Comparative Proteomic Analysis Identifies Key Metabolic Regulators of Gemcitabine Resistance in Pancreatic Cancer

Qingxiang Lin; Shichen Shen; Zhicheng Qian; Sailee S Rasam; Andrea Serratore; William J Jusko; Eugene S Kandel; Jun Qu; Robert M Straubinger

doi:10.1016/j.mcpro.2022.100409

Comparative Proteomic Analysis Identifies Key Metabolic Regulators of Gemcitabine Resistance in Pancreatic Cancer

Mol Cell Proteomics. 2022 Oct;21(10):100409. doi: 10.1016/j.mcpro.2022.100409. Epub 2022 Sep 7.

Authors

Qingxiang Lin¹, Shichen Shen², Zhicheng Qian³, Sailee S Rasam⁴, Andrea Serratore³, William J Jusko³, Eugene S Kandel⁵, Jun Qu⁶, Robert M Straubinger⁷

Affiliations

¹ Department of Cell Stress Biology, Roswell Park Comprehensive Cancer Center, Buffalo, New York, USA; Department of Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, New York, USA; Center of Excellence in Bioinformatics & Life Science, University at Buffalo, State University of New York, Buffalo, New York, USA.
² Department of Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, New York, USA; Center of Excellence in Bioinformatics & Life Science, University at Buffalo, State University of New York, Buffalo, New York, USA.
³ Department of Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, New York, USA.
⁴ Center of Excellence in Bioinformatics & Life Science, University at Buffalo, State University of New York, Buffalo, New York, USA; Department of Biochemistry, University at Buffalo, State University of New York, Buffalo, New York, USA.
⁵ Department of Cell Stress Biology, Roswell Park Comprehensive Cancer Center, Buffalo, New York, USA.
⁶ Department of Cell Stress Biology, Roswell Park Comprehensive Cancer Center, Buffalo, New York, USA; Department of Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, New York, USA; Center of Excellence in Bioinformatics & Life Science, University at Buffalo, State University of New York, Buffalo, New York, USA; Department of Biochemistry, University at Buffalo, State University of New York, Buffalo, New York, USA. Electronic address: junqu@buffalo.edu.
⁷ Department of Cell Stress Biology, Roswell Park Comprehensive Cancer Center, Buffalo, New York, USA; Department of Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, New York, USA; Center of Excellence in Bioinformatics & Life Science, University at Buffalo, State University of New York, Buffalo, New York, USA; Department of Pharmacology & Therapeutics, Roswell Park Comprehensive Cancer Center, Buffalo, New York, USA. Electronic address: rms@buffalo.edu.

Abstract

Pancreatic adenocarcinoma (PDAC) is highly refractory to treatment. Standard-of-care gemcitabine (Gem) provides only modest survival benefits, and development of Gem resistance (GemR) compromises its efficacy. Highly GemR clones of Gem-sensitive MIAPaCa-2 cells were developed to investigate the molecular mechanisms of GemR and implemented global quantitative differential proteomics analysis with a comprehensive, reproducible ion-current-based MS1 workflow to quantify ∼6000 proteins in all samples. In GemR clone MIA-GR8, cellular metabolism, proliferation, migration, and 'drug response' mechanisms were the predominant biological processes altered, consistent with cell phenotypic alterations in cell cycle and motility. S100 calcium binding protein A4 was the most downregulated protein, as were proteins associated with glycolytic and oxidative energy production. Both responses would reduce tumor proliferation. Upregulation of mesenchymal markers was prominent, and cellular invasiveness increased. Key enzymes in Gem metabolism pathways were altered such that intracellular utilization of Gem would decrease. Ribonucleoside-diphosphate reductase large subunit was the most elevated Gem metabolizing protein, supporting its critical role in GemR. Lower Ribonucleoside-diphosphate reductase large subunit expression is associated with better clinical outcomes in PDAC, and its downregulation paralleled reduced MIAPaCa-2 proliferation and migration and increased Gem sensitivity. Temporal protein-level Gem responses of MIAPaCa-2 versus GemR cell lines (intrinsically GemR PANC-1 and acquired GemR MIA-GR8) implicate adaptive changes in cellular response systems for cell proliferation and drug transport and metabolism, which reduce cytotoxic Gem metabolites, in DNA repair, and additional responses, as key contributors to the complexity of GemR in PDAC. These findings additionally suggest targetable therapeutic vulnerabilities for GemR PDAC patients.

Keywords: comparative proteomics; drug metabolism; gemcitabine resistance; pancreatic cancer.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Adenocarcinoma*
Cell Line, Tumor
Diphosphates / metabolism
Diphosphates / therapeutic use
Drug Resistance, Neoplasm / genetics
Gemcitabine
Humans
Pancreatic Neoplasms* / metabolism
Proteomics
Ribonucleosides* / therapeutic use
S100 Calcium-Binding Protein A4

Substances

Diphosphates
Ribonucleosides
S100 Calcium-Binding Protein A4
Gemcitabine

Abstract

Publication types

MeSH terms

Substances

Grants and funding