Mouad Edderkaoui1, Chintan Chheda2, Badr Soufi2, Fouzia Zayou2, Robert W Hu2, V Krishnan Ramanujan2, Xinlei Pan2, Laszlo G Boros3, Jian Tajbakhsh2, Anisha Madhav2, Neil A Bhowmick2, Qiang Wang2, Michael Lewis4, Richard Tuli2, Aida Habtezion5, Ramachandran Murali2, Stephen J Pandol6. 1. 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. Electronic address: mouad.edderkaoui@cshs.org. 2. Departments of Medicine, Biomedical Sciences, Radiation Oncology, and Surgery, Samuel Oschin Comprehensive Cancer Center, Cedars-Sinai Medical Center, Los Angeles, California. 3. Department of Pediatrics, University of California at Los Angeles, Los Angeles, California. 4. Veterans Affairs, West Los Angeles, CA. 5. Division of Gastroenterology and Hepatology, Department of Medicine, Stanford University School of Medicine, Stanford, California. 6. 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.
Abstract
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.
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.
Authors: B W Park; H T Zhang; C Wu; A Berezov; X Zhang; R Dua; Q Wang; G Kao; D M O'Rourke; M I Greene; R Murali Journal: Nat Biotechnol Date: 2000-02 Impact factor: 54.908
Authors: M S Finnin; J R Donigian; A Cohen; V M Richon; R A Rifkind; P A Marks; R Breslow; N P Pavletich Journal: Nature Date: 1999-09-09 Impact factor: 49.962
Authors: Andrei V Ougolkov; Martin E Fernandez-Zapico; Doris N Savoy; Raul A Urrutia; Daniel D Billadeau Journal: Cancer Res Date: 2005-03-15 Impact factor: 12.701
Authors: Sunil R Hingorani; Lifu Wang; Asha S Multani; Chelsea Combs; Therese B Deramaudt; Ralph H Hruban; Anil K Rustgi; Sandy Chang; David A Tuveson Journal: Cancer Cell Date: 2005-05 Impact factor: 31.743
Authors: Mouad Edderkaoui; Peggy Hong; Eva C Vaquero; Jong K Lee; Lars Fischer; Helmut Friess; Markus W Buchler; Markus M Lerch; Stephen J Pandol; Anna S Gukovskaya Journal: Am J Physiol Gastrointest Liver Physiol Date: 2005-07-21 Impact factor: 4.052
Authors: Sunil R Hingorani; Emanuel F Petricoin; Anirban Maitra; Vinodh Rajapakse; Catrina King; Michael A Jacobetz; Sally Ross; Thomas P Conrads; Timothy D Veenstra; Ben A Hitt; Yoshiya Kawaguchi; Don Johann; Lance A Liotta; Howard C Crawford; Mary E Putt; Tyler Jacks; Christopher V E Wright; Ralph H Hruban; Andrew M Lowy; David A Tuveson Journal: Cancer Cell Date: 2003-12 Impact factor: 31.743
Authors: Robin E Bachelder; Sang-Oh Yoon; Clara Franci; Antonio García de Herreros; Arthur M Mercurio Journal: J Cell Biol Date: 2005-01-03 Impact factor: 10.539
Authors: Vera Mayo; Annie C Bowles; Laura E Wubker; Ismael Ortiz; Albert M Cordoves; Richard J Cote; Diego Correa; Ashutosh Agarwal Journal: Exp Biol Med (Maywood) Date: 2020-11-19