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Original Research Full Report: Basic and Translational—Pancreas| Volume 155, ISSUE 6, P1985-1998.e5, December 01, 2018

An Inhibitor of GSK3B and HDACs Kills Pancreatic Cancer Cells and Slows Pancreatic Tumor Growth and Metastasis in Mice

  • Mouad Edderkaoui
    Correspondence
    Reprint requests Address requests for reprints to: Mouad Edderkaoui, PhD, 8700 Beverly Boulevard, Davis 3100, Los Angeles, California 90048.
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California

    Department of Pediatrics, University of California at Los Angeles, Los Angeles, California
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  • Chintan Chheda
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • Badr Soufi
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • Fouzia Zayou
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • Robert W. Hu
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • V. Krishnan Ramanujan
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • Xinlei Pan
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • Laszlo G. Boros
    Affiliations
    Department of Pediatrics, University of California at Los Angeles, Los Angeles, California
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  • Jian Tajbakhsh
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • Anisha Madhav
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • Neil A. Bhowmick
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • Qiang Wang
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • Michael Lewis
    Affiliations
    Veterans Affairs, West Los Angeles, CA
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  • Richard Tuli
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • Aida Habtezion
    Affiliations
    Division of Gastroenterology and Hepatology, Department of Medicine, Stanford University School of Medicine, Stanford, California
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  • Ramachandran Murali
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California
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  • Stephen J. Pandol
    Affiliations
    Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California

    Department of Pediatrics, University of California at Los Angeles, Los Angeles, California
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      Background & Aims

      Growth, progression, and drug resistance of pancreatic ductal adenocarcinomas (PDACs) have been associated with increased levels and activity of glycogen synthase kinase 3 beta (GSK3B) and histone deacetylases (HDACs). We designed and synthesized molecules that simultaneously inhibit the activities of both enzymes. We tested the effects of one of these molecules, Metavert, in pancreatic cancer cells and mice with pancreatic tumors.

      Methods

      We tested the ability of Metavert to bind GSK3B and HDACs using surface plasmon resonance. MIA PaCa-2, Bx-PC3, HPAF-II, and HPDE6 cell lines were incubated with different concentrations of Metavert, with or without paclitaxel or gemcitabine, or with other inhibitors of GSK3B and HDACs; cells were analyzed for apoptosis and migration and by immunoblotting, immunofluorescence, and real-time polymerase chain reaction. Krasþ/LSLG12D;Trp53þ/LSLR172H;Pdx-1-Cre (KPC) mice (2 months old) were given injections of Metavert (5 mg/kg, 3 times/week) or vehicle (control). B6.129J mice with tumors grown from UN-KPC961-Luc cells were given injections of Metavert or vehicle. Tumors and metastases were counted and pancreata were analyzed by immunohistochemistry. Glucose metabolism was measured using 13C-glucose tracer and mass spectroscopy and flow cytometry. Cytokine levels in blood samples were measured using multiplexing enzyme-linked immunosorbent assay.

      Results

      Metavert significantly reduced survival of PDAC cells but not nontransformed cells; the agent reduced markers of the epithelial-to-mesenchymal transition and stem cells in PDAC cell lines. Cells incubated with Metavert in combination with irradiation and paclitaxel or gemcitabine had reduced survival compared with cells incubated with either agent alone; Metavert increased killing of drug-resistant PDAC cells by paclitaxel and gemcitabine. PDAC cells incubated with Metavert acquired normalized glucose metabolism. Administration of Metavert (alone or in combination with gemcitibine) to KPC mice or mice with syngeneic tumors significantly increased their survival times, slowed tumor growth, prevented tumor metastasis, decreased tumor infiltration by tumor-associated macrophages, and decreased blood levels of cytokines.

      Conclusions

      In studies of PDAC cells and 2 mouse models of PDAC, we found a dual inhibitor of GSK3B and HDACs (Metavert) to induce cancer cell apoptosis, reduce migration and expression of stem cell markers, and slow growth of tumors and metastases. Metavert had synergistic effects with gemcitabine.

      Keywords

      Abbreviations used in this paper:

      DAPI (4′,6-diamidino-2-phenylindole), EMT (epithelial-to-mesenchymal transition), GSK3B (glycogen synthase kinase 3 beta), HDAC (histone deacetylase), IL (interleukin), KPC (Krasþ/LSLG12D;Trp53þ/LSLR172H;Pdx-1-Cre), NF-κB (nuclear factor-κB), PDAC (pancreatic ductal adenocarcinomas), TCA (tricarboxylic acid)
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