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SCAMP3 is a mutant EGFR phosphorylation target and a tumor suppressor in lung adenocarcinoma

Oncogene volume 40, pages 3331–3346 (2021)Cite this article

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Abstract

Mutations in the epidermal growth factor receptor (EGFR) tyrosine kinase domain constitutively activate EGFR resulting in lung tumorigenesis. Activated EGFR modulates downstream signaling by altering phosphorylation-driven interactions that promote growth and survival. Secretory carrier membrane proteins (SCAMPs) are a family of transmembrane proteins that regulate recycling of receptor proteins, including EGFR. The potential role of SCAMPs in mutant EGFR function and tumorigenesis has not been elucidated. Using quantitative mass-spectrometry-based phosphoproteomics, we identified SCAMP3 as a target of mutant EGFRs in lung adenocarcinoma and sought to further investigate the role of SCAMP3 in the regulation of lung tumorigenesis. Here we show that activated EGFR, either directly or indirectly phosphorylates SCAMP3 at Y86 and this phosphorylation increases the interaction of SCAMP3 with both wild-type and mutant EGFRs. SCAMP3 knockdown increases lung adenocarcinoma cell survival and increases xenograft tumor growth in vivo, demonstrating a tumor suppressor role of SCAMP3 in lung tumorigenesis. The tumor suppressor function is a result of SCAMP3 promoting EGFR degradation and attenuating MAP kinase signaling pathways. SCAMP3 knockdown also increases multinucleated cells in culture, suggesting that SCAMP3 is required for efficient cytokinesis. The enhanced growth, increased colony formation, reduced EGFR degradation and multinucleation phenotype of SCAMP3-depleted cells were reversed by re-expression of wild-type SCAMP3, but not SCAMP3 Y86F, suggesting that Y86 phosphorylation is critical for SCAMP3 function. Taken together, the results of this study demonstrate that SCAMP3 functions as a novel tumor suppressor in lung cancer by modulating EGFR signaling and cytokinesis that is partly Y86 phosphorylation-dependent.

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Fig. 1: Identification of EGFR-regulated SCAMP3 phosphotyrosine sites by quantitative mass spectrometry.
Fig. 2: SCAMP3 phosphorylation is EGFR-dependent and enhances SCAMP3–EGFR interaction.
Fig. 3: SCAMP3 depletion accelerates tumor growth in human lung adenocarcinoma cells expressing EGFRWT.
Fig. 4: SCAMP3 depletion accelerates tumor growth in human lung adenocarcinoma cells expressing EGFRL858R/T790M and may decrease sensitivity to EGFR-TKI.
Fig. 5: SCAMP3 enhances EGFR degradation upon EGF stimulation.
Fig. 6: SCAMP3 depletion alters nuclear number and morphology in lung adenocarcinoma.

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Acknowledgements

We thank Gaga Geneti for the assistance with the mice handling and treatments. This research was supported by the NIH Intramural Research Program, Center of Cancer Research, National Cancer Institute.

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  1. Udayan Guha

    Present address: Bristol Myers Squibb, Lawrenceville, NJ, USA

Authors and Affiliations

  1. Thoracic and GI Malignancies Branch, Center for Cancer Research, NCI, NIH, Bethesda, MD, USA

    Abhilash Venugopalan, Matthew Lynberg, Constance M. Cultraro, Khoa Dang P. Nguyen, Xu Zhang, Maryam Waris, Noelle Dayal, Asebot Abebe, Tapan K. Maity & Udayan Guha

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Correspondence to Abhilash Venugopalan or Udayan Guha.

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UG has a clinical trial agreement (CTA) with AstraZeneca and received research funding from AstraZeneca, Esanex and Aurigene. The other authors declare no competing interests.

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Venugopalan, A., Lynberg, M., Cultraro, C.M. et al. SCAMP3 is a mutant EGFR phosphorylation target and a tumor suppressor in lung adenocarcinoma. Oncogene 40, 3331–3346 (2021). https://doi.org/10.1038/s41388-021-01764-y

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