MassIVE MSV000084223

Partial Public PXD015120

Loss of HIF1A From Pancreatic Cancer Cells Increases Expression of PPP1R1B and Degradation of p53 to Promote Invasion and Metastasis

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Description

Here we utilized a mouse model of PDAC and human pancreatic cancer cell lines to investigate the role of HIF1alpha in pancreatic cancer. Contrary to the conventional notion that HIF1alpha plays a tumor promoting role in several cancers, our findings indicate a tumor suppressive role of HIF1alpha in PDAC. Ablation of Hif1alpha in mice resulted in drastically increased metastasis of the pancreatic cancer and substantially reduced survival. We traced HIF1alpha's metastasis-suppression to the regulation of Protein Phosphatase 1 Regulatory Inhibitor subunit 1B (PPP1R1B), a dual kinase/phosphatase inhibited by HIF1alpha. We show a mechanistic link between loss of HIF1alpha, increased expression of PPP1R1B and degradation of the p53 protein that promotes invasion and metastasis in both mouse and human PDACs. Increased expression of PPP1R1B correlates with higher metastasis and poor survival in human pancreatic cancer patients. Since the inhibition of PPP1R1B reduced the metastatic potential of pancreatic cancer cells, we propose PPP1R1B as a therapeutic target to improve the prognosis of pancreatic cancer patients. _____ LC-MS/MS methodology _____ Dried peptide samples were reconstituted with 2% ACN-0.1% FA and quantified by NanoDropTM spectrophometer (ThermoFisher) prior to LC-MS/MS analysis using a nanoACQUITY system (Waters) coupled to an Orbitrap Fusion Lumos mass spectrometer (Thermo Fisher Scientific). Peptides were separated using a 200 cm micro-chip pillar array C18 column (PharmaFluidics) at a flow rate of 500 nl/min using a 123-min gradient: 1% to 5% B in 1 min, 5% to 10% B in 22 min, 10% to 18% B in 53 min, 18% to 28% B in 39 min, and 28% to 38% B in 8 min (A= FA 0.1%; B=100% ACN: 0.1% FA). The mass spectrometer was operated in positive data-dependent acquisition mode. MS1 spectra were measured in the Orbitrap with a resolution of 60000 (AGC target: 4e5; maximum injection time:50 ms; mass range: from 375 to 1500 m/z). The instrument was set to run in top speed mode with 2 s cycles for the survey and the MS/MS scans. After a survey scan, the most abundant precursors (with charge state between +2 and +7) were isolated in the quadrupole (isolation window: 1.6 m/z) and fragmented in the Ion Routing Multipole HCD-Cell (collision energy: 30%). fragmented precursors were detected in the ion trap cell with a rapid scan (First mass: 110 m/z; AGC target for MS/MS: 1e4; maximum injection time: 35 ms). The dynamic exclusion was set to 60 s with a 10 ppm mass tolerance around the precursor. [doi:10.25345/C5PM2P] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Pancreatic Ductal Adenocarcinoma (PDAC) ; Protein Phosphatase 1 Regulatory Inhibitor subunit 1B (PPP1R1B) ; metastasis ; HIF1 alpha

Contact

Principal Investigators:
(in alphabetical order) 		Anindya Bagchi, Sanford Burnham Prebys Medical Discovery Institute, United States
Submitting User: alexcampos

Publications

Tiwari A, Tashiro K, Dixit A, Soni A, Vogel K, Hall B, Shafqat I, Slaughter J, Param N, Le A, Saunders E, Paithane U, Garcia G, Campos AR, Zettervall J, Carlson M, Starr TK, Marahrens Y, Deshpande AJ, Commisso C, Provenzano PP, Bagchi A.
Loss of HIF1A From Pancreatic Cancer Cells Increases Expression of PPP1R1B and Degradation of p53 to Promote Invasion and Metastasis.
Gastroenterology. 2020 Nov;159(5):1882-1897.e5. Epub 2020 Aug 5.

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