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Oncogene addiction to GNAS in GNASR201 mutant tumors

Oncogene volume 41, pages 4159–4168 (2022)Cite this article

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Abstract

The GNASR201 gain-of-function mutation is the single most frequent cancer-causing mutation across all heterotrimeric G proteins, driving oncogenesis in various low-grade/benign gastrointestinal and pancreatic tumors. In this study, we investigated the role of GNAS and its product Gαs in tumor progression using peritoneal models of colorectal cancer (CRC). GNAS was knocked out in multiple CRC cell lines harboring GNASR201C/H mutations (KM12, SNU175, SKCO1), leading to decreased cell-growth in 2D and 3D organoid models. Nude mice were peritoneally injected with GNAS-knockout KM12 cells, leading to a decrease in tumor growth and drastically improved survival at 7 weeks. Supporting these findings, GNAS overexpression in LS174T cells led to increased cell-growth in 2D and 3D organoid models, and increased tumor growth in PDX mouse models. GNAS knockout decreased levels of cyclic AMP in KM12 cells, and molecular profiling identified phosphorylation of β-catenin and activation of its targets as critical downstream effects of mutant GNAS signaling. Supporting these findings, chemical inhibition of both PKA and β-catenin reduced growth of GNAS mutant organoids. Our findings demonstrate oncogene addiction to GNAS in peritoneal models of GNASR201C/H tumors, which signal through the cAMP/PKA and Wnt/β-catenin pathways. Thus, GNAS and its downstream mediators are promising therapeutic targets for GNAS mutant tumors.

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Fig. 1: Incidence of GNAS mutations in pan-cancer.
Fig. 2: Evaluation of GNAS knockout and overexpressing CRC cell lines.
Fig. 3: GNAS promotes tumor formation from KM12 and LS174T cells in the peritoneum of NSG mice.
Fig. 4: GNAS signals through the cAMP-PKA axis.
Fig. 5: β-catenin is a downstream mediator of mutant GNAS signaling.

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Data availability

All data supporting the findings of this study are available within the paper and its supplementary information files, gene expression data will be uploaded to GEO. Data generated for this study are available through the Gene Expression Omnibus (GEO). Accession number: GSE208278.

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Acknowledgements

This work was supported by the National Cancer Institute (L30 CA171000 and K22 CA234406 to JPS, and the Cancer Center Support Grant (P30 CA016672), the Cancer Prevention & Research Institute of Texas (RR180035 to JPS, JPS is a CPRIT Scholar in Cancer Research), and the Col. Daniel Connelly Memorial Fund. We thank the following core facilities at MD Anderson Cancer Center for their services used in this study: Reverse Phase Protein Array Core (Supported by NCI grant # CA16672 and #R50CA221675), Advanced Technology Genomics Core (Supported by NCI Grant CA016672(ATGC)), Biospecimen Extraction Facility for sample processing and DNA/RNA/protein extraction, and Small Animal Imaging Facility (Supported by the Cancer Center Support Grant CA16672) for in vivo live imaging. In addition, data generated by TCGA Research Network: https://www.cancer.gov/tcga was used in this publication. We also acknowledge Dr Kenna Shaw and data integration and clinical team members from the Sheikh Khalifa Bin Zayed Al Nahyan Institute for Personalized Cancer Therapy, supported by the Khalifa Bin Zayed Al Nahyan Foundation, for building and maintaining the MOCLIP database.

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  1. These authors contributed equally: Aditya More, Ichiaki Ito, Valsala Haridas

Authors and Affiliations

  1. Department of GI Medical Oncology, MD Anderson Cancer Center, Houston, TX, 77030, USA

    Aditya More, Ichiaki Ito, Valsala Haridas, Saikat Chowdhury, Yue Gu, Princess Dickson & John Paul Shen

  2. Department of Veterinary Medicine & Surgery, MD Anderson Cancer Center, Houston, TX, 77030, USA

    Natalie Fowlkes

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Contributions

AM, II, and VH conceived, designed, and performed all experiments and wrote the paper. SC and YG performed computational analysis and helped write the paper. PD performed animal experiments, and NF quantified and analyzed immunohistochemistry data. JPS conceived, designed and supervised the study, and wrote the paper.

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Correspondence to John Paul Shen.

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More, A., Ito, I., Haridas, V. et al. Oncogene addiction to GNAS in GNASR201 mutant tumors. Oncogene 41, 4159–4168 (2022). https://doi.org/10.1038/s41388-022-02388-6

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