Abstract
Background
Mutations in GNAS drive pancreatic tumorigenesis and frequently occur in intraductal papillary mucinous neoplasm (IPMN); however, their value as a therapeutic target is yet to be determined. This study aimed at evaluating the involvement of mutant GNAS in tumor aggressiveness in established pancreatic cancer.
Methods
CRISPR/Cas9-mediated GNAS R201H silencing was performed using human primary IPMN-associated pancreatic cancer cells. The role of oncogenic GNAS in tumor maintenance was evaluated by conducting cell culture and xenograft experiments, and western blotting and transcriptome analyses were performed to uncover GNAS-driven signatures.
Results
Xenografts of GNAS wild-type cells were characterized by a higher Ki-67 labeling index relative to GNAS-mutant cells. Phenotypic alterations in the GNAS wild-type tumors resulted in a significant reduction in mucin production accompanied by solid with massive stromal components. Transcriptional profiling suggested an apparent conflict of mutant GNAS with KRAS signaling. A significantly higher Notch intercellular domain (NICD) was observed in the nuclear fraction of GNAS wild-type cells. Meanwhile, inhibition of protein kinase A (PKA) induced NICD in GNAS-mutant IPMN cells, suggesting that NOTCH signaling is negatively regulated by the GNAS-PKA pathway. GNAS wild-type cells were characterized by a significant invasive property relative to GNAS-mutant cells, which was mediated through the NOTCH regulatory pathway.
Conclusions
Oncogenic GNAS induces mucin production, not only via MUC2 but also via MUC5AC/B, which may enlarge cystic lesions in the pancreas. The mutation may also limit tumor aggressiveness by attenuating NOTCH signaling; therefore, such tumor-suppressing effects must be considered when therapeutically inhibiting the GNAS pathway.
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Acknowledgements
We would like to thank Rika Kakisaka and Atsuko Nishikawa (Sapporo Higashi Tokushukai Hospital) for performing IHC quantification of the xenografts; Ayumu Sugitani (Sapporo Higashi Tokushukai Hospital) for supporting statistical analyses; and Eiko Aoyanagi (Sapporo Higashi Tokushukai Hospital) for tissue sample preparation. We also thank Krushna C. Patra (Department of Cancer Biology at University of Cincinnati) for critical reading of the manuscript.
Funding
This work was supported by Pancreas Research Foundation of Japan and Grants-in-Aid for Regional R&D Proposal-Based Program from Northern Advancement Center for Science & Technology of Hokkaido in Japan (to H. Kawabata) and by JSPS KAKENHI Grant Number 20K17009 (to H. Kawabata), 20K07671 (to Y.O.), 20K09070 (to H. Karasaki) and 20H03655 (to Y.M.).
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YO and YM receive funding from Hitachi High-Tech Corporation. The other authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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Kawabata, H., Ono, Y., Tamamura, N. et al. Mutant GNAS limits tumor aggressiveness in established pancreatic cancer via antagonizing the KRAS-pathway. J Gastroenterol 57, 208–220 (2022). https://doi.org/10.1007/s00535-021-01846-4
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DOI: https://doi.org/10.1007/s00535-021-01846-4