PURPOSE: EML4-ALK is a fusion-type protein tyrosine kinase that is generated by inv(2)(p21p23) in the genome of non-small cell lung cancer (NSCLC). To allow sensitive detection of EML4-ALK fusion transcripts, we have now developed a multiplex reverse transcription-PCR (RT-PCR) system that captures all in-frame fusions between the two genes. EXPERIMENTAL DESIGN: Primers were designed to detect all possible in-frame fusions of EML4 to exon 20 of ALK, and a single-tube multiplex RT-PCR assay was done with total RNA from 656 solid tumors of the lung (n = 364) and 10 other organs. RESULTS: From consecutive lung adenocarcinoma cases (n = 253), we identified 11 specimens (4.35%) positive for fusion transcripts, 9 of which were positive for the previously identified variants 1, 2, and 3. The remaining two specimens harbored novel transcript isoforms in which exon 14 (variant 4) or exon 2 (variant 5) of EML4 was connected to exon 20 of ALK. No fusion transcripts were detected for other types of lung cancer (n = 111) or for tumors from 10 other organs (n = 292). Genomic rearrangements responsible for the fusion events in NSCLC cells were confirmed by genomic PCR analysis and fluorescence in situ hybridization. The novel isoforms of EML4-ALK manifested marked oncogenic activity, and they yielded a pattern of cytoplasmic staining with fine granular foci in immunohistochemical analysis of NSCLC specimens. CONCLUSIONS: These data reinforce the importance of accurate diagnosis of EML4-ALK-positive tumors for the optimization of treatment strategies.
PURPOSE:EML4-ALK is a fusion-type protein tyrosine kinase that is generated by inv(2)(p21p23) in the genome of non-small cell lung cancer (NSCLC). To allow sensitive detection of EML4-ALK fusion transcripts, we have now developed a multiplex reverse transcription-PCR (RT-PCR) system that captures all in-frame fusions between the two genes. EXPERIMENTAL DESIGN: Primers were designed to detect all possible in-frame fusions of EML4 to exon 20 of ALK, and a single-tube multiplex RT-PCR assay was done with total RNA from 656 solid tumors of the lung (n = 364) and 10 other organs. RESULTS: From consecutive lung adenocarcinoma cases (n = 253), we identified 11 specimens (4.35%) positive for fusion transcripts, 9 of which were positive for the previously identified variants 1, 2, and 3. The remaining two specimens harbored novel transcript isoforms in which exon 14 (variant 4) or exon 2 (variant 5) of EML4 was connected to exon 20 of ALK. No fusion transcripts were detected for other types of lung cancer (n = 111) or for tumors from 10 other organs (n = 292). Genomic rearrangements responsible for the fusion events in NSCLC cells were confirmed by genomic PCR analysis and fluorescence in situ hybridization. The novel isoforms of EML4-ALK manifested marked oncogenic activity, and they yielded a pattern of cytoplasmic staining with fine granular foci in immunohistochemical analysis of NSCLC specimens. CONCLUSIONS: These data reinforce the importance of accurate diagnosis of EML4-ALK-positive tumors for the optimization of treatment strategies.
Authors: Alice T Shaw; Beow Y Yeap; Benjamin J Solomon; Gregory J Riely; Justin Gainor; Jeffrey A Engelman; Geoffrey I Shapiro; Daniel B Costa; Sai-Hong I Ou; Mohit Butaney; Ravi Salgia; Robert G Maki; Marileila Varella-Garcia; Robert C Doebele; Yung-Jue Bang; Kimary Kulig; Paulina Selaru; Yiyun Tang; Keith D Wilner; Eunice L Kwak; Jeffrey W Clark; A John Iafrate; D Ross Camidge Journal: Lancet Oncol Date: 2011-09-18 Impact factor: 41.316
Authors: Johannes M Heuckmann; Michael Hölzel; Martin L Sos; Stefanie Heynck; Hyatt Balke-Want; Mirjam Koker; Martin Peifer; Jonathan Weiss; Christine M Lovly; Christian Grütter; Daniel Rauh; William Pao; Roman K Thomas Journal: Clin Cancer Res Date: 2011-09-26 Impact factor: 12.531
Authors: Mari Mino-Kenudson; Lucian R Chirieac; Kenny Law; Jason L Hornick; Neal Lindeman; Eugene J Mark; David W Cohen; Bruce E Johnson; Pasi A Jänne; A John Iafrate; Scott J Rodig Journal: Clin Cancer Res Date: 2010-02-23 Impact factor: 12.531
Authors: Frank S Ong; Kingshuk Das; Jay Wang; Hana Vakil; Jane Z Kuo; Wendell-Lamar B Blackwell; Stephen W Lim; Mark O Goodarzi; Kenneth E Bernstein; Jerome I Rotter; Wayne W Grody Journal: Expert Rev Mol Diagn Date: 2012-07 Impact factor: 5.225