Krook Melanie A, Bonneville Russell, Chen Hui-Zi, Reeser Julie W, Wing Michele R, Martin Dorrelyn M, Smith Amy M, Dao Thuy, Samorodnitsky Eric, Paruchuri Anoosha, Miya Jharna, Baker Kaitlin R, Yu Lianbo, Timmers Cynthia, Dittmar Kristin, Freud Aharon G, Allenby Patricia, Roychowdhury Sameek
Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210, USA.
Biomedical Sciences Graduate Program, The Ohio State University, Columbus, Ohio 43210, USA.
Cold Spring Harb Mol Case Stud. 2019 Aug 1;5(4). doi: 10.1101/mcs.a004002. Print 2019 Aug.
Cholangiocarcinoma is a highly aggressive and lethal malignancy, with limited treatment options available. Recently, FGFR inhibitors have been developed and utilized in FGFR-mutant cholangiocarcinoma; however, resistance often develops and the genomic determinants of resistance are not fully characterized. We completed whole-exome sequencing (WES) of 11 unique tumor samples obtained from a rapid research autopsy on a patient with FGFR-fusion-positive cholangiocarcinoma who initially responded to the pan-FGFR inhibitor, INCB054828. In vitro studies were carried out to characterize the novel FGFR alteration and secondary mutation identified. Multisite WES and analysis of tumor heterogeneity through subclonal inference identified four genetically distinct cancer cell populations, two of which were only observed after treatment. Additionally, WES revealed an N549H mutation hypothesized to confer resistance to the FGFR inhibitor INCB054828 in a single tumor sample. This hypothesis was corroborated with in vitro cell-based studies in which cells expressing fusion were sensitive to INCB054828 (IC value of 10.16 nM), whereas cells with the addition of the N549H mutation were resistant to INCB054828 (IC value of 1527.57 nM). Furthermore, the N549H secondary mutation displayed cross-resistance to other selective FGFR inhibitors, but remained sensitive to the nonselective inhibitor, ponatinib. Rapid research autopsy has the potential to provide unprecedented insights into the clonal evolution of cancer throughout the course of the disease. In this study, we demonstrate the emergence of a drug resistance mutation and characterize the evolution of tumor subclones within a cholangiocarcinoma disease course.
胆管癌是一种侵袭性很强且致命的恶性肿瘤，可用的治疗选择有限。最近，FGFR抑制剂已被开发并应用于FGFR突变的胆管癌；然而，耐药性常常出现，且耐药的基因组决定因素尚未完全明确。我们对一名FGFR融合阳性胆管癌患者进行快速研究尸检时获得的11个独特肿瘤样本进行了全外显子组测序（WES），该患者最初对泛FGFR抑制剂INCB054828有反应。进行了体外研究以表征鉴定出的新型FGFR改变和二次突变。通过亚克隆推断进行的多位点WES和肿瘤异质性分析确定了四个基因上不同的癌细胞群体，其中两个仅在治疗后观察到。此外，WES在一个肿瘤样本中发现了一个N549H突变，推测该突变赋予对FGFR抑制剂INCB054828的耐药性。这一假设在基于细胞的体外研究中得到了证实，其中表达融合蛋白的细胞对INCB054828敏感（IC值为10.16 nM），而添加了N549H突变的细胞对INCB054828耐药（IC值为1527.57 nM）。此外，N549H二次突变对其他选择性FGFR抑制剂表现出交叉耐药性，但对非选择性抑制剂波纳替尼仍敏感。快速研究尸检有可能为癌症在整个疾病过程中的克隆进化提供前所未有的见解。在本研究中，我们展示了一种耐药性突变的出现，并表征了胆管癌疾病过程中肿瘤亚克隆的进化。
