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ARHGAP–RhoA signaling provokes homotypic adhesion-triggered cell death of metastasized diffuse-type gastric cancer

Abstract

Genetic alteration of Rho GTPase-activating proteins (ARHGAP) and GTPase RhoA is a hallmark of diffuse-type gastric cancer and elucidating its biological significance is critical to comprehensively understanding this malignancy. Here, we report that gene fusions of ARHGAP6/ARHGAP26 are frequent genetic events in peritoneally-metastasized gastric and pancreatic cancer. From the malignant ascites of patients, we established gastric cancer cell lines that spontaneously gain hotspot RHOA mutations or four different ARHGAP6/ARHGAP26 fusions. These alterations critically downregulate RhoA–ROCK–MLC2 signaling, which elicits cell death. Omics and functional analyses revealed that the downstream signaling initiates actin stress fibers and reinforces intercellular junctions via several types of catenin. E-cadherin-centered homotypic adhesion followed by lysosomal membrane permeabilization is a pivotal mechanism in cell death. These findings support the tumor-suppressive nature of ARHGAP–RhoA signaling and might indicate a new avenue of drug discovery against this refractory cancer.

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Fig. 1: Identification of ARHGAP6/26 fusions and mutated RHOA in established DGC cell lines.
Fig. 2: RhoA Y42C downregulates ROCK-MLC2 signaling and escapes from cell death.
Fig. 3: RhoA elicits lysosomal leakage via E-cadherin-mediated homotypic adhesion.
Fig. 4: ARHGAP26 fusions inhibit RhoA and evade cell death.
Fig. 5: Loss of ARHGAP26 fusions activated RhoA-ROCK signaling and elicit E-cadherin-mediated homotypic adhesion.
Fig. 6: RP2-ARHGAP6 fusion give DGC cells invasiveness and caner stemness.
Fig. 7: Whole picture of dysregulated ARHGAP-RhoA signaling in DGC cells.

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Acknowledgements

This research was supported by the National Institute of Biomedical Innovation (Program ID10-41), by the National Cancer Center Research and Development Fund (28-A-11, 29-A-2, and 2020-J-2), by AMED (Japan Agency for Medical Research and Development, JP20ck0106519), and by Astellas Pharma, Inc (CH27066). We thank H. Nikki March, PhD, from Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript.

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MK planned the study, performed experiments, analyzed the data, and wrote the manuscript; FC, RK, and MA performed the experiments; HI, HSak, and KT carried out next-generation sequencing and analyzed the data; TF, HM, NB, and KM collected clinical samples; TY, FT, and HSas conceived and planned the study, analyzed the data, and wrote the manuscript. All authors have read and approved the final version of the manuscript.

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Correspondence to Masayuki Komatsu or Hiroki Sasaki.

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Komatsu, M., Ichikawa, H., Chiwaki, F. et al. ARHGAP–RhoA signaling provokes homotypic adhesion-triggered cell death of metastasized diffuse-type gastric cancer. Oncogene 41, 4779–4794 (2022). https://doi.org/10.1038/s41388-022-02469-6

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