Department of Biliary-Pancreatic Surgery, Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China.
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.
Cell Oncol (Dordr). 2022 Aug;45(4):689-708. doi: 10.1007/s13402-022-00692-7. Epub 2022 Jul 23.
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.
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.
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.
TT-00420 shows potent antitumor efficacy and may serve as a novel agent to improve GBC prognosis.
本研究旨在探究我们合成的新型谱选择性多蛋白激酶抑制剂 TT-00420 在胆囊癌（GBC）培养细胞和动物模型中的疗效，并探索其潜在机制。
建立多种 GBC 模型，评估 TT-00420 的抗肿瘤效率、毒性和药代动力学。进行整合转录组学、蛋白质组学和磷酸化蛋白质组学分析，以鉴定 TT-00420 的潜在下游效应物。通过 Western blot、qRT-PCR、核质分离和免疫荧光实验，对多组学结果进行验证，并探讨 TT-00420 的分子机制。免疫组化检测 GBC 样本中 FGFR1 和 p-FGFR1 的表达水平。利用 Autodock 软件研究 TT-00420 与人类 FGFR1 之间的潜在结合模式。
我们发现 TT-00420 对 GBC 细胞系和多种异种移植模型均具有强大的生长抑制作用。灌胃给予小鼠 15mg/kg TT-00420 后，其在血液中的半衰期为 1.8 小时，在 0.25 小时时迅速分布到肝脏、肾脏、肺、脾脏和肿瘤中，但在 2 周内对这些器官没有毒性。多组学分析表明，TT-00420 处理后 c-Jun 是潜在的下游效应物。在机制上，TT-00420 表现出严格阻断 FGFR1 及其下游 JNK-JUN（S63/S73）信号通路的能力，并诱导 c-Jun S243 依赖性 MEK/ERK 重新激活，导致 FASLG 依赖性肿瘤细胞死亡。最后，我们发现 FGFR1 和 p-FGFR1 的表达在 GBC 患者中升高，这些水平与患者生存时间缩短相关。
TT-00420 显示出强大的抗肿瘤疗效，可能成为改善 GBC 预后的新型药物。
