Huijie Miao1, Yajun Geng1, Yang Li1, Shijie Tang2,3, Feiling Feng4, Weijian Li1, Yongsheng Li1, Liguo Liu1, Rui Zhang5, Shimei Qiu6,7, Ying Wu6,7, Zeyu Wang1, Ziyi Wang1, Ziyu Shao8, Ke Liu1, Lu Zou1, Mao Yang1, Yuhao Zhao1, Chen Chen6,7, Zhizhen Li4, Dadong Zhang9, Peng Peng10, Xiaoyan Qiang10, Frank Wu10, Yongning He6, Luonan Chen2,11,12, Dongxi Xiang13, Xiaoqing Jiang14, Maolan Li15,16, Yun Liu17, Yingbin Liu18,19,20. 1. Department of Biliary-Pancreatic Surgery, Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China. 2. State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, CAS Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai, 200031, China. 3. University of Chinese Academy of Sciences, Beijing, 100049, China. 4. Department of Biliary Surgery, Eastern Hepatobiliary Surgery Hospital, Shanghai, 200433, China. 5. Department of Hepatobiliary Surgery, the First Affiliated Hospital of Xi'an Jiao Tong University, Xi'an, 710061, China. 6. State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai, 200127, China. 7. Shanghai Key Laboratory of Biliary Tract Disease Research, Shanghai, 200127, China. 8. Department of General Surgery, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200092, China. 9. 3D Medicines Lnc, Shanghai, 201114, China. 10. Transthera Sciences (Nanjing), lnc, Nanjing, 210032, Jiangsu, China. 11. Center for Excellence in Animal Evolution and Genetics, Chinese Academy of Sciences, Kunming, 650223, China. 12. Key Laboratory of Systems Health Science of Zhejiang Province, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Hangzhou, 310024, China. 13. State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai, 200127, China. dxiang@shsmu.edu.cn. 14. Department of Biliary Surgery, Eastern Hepatobiliary Surgery Hospital, Shanghai, 200433, China. jxqehbh@sina.com. 15. Department of Biliary-Pancreatic Surgery, Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China. limaolan6@163.com. 16. Shanghai Key Laboratory of Biliary Tract Disease Research, Shanghai, 200127, China. limaolan6@163.com. 17. State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai, 200127, China. superliuyun@fudan.edu.cn. 18. Department of Biliary-Pancreatic Surgery, Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China. laoniulyb@shsmu.edu.cn. 19. State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai, 200127, China. laoniulyb@shsmu.edu.cn. 20. Shanghai Key Laboratory of Biliary Tract Disease Research, Shanghai, 200127, China. laoniulyb@shsmu.edu.cn.
Abstract
PURPOSE: This study aimed to investigate the efficiency of our chemically synthesized TT-00420, a novel spectrum-selective multiple protein kinase inhibitor, in cultured cells and animal models of gallbladder cancer (GBC) and explore its potential mechanism. METHODS: Multiple GBC models were established to assess the anti-tumor efficiency, toxicity, and pharmacokinetics of TT-00420. Integrated transcriptomic, proteomic and phosphoproteomic analysis was conducted to identify potential downstream effectors of TT-00420. Western blotting, qRT-PCR, nuclear-cytoplasm separation, and immunofluorescence were performed to confirm the multi-omic results and explore the molecular mechanism of TT-00420. Immunohistochemistry was used to detect FGFR1 and p-FGFR1 expression levels in GBC samples. Autodock software was utilized to investigate the potential binding mode between the TT-00420 and the human FGFR1. RESULTS: We found that TT-00420 exerted potent growth inhibition of GBC cell lines and multiple xenograft models. Treatment of mice with 15 mg/kg TT-00420 via gavage displayed a half-life of 1.8 h in the blood and rapid distribution to the liver, kidneys, lungs, spleen, and tumors at 0.25 h, but no toxicity to these organs over 2 weeks. Multi-omic analysis revealed c-Jun as a potential downstream effector after TT-00420 treatment. Mechanistically, TT-00420 showed rigorous ability to block FGFR1 and its downstream JNK-JUN (S63/S73) signaling pathway, and induce c-Jun S243-dependent MEK/ERK reactivation, leading to FASLG-dependent tumor cell death. Finally, we found that FGFR1 and p-FGFR1 expression was elevated in GBC patients and these levels correlated with decreased patient survival. CONCLUSIONS: TT-00420 shows potent antitumor efficacy and may serve as a novel agent to improve GBC prognosis.
PURPOSE: This study aimed to investigate the efficiency of our chemically synthesized TT-00420, a novel spectrum-selective multiple protein kinase inhibitor, in cultured cells and animal models of gallbladder cancer (GBC) and explore its potential mechanism. METHODS: Multiple GBC models were established to assess the anti-tumor efficiency, toxicity, and pharmacokinetics of TT-00420. Integrated transcriptomic, proteomic and phosphoproteomic analysis was conducted to identify potential downstream effectors of TT-00420. Western blotting, qRT-PCR, nuclear-cytoplasm separation, and immunofluorescence were performed to confirm the multi-omic results and explore the molecular mechanism of TT-00420. Immunohistochemistry was used to detect FGFR1 and p-FGFR1 expression levels in GBC samples. Autodock software was utilized to investigate the potential binding mode between the TT-00420 and the human FGFR1. RESULTS: We found that TT-00420 exerted potent growth inhibition of GBC cell lines and multiple xenograft models. Treatment of mice with 15 mg/kg TT-00420 via gavage displayed a half-life of 1.8 h in the blood and rapid distribution to the liver, kidneys, lungs, spleen, and tumors at 0.25 h, but no toxicity to these organs over 2 weeks. Multi-omic analysis revealed c-Jun as a potential downstream effector after TT-00420 treatment. Mechanistically, TT-00420 showed rigorous ability to block FGFR1 and its downstream JNK-JUN (S63/S73) signaling pathway, and induce c-Jun S243-dependent MEK/ERK reactivation, leading to FASLG-dependent tumor cell death. Finally, we found that FGFR1 and p-FGFR1 expression was elevated in GBC patients and these levels correlated with decreased patient survival. CONCLUSIONS: TT-00420 shows potent antitumor efficacy and may serve as a novel agent to improve GBC prognosis.
Authors: Rachna T Shroff; Milind M Javle; Lianchun Xiao; Ahmed O Kaseb; Gauri R Varadhachary; Robert A Wolff; Kanwal P S Raghav; Michiko Iwasaki; Peter Masci; Ramesh K Ramanathan; Daniel H Ahn; Tanios S Bekaii-Saab; Mitesh J Borad Journal: JAMA Oncol Date: 2019-06-01 Impact factor: 31.777