Gene and microRNA modulation upon ET-743 treatment in a human intrahepatic cholangiocarcinoma paired patient derived xenograft and cell line [gene expression]. Homo sapiens

Transcriptional profiling and microRNA profiling of paired PDX and derived cell line MT-CHC01 upon ET-743 treatement Intrahepatic cholangiocarcinoma (ICC) is an aggressive and lethal malignancy with limited therapeutic options. ET-743 has a high antitumor activity in preclinical models of biliary tract carcinoma (BTC), being a promising alternative treatment. Here, we studied the effect of ET-743 at transcriptomic level on an ICC patient derived xenograft (PDX) and on the derived cell line, MT-CHC01. Further, putative targets of ET-743 were explored in the in vitro model. In vitro, ET-743 inhibited genes involved in protein modification, neurogenesis, migration, and motility; it induced the expression of genes involved in keratinization, tissues development, and apoptotic processes. In the PDX model, ET-743 affected ECM-receptor interaction, focal adhesion, complement and coagulation cascades, Hedgehog, MAPK, EGFR signaling via PIP3 pathway, and apoptosis. In MT-CHC01, 24 microRNAs were deregulated upon drug treatment. Only 5 microRNAs were perturbed by ET-743 in PDX; 2 up and 3 down-regulated. Among down- regulated genes, we selected SYK and LGALS1; their silencing caused a significantly reduction of migration, but did not affect proliferation in MT-CHC01 and WITT cells. In conclusion, we described that ET-743 affected genes and microRNAs involved in tumor progression and metastatic processes, reflecting data previously obtained at macroscopically level; in particular, we identified SYK and LGALS1 as new putative targets of ET-743. Overall design: MT-CHC01 cell line treated with ET-743 at the dose of 5 nM for 24 hours vs MT-CHC01 cell line untreated;ICC PDX treated with ET-743 at the dose of 0.15 mg/Kg/weekly for three weeks vs ICC PDX untreated

ET-743处理后配对的患者来源异种移植瘤模型（Patient Derived Xenograft, PDX）及其来源细胞系MT-CHC01的转录组谱分析与微小RNA（microRNA）谱分析

肝内胆管癌（Intrahepatic Cholangiocarcinoma, ICC）是一种侵袭性强、致死性高的恶性肿瘤，临床治疗选择极为有限。ET-743在胆道癌（Biliary Tract Carcinoma, BTC）的临床前模型中展现出优异的抗肿瘤活性，是极具潜力的替代治疗方案。本研究从转录组层面探究了ET-743对ICC患者来源异种移植瘤模型及其来源细胞系MT-CHC01的作用，并进一步在体外模型中探索了ET-743的潜在作用靶点。

体外实验中，ET-743可抑制参与蛋白质修饰、神经发生、细胞迁移与运动的基因表达，并上调参与角质形成、组织发育及凋亡过程的基因转录水平。在PDX模型中，ET-743可调控细胞外基质-受体相互作用、黏着斑、补体与凝血级联反应、刺猬（Hedgehog）信号通路、丝裂原活化蛋白激酶（MAPK）通路、经磷脂酰肌醇三磷酸（PIP3）途径的表皮生长因子受体（EGFR）信号通路以及细胞凋亡相关过程。

在MT-CHC01细胞系中，药物处理后共有24个微小RNA表达失调；而在PDX模型中，仅5个微小RNA受ET-743调控，其中2个上调、3个下调。在下调基因中，我们筛选出SYK与LGALS1；敲低这两个基因可显著抑制MT-CHC01与WITT细胞的迁移能力，但对细胞增殖无明显影响。

综上，本研究证实ET-743可调控参与肿瘤进展与转移过程的基因及微小RNA表达，与此前宏观层面获得的研究结果一致；尤为重要的是，本研究鉴定出SYK与LGALS1为ET-743的新型潜在作用靶点。

实验设计：以5 nM浓度的ET-743处理MT-CHC01细胞系24小时，以未处理的MT-CHC01细胞系作为对照；以0.15 mg/kg/周的ET-743剂量处理ICC PDX模型三周，以未处理的ICC PDX模型作为对照。