3 is unknown. Using 3 NSCLC cell lines (NCI-H358, NCI-H441, and Calu-3), proteins for EGFR, HER2, HER3, and PAC1 were detected. The increase in EGFR tyrosine phosphorylation caused by PACAP was blocked by the EGFR tyrosine kinase inhibitor (TKI) gefitinib, or PACAP(6-38), a PAC1 antagonist. The increase in HER2 tyrosine phosphorylation caused by PACAP was inhibited by trastuzumab, a monoclonal antibody (mAb) for HER2, or PACAP(6-38). The increase in HER3 tyrosine phosphorylation caused by PACAP was inhibited by HER3 mAb3481 or PACAP(6-38). Immunoprecipitation experiments indicated the PACAP addition to Calu-3 cells resulted in the formation of EGFR/HER3 and HER2/HER3 heterodimers. Addition of the HER3 agonist neuregulin (NRG)-1 increased HER3 tyrosine phosphorylation in non-small-cell lung cancer (NSCLC) cells. PACAP or NRG-1 increased the proliferation of NSCLC cells, whereas PACAP(6-38), gefitinib, trastuzumab, or mAb3481 inhibited proliferation. The results indicate that PAC1 regulates the proliferation of NSCLC cells as a result of transactivation of the EGFR, HER2, and HER3.

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Acknowledgments

The authors thank Dr. Irene Ramos-Gonzalez for the helpful discussions. This research is supported by the intramural programs of the NCI and NIDDK of NIH. This research was presented at the 14th Annual meeting on PACAP/VIP and related peptides in 2019 at Los Angeles, CA.

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Authors and Affiliations

  1. Department of Health and Human Services, National Institutes of Health, National Cancer Institute, Center for Cancer Training, 9609 Medical Center Drive, Room 2W-340, Bethesda, MD, 20892, USA

    Terry W. Moody

  2. National Institute of Diabetes, Digestive and Kidney Disease, Digestive Diseases Branch, Bethesda, MD, 20892, U.S.A.

    Lingaku Lee & Robert T. Jensen

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  1. Terry W. Moody
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  2. Lingaku Lee
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  3. Robert T. Jensen
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The research was conducted by Terry Moody and Lingaku Lee. The manuscript was written by Terry Moody and Robert Jensen.

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Correspondence to Terry W. Moody.

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Moody, T.W., Lee, L. & Jensen, R.T. The G Protein–Coupled Receptor PAC1 Regulates Transactivation of the Receptor Tyrosine Kinase HER3. J Mol Neurosci 71, 1589–1597 (2021). https://doi.org/10.1007/s12031-020-01711-8

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