Novel protein kinase inhibitor TT-00420 inhibits gallbladder cancer by inhibiting JNK/JUN-mediated signaling pathway

  • Cell Oncol (Dordr). 2022 Aug;45(4):689-708. doi: 10.1007/s13402-022-00692-7.
Huijie Miao  #  1 ,  Yajun Geng  #  1 ,  Yang Li  #  1 ,  Shijie Tang  #  2  3 ,  Feiling Feng  #  4 ,  Weijian Li  1 ,  Yongsheng Li  1 ,  Liguo Liu  1 ,  Rui Zhang  5 ,  Shimei Qiu  6  7 ,  Ying Wu  6  7 ,  Zeyu Wang  1 ,  Ziyi Wang  1 ,  Ziyu Shao  8 ,  Ke Liu  1 ,  Lu Zou  1 ,  Mao Yang  1 ,  Yuhao Zhao  1 ,  Chen Chen  6  7 ,  Zhizhen Li  4 ,  Dadong Zhang  9 ,  Peng Peng  10 ,  Xiaoyan Qiang  10 ,  Frank Wu  10 ,  Yongning He  6 ,  Luonan Chen  2  11  12 ,  Dongxi Xiang  13 ,  Xiaoqing Jiang  14 ,  Maolan Li  15  16 ,  Yun Liu  17 ,  Yingbin Liu  18  19  20
Affiliations
  • 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. [email protected].
  • 14. Department of Biliary Surgery, Eastern Hepatobiliary Surgery Hospital, Shanghai, 200433, China. [email protected].
  • 15. Department of Biliary-Pancreatic Surgery, Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China. [email protected].
  • 16. Shanghai Key Laboratory of Biliary Tract Disease Research, Shanghai, 200127, China. [email protected].
  • 17. State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai, 200127, China. [email protected].
  • 18. Department of Biliary-Pancreatic Surgery, Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China. [email protected].
  • 19. State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai, 200127, China. [email protected].
  • 20. Shanghai Key Laboratory of Biliary Tract Disease Research, Shanghai, 200127, China. [email protected].
  • # Contributed equally.
PMID: 35870050 DOI: 10.1007/s13402-022-00692-7
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.

Keywords
Cell Apoptosis; Gallbladder cancer; Multiple kinase inhibitor; Phosphoproteomics; Proteomics.
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