Identifying and targeting ROS1 gene fusions in non-small cell lung cancer.
Identifying and targeting ROS1 gene fusions in non-small cell lung cancer.
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DOI：
10.1158/1078-0432.ccr-12-0550
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发表时间：
2012-09-01
期刊：
影响因子：
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通讯作者：
Doebele RC
中科院分区：
文献类型：
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作者：
Davies KD;Le AT;Theodoro MF;Skokan MC;Aisner DL;Berge EM;Terracciano LM;Cappuzzo F;Incarbone M;Roncalli M;Alloisio M;Santoro A;Camidge DR;Varella-Garcia M;Doebele RC
已经在各种人类癌症中鉴定了涉及ROS 1激酶3'区的致癌基因融合。在这项研究中，我们试图在非小细胞肺癌（NSCLC）的特征ROS 1融合基因，并建立融合蛋白作为药物靶点。通过荧光原位杂交（FISH）对包含447个样本的NSCLC组织微阵列（TMA）面板进行ROS 1重排筛选。该检测也用于筛查NSCLC患者。对阳性样品进行反向PCR和RT-PCR鉴定。此外，通过用克唑替尼治疗ROS 1阳性患者，评估了ROS 1抑制的临床效用。表达SLC 34 A2-ROS 1融合的HCC 78细胞系用具有抗ROS 1活性的激酶抑制剂处理。通过MTS测定、流式细胞术和蛋白质印迹分析ROS 1抑制对增殖、细胞周期进展和细胞信号传导途径的影响。在TMA组中，发现5/428（1.2%）份可评价样本的ROS 1重排呈阳性。此外，1/48例患者重排检测呈阳性，该患者在克唑替尼治疗后显示肿瘤缩小。患者和一个TMA样品显示最近鉴定的SDC 4-ROS 1融合体的表达，而两个TMA样品表达CD 74-ROS 1融合体，另外两个表达SLC 34 A2-ROS 1融合体。在HCC 78细胞中，使用ROS 1抑制剂的治疗是抗增殖和下调信号通路，这对生长和存活至关重要。对于表达ROS 1融合基因的NSCLC患者，ROS 1抑制可能是一种有效的治疗策略。
Oncogenic gene fusions involving the 3’ region of ROS1 kinase have been identified in various human cancers. In this study, we sought to characterize ROS1 fusion genes in non-small cell lung cancer (NSCLC) and establish the fusion proteins as drug targets. A NSCLC tissue microarray (TMA) panel containing 447 samples was screened for ROS1 rearrangement by fluorescence in-situ hybridization (FISH). This assay was also used to screen NSCLC patients. In positive samples, the identity of the fusion partner was determined through inverse-PCR and RT-PCR. In addition, the clinical utility of ROS1 inhibition was assessed by treating a ROS1-positive patient with crizotinib. The HCC78 cell line, which expresses the SLC34A2-ROS1 fusion, was treated with kinase inhibitors that have activity against ROS1. The effects of ROS1 inhibition on proliferation, cell-cycle progression, and cell signaling pathways were analyzed by MTS assay, flow cytometry, and western blotting. In the TMA panel, 5/428 (1.2%) evaluable samples were found to be positive for ROS1 rearrangement. Additionally, 1/48 patients tested positive for rearrangement, and this patient demonstrated tumor shrinkage upon treatment with crizotinib. The patient and one TMA sample displayed expression of the recently identified SDC4-ROS1 fusion, while two TMA samples expressed the CD74-ROS1 fusion and two others expressed the SLC34A2-ROS1 fusion. In HCC78 cells, treatment with ROS1 inhibitors was anti-proliferative and down-regulated signaling pathways that are critical for growth and survival. ROS1 inhibition may be an effective treatment strategy for the subset of NSCLC patients whose tumors express ROS1 fusion genes.