2012年12月14日訊 /生物谷BIOON/ --大約20例非小細胞肺癌（NSCLC）患者中有一例間變性淋巴瘤激酶（ALK）基因染色體畸變,，這些患者對ALK-激酶抑制劑如Crizotinib治療高度敏感,。然而，目前的診斷測試有其局限性,。
研究人員已經開發(fā)并測試了一種有前途的新方法篩選非小細胞肺癌的ALK融合,。新研究提供了一個成本效益和易于執(zhí)行的新測試方法。這項研究發(fā)表在Molecular Diagnostics雜志上。
Crizotinib是一種蛋白酪氨酸激酶抑制劑,，被FDA批準用于治療ALK陽性的局部晚期或轉移性非小細胞肺癌患者,，目前正在進行Ⅲ期臨床試驗研究。
確定哪些患者最有可能受益于ALK抑制是非常重要的?，F(xiàn)在最新的國家綜合癌癥網(wǎng)絡（NCCN）臨床腫瘤學臨床實踐指南建議對所有非小細胞肺癌患者進行前期ALK篩選,。
有幾種經臨床驗證的方法目前可用于檢測ALK融合，包括熒光原位雜交（FISH）,，免疫組化法（IHC）和逆轉錄聚合酶鏈反應（RT-PCR）,。目前利用熒光原位雜交為基礎的測試最近被FDA批準為Crizotinib的標準診斷測試。
然而,，熒光原位雜交為基礎的測試是復雜的,，成本高,。有相當大的限制使得難以篩選大量患者,。但FISH檢測在臨床上得到了廣泛的驗證，是目前檢測ALK融合的黃金標準,。
但是這一診斷方法的缺點在于檢測所得信號是微妙的,，同時需要專門的技術知識來解釋。與IHC和RT-PCR相比也相當昂貴,。
要探索替代篩選檢測ALK融合的現(xiàn)有方式,，他們設計了一種新的方法，通過使用基因表達平臺直接檢測ALK融合,。他們利用66例非小細胞肺癌樣品測試了他們的新方法,，其結果與之前FISH和IHC的結果一致。
利用新方法,，研究人員能成功地利用少量腫瘤細胞樣品檢測到ALK融合基因,。雖然進一步的測試還需要一個更大的樣本量，但我們已經證明新方法是一個具有成本效益,，易于操作,，高通量的篩選檢測ALK融合的方式。（生物谷Bioon.com）
doi: 10.1016/j.jmoldx.2012.08.006
Multiplexed Gene Expression and Fusion Transcript Analysis to Detect ALK Fusions in Lung Cancer
Maruja E. Lira∗, Tae Min Kim†, ‡, Donghui Huang∗, Shibing Deng∗, Youngil Koh‡, Bogun Jang§,Heounjeong Go§, Se-Hoon Lee†, ‡, Doo Hyun Chung§, Woo Ho Kim§, Eric F.P.M. Schoenmakers∗, Yoon-La Choi¶, Keunchil Park‖, Jin Seok Ahn‖, Jong-Mu Sun‖, Myung-Ju Ahn‖, Dong-Wan Kim†, ‡, , , ,Mao Mao∗
Anaplastic lymphoma kinase gene (ALK) fusions have been identified in approximately 5% of non-small-cell lung carcinomas (NSCLCs) and define a distinct subpopulation of patients with lung cancer who are highly responsive to ALK kinase inhibitors, such as crizotinib. Because of this profound therapeutic implication, the latest National Comprehensive Cancer Network Clinical Practice Guidelines in Oncology recommend upfront ALK screening for all patients with NSCLC. The Food and Drug Administration–approved companion diagnostic test (ie, fluorescence in situ hybridization) for identification of ALK-positive patients, however, is complex and has considerable limitations in terms of cost and throughput, making it difficult to screen many patients. To explore alternative screening modalities for detecting ALK fusions, we designed a combination of two transcript-based assays to detect for presence or absence of ALK fusions using NanoString’s nCounter technology. By using this combined gene expression and ALK fusion detection strategy, we developed a multiplexed assay with a quantitative scoring modality that is highly sensitive, reproducible, and capable of detecting low-abundant ALK fusion transcripts, even in samples with a low tumor cell content. In 66 archival NSCLC samples, our results were highly concordant to prior results obtained by fluorescence in situ hybridization and IHC. Our assay offers a cost-effective, easy-to-perform, high-throughput, and FFPE-compatible screening alternative for detection of ALK fusions.
