Mutant GNAS limits tumor aggressiveness in established pancreatic cancer via antagonizing the KRAS-pathway
Mutant GNAS limits tumor aggressiveness in established pancreatic cancer via antagonizing the KRAS-pathway
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DOI：
10.1007/s00535-021-01846-4
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发表时间：
2022-01
影响因子：
6.3
通讯作者：
Hidemasa Kawabata;Y. Ono;Nobue Tamamura;Kyohei Oyama;J. Ueda;Hiroki Sato-;Kenji Takahashi;Kenzui Taniue;Tetsuhiro Okada;Syugo Fujibayashi;Akihiro Hayashi;T. Goto;Katsuro Enomoto;H. Konishi;M. Fujiya;Keita Miyakawa;M. Tanino;Y. Nishikawa;D. Koga;Tsuyoshi Watanabe;Chiho Maeda;H. Karasaki;A. Liss;Y. Mizukami;T. Okumura
中科院分区：
文献类型：
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作者：
Hidemasa Kawabata;Y. Ono;Nobue Tamamura;Kyohei Oyama;J. Ueda;Hiroki Sato-;Kenji Takahashi;Kenzui Taniue;Tetsuhiro Okada;Syugo Fujibayashi;Akihiro Hayashi;T. Goto;Katsuro Enomoto;H. Konishi;M. Fujiya;Keita Miyakawa;M. Tanino;Y. Nishikawa;D. Koga;Tsuyoshi Watanabe;Chiho Maeda;H. Karasaki;A. Liss;Y. Mizukami;T. Okumura
背景资料：GNAS突变驱动胰腺肿瘤发生，并经常发生在导管内乳头状粘液性肿瘤（IPMN）;然而，其作为治疗靶点的价值尚未确定。本研究旨在评估突变体GNAS在已建立的胰腺癌肿瘤侵袭性中的参与。方法：使用人原代IPMN相关胰腺癌细胞进行CRISPR/Cas9介导的GNAS R201 H沉默。通过进行细胞培养和异种移植实验，评价致癌GNAS在肿瘤维持中的作用，并进行蛋白质印迹和转录组分析，以揭示GNAS驱动的signatures.Results：GNAS野生型细胞的异种移植物的特征在于相对于GNAS突变细胞具有更高的Ki-67标记指数。GNAS野生型肿瘤中的表型改变导致粘蛋白产生显著减少，伴有大量基质成分的固体。转录谱分析表明突变GNAS与KRAS信号传导存在明显冲突。在GNAS野生型细胞的核部分中观察到显著更高的Notch细胞间结构域（NICD）。与此同时，蛋白激酶A（PKA）的抑制诱导了GNAS突变IPMN细胞中的NICD，表明NOTCH信号传导受到GNAS-PKA途径的负调节。GNAS野生型细胞的特点是相对于GNAS突变细胞，这是通过NOTCH调节pathway.Conclusions介导的一个显着的侵入性属性：致癌GNAS诱导粘蛋白的生产，不仅通过MUC 2，但也通过MUC 5AC/B，这可能会扩大胰腺囊性病变。该突变还可以通过减弱NOTCH信号传导来限制肿瘤的侵袭性;因此，在治疗性抑制GNAS途径时必须考虑这种肿瘤抑制作用。
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.