Lampis Andrea, Carotenuto Pietro, Vlachogiannis Georgios, Cascione Luciano, Hedayat Somaieh, Burke Rosemary, Clarke Paul, Bosma Else, Simbolo Michele, Scarpa Aldo, Yu Sijia, Cole Rebecca, Smyth Elizabeth, Mateos Javier Fernández, Begum Ruwaida, Hezelova Blanka, Eltahir Zakaria, Wotherspoon Andrew, Fotiadis Nicos, Bali Maria Antonietta, Nepal Chirag, Khan Khurum, Stubbs Mark, Hahne Jens C, Gasparini Pierluigi, Guzzardo Vincenza, Croce Carlo M, Eccles Suzanne, Fassan Matteo, Cunningham David, Andersen Jesper B, Workman Paul, Valeri Nicola, Braconi Chiara
The Institute of Cancer Research, London, UK.
Bioinformatics Core Unit, Institute of Oncology Research, Bellinzona, Switzerland.
Gastroenterology. 2018 Mar;154(4):1066-1079.e5. doi: 10.1053/j.gastro.2017.10.043. Epub 2017 Nov 4.
BACKGROUND & AIMS: Cholangiocarcinomas (CCA) are resistant to chemotherapy, so new therapeutic agents are needed. We performed a screen to identify small-molecule compounds that are active against CCAs. Levels of microRNA 21 (MIR21 or miRNA21) are increased in CCAs. We investigated whether miRNA21 mediates resistance of CCA cells and organoids to HSP90 inhibitors.
We performed a high-throughput screen of 484 small-molecule compounds to identify those that reduced viability of 6 human CCA cell lines. We tested the effects of HSP90 inhibitors on cells with disruption of the MIR21 gene, cells incubated with MIR21 inhibitors, and stable cell lines with inducible expression of MIR21. We obtained CCA biopsies from patients, cultured them as organoids (patient-derived organoids). We assessed their architecture, mutation and gene expression patterns, response to compounds in culture, and when grown as subcutaneous xenograft tumors in mice.
Cells with IDH1 and PBRM1 mutations had the highest level of sensitivity to histone deacetylase inhibitors. HSP90 inhibitors were effective in all cell lines, irrespective of mutations. Sensitivity of cells to HSP90 inhibitors correlated inversely with baseline level of MIR21. Disruption of MIR21 increased cell sensitivity to HSP90 inhibitors. CCA cells that expressed transgenic MIR21 were more resistant to HSP90 inhibitors than cells transfected with control vectors; inactivation of MIR21 in these cells restored sensitivity to these agents. MIR21 was shown to target the DnaJ heat shock protein family (Hsp40) member B5 (DNAJB5). Transgenic expression of DNAJB5 in CCA cells that overexpressed MIR21 re-sensitized them to HSP90 inhibitors. Sensitivity of patient-derived organoids to HSP90 inhibitors, in culture and when grown as xenograft tumors in mice, depended on expression of miRNA21.
miRNA21 appears to mediate resistance of CCA cells to HSP90 inhibitors by reducing levels of DNAJB5. HSP90 inhibitors might be developed for the treatment of CCA and miRNA21 might be a marker of sensitivity to these agents.
胆管癌（CCA）对化疗耐药，因此需要新的治疗药物。我们进行了一项筛选，以鉴定对CCA有活性的小分子化合物。CCA中微小RNA 21（MIR21或miRNA21）水平升高。我们研究了miRNA21是否介导CCA细胞和类器官对HSP90抑制剂的耐药性。
我们对484种小分子化合物进行了高通量筛选，以鉴定那些能降低6种人CCA细胞系活力的化合物。我们测试了HSP90抑制剂对MIR21基因破坏的细胞、用MIR21抑制剂孵育的细胞以及具有MIR21诱导表达的稳定细胞系的影响。我们从患者身上获取CCA活检组织，将其培养成类器官（患者来源的类器官）。我们评估了它们的结构、突变和基因表达模式、对培养物中化合物的反应，以及在小鼠体内作为皮下异种移植肿瘤生长时的情况。
具有异柠檬酸脱氢酶1（IDH1）和溴结构域包含蛋白1（PBRM1）突变的细胞对组蛋白去乙酰化酶抑制剂的敏感性最高。HSP90抑制剂在所有细胞系中均有效，与突变无关。细胞对HSP90抑制剂的敏感性与MIR21的基线水平呈负相关。MIR21的破坏增加了细胞对HSP90抑制剂的敏感性。表达转基因MIR21的CCA细胞比用对照载体转染的细胞对HSP90抑制剂更耐药；这些细胞中MIR21的失活恢复了对这些药物的敏感性。已证明MIR21靶向DnaJ热休克蛋白家族（Hsp40）成员B5（DNAJB5）。在过表达MIR21的CCA细胞中DNAJB5的转基因表达使它们对HSP90抑制剂重新敏感。患者来源的类器官在培养物中以及在小鼠体内作为异种移植肿瘤生长时对HSP90抑制剂的敏感性取决于miRNA21的表达。
miRNA21似乎通过降低DNAJB5水平介导CCA细胞对HSP90抑制剂的耐药性。HSP90抑制剂可能被开发用于治疗CCA，而miRNA21可能是对这些药物敏感性的标志物。
