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Loss of the wild-type KRAS allele promotes pancreatic cancer progression through functional activation of YAP1

Abstract

Human pancreatic ductal adenocarcinoma (PDAC) harboring one KRAS mutant allele often displays increasing genomic loss of the remaining wild-type (WT) allele (known as LOH at KRAS) as tumors progress to metastasis, yet the molecular ramification of this WT allelic loss is unknown. In this study, we showed that the restoration of WT KRAS expression in human PDAC cell lines with LOH at KRAS significantly attenuated the malignancy of PDAC cells both in vitro and in vivo, demonstrating a tumor-suppressive role of the WT KRAS allele. Through RNA-Seq, we identified the HIPPO signaling pathway to be positively regulated by WT KRAS in PDAC cells. In accordance with this observation, PDAC cells with LOH at KRAS exhibited increased nuclear localization and activation of transcriptional co-activator YAP1. Mechanistically, we discovered that WT KRAS expression sequestered YAP1 from the nucleus, through enhanced 14-3-3zeta interaction with phosphorylated YAP1 at S127. Consistently, expression of a constitutively-active YAP1 mutant in PDAC cells bypassed the growth inhibitory effects of WT KRAS. In patient samples, we found that the YAP1-activation genes were significantly upregulated in tumors with LOH at KRAS, and YAP1 nuclear localization predicted poor survival for PDAC patients. Collectively, our results reveal that the WT allelic loss leads to functional activation of YAP1 and enhanced tumor malignancy, which explains the selection advantage of the tumor cells with LOH at KRAS during pancreatic cancer clonal evolution and progression to metastasis, and should be taken into consideration in future therapeutic strategies targeting KRAS.

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Fig. 1: Restored wild-type KRAS expression in pancreatic cancer cell lines attenuated tumor malignancies in vitro and in vivo.
Fig. 2: Induced WT KRAS expression attenuated YAP1 activation in the iWT-KRAS MIA PaCa-2 and iWT-KRAS AsPC-1 cell lines.
Fig. 3: The restoration of WT KRAS expression led to enhanced YAP1-14-3-3zeta binding and reduced YAP1-TEAD activities.
Fig. 4: The S127A mutant YAP1 reversed the tumor-suppression exerted by the wild-type KRAS allele in the iWT-KRAS-Mia-PaCa-2 cells.
Fig. 5: LOH at KRAS was positively associated with the increased expression of the YAP1-activation signature in human PDAC samples.

Data availability

RNA-Seq data have been deposited in the Gene Expression Omnibus (GEO) with Accession GSE156562. Proteomics data have been uploaded to the MassIVE data repository site with the accession ID: MassIVE MSV000088107.

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Acknowledgements

This study was supported by NIH/NCI R01 CA178445, NIH/NCI R01 CA217207, CUIMC Irving Drug Discovery Pilot Award (HICCC_FY21IDD01), and NIH P30CA013696 for the HICCC. HYan was supported by the China Scholarship Council for research at the CUIMC. YY was supported by the Morgan Stanley Children’s Hospital-Beijing Children’s Hospital Program. DCK was supported by the NCI-ICBP (Integrative Cancer Biology Program) Summer Research Program.

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HYan and CY were responsible for the designs of the experiments, the acquisition, analyses, interpretation, and presentation of the data, and drafting the paper. SAF, ALY, YY, JY, DCK, ECC, EIC, and WQ contributed to the acquisition and analyses of the data. RAF, HYing, and EIC were responsible for the computational analyses, statistical analyses, and interpretation of the data. EIC, JL, YM, and GHS provided resources and materials that were vital to the completion of the work. HYan, CY, SAF, RAF, EIC, JL, WQ, and GHS prepared and revised the paper. WQ and GHS conceived and designed the work that led to the submission. Each author has made substantial contributions to the work and/or have been involved in drafting or revising the paper. All the authors have given final approval of the version to be published and take public responsibility for appropriate portions of the content.

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Correspondence to Gloria H. Su.

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Yan, H., Yu, CC., Fine, S.A. et al. Loss of the wild-type KRAS allele promotes pancreatic cancer progression through functional activation of YAP1. Oncogene 40, 6759–6771 (2021). https://doi.org/10.1038/s41388-021-02040-9

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