INTRODUCTION: A subset of lung cancers harbors an EML4-ALK (echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase) gene fusion, and detecting this subset may hold therapeutic implications. Many prior studies used fluorescence in situ hybridization (FISH) analysis for this detection, but FISH may have disadvantages including signal decay and dark-field examination that may obscure tissue architecture. In this study, we explored the potential of the ALK-break-apart chromogenic in situ hybridization (CISH) method to detect ALK-rearranged lung cancer. METHODS: We examined 15 lung adenocarcinomas with reverse-transcriptase polymerase chain reaction-proven EML4-ALK fusion transcripts and 30 ALK-negative cases. One hundred tumor cells were evaluated by CISH and FISH for each case, and a detailed signal profile was recorded and compared. RESULTS: CISH preserved tissue architecture and cytomorphology considerably and facilitated the signal evaluation using a routine light microscope. Positive rearrangement signals (splits or isolated 3' signals) were identified in 13 to 78% (mean ± SD, 41% ± 19%) of tumor cells in the ALK-positive cohort and in 0 to 15% (mean ± SD, 6% ± 4%) of cells in the ALK-negative cohort. The two groups were best separated by a cutoff value of 20%, with a sensitivity of 93% and a specificity of 100%. The only false-negative tumor having only 13% CISH-positive cells displayed predominantly (76%) isolated 5' signals unaccompanied by 3' signals. FISH showed largely similar signal profiles, and the results were completely concordant with CISH. CONCLUSIONS: We have successfully introduced CISH for diagnosing EML4-ALK-positive lung adenocarcinoma. This method allows simultaneous visualization of genetics and tumor cytomorphology and facilitates the molecular evaluation and could be applicable in clinical practice to detect lung cancer that may be responsive to ALK inhibitors.
INTRODUCTION: A subset of lung cancers harbors an EML4-ALK (echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase) gene fusion, and detecting this subset may hold therapeutic implications. Many prior studies used fluorescence in situ hybridization (FISH) analysis for this detection, but FISH may have disadvantages including signal decay and dark-field examination that may obscure tissue architecture. In this study, we explored the potential of the ALK-break-apart chromogenic in situ hybridization (CISH) method to detect ALK-rearranged lung cancer. METHODS: We examined 15 lung adenocarcinomas with reverse-transcriptase polymerase chain reaction-proven EML4-ALK fusion transcripts and 30 ALK-negative cases. One hundred tumor cells were evaluated by CISH and FISH for each case, and a detailed signal profile was recorded and compared. RESULTS:CISH preserved tissue architecture and cytomorphology considerably and facilitated the signal evaluation using a routine light microscope. Positive rearrangement signals (splits or isolated 3' signals) were identified in 13 to 78% (mean ± SD, 41% ± 19%) of tumor cells in the ALK-positive cohort and in 0 to 15% (mean ± SD, 6% ± 4%) of cells in the ALK-negative cohort. The two groups were best separated by a cutoff value of 20%, with a sensitivity of 93% and a specificity of 100%. The only false-negative tumor having only 13% CISH-positive cells displayed predominantly (76%) isolated 5' signals unaccompanied by 3' signals. FISH showed largely similar signal profiles, and the results were completely concordant with CISH. CONCLUSIONS: We have successfully introduced CISH for diagnosing EML4-ALK-positive lung adenocarcinoma. This method allows simultaneous visualization of genetics and tumor cytomorphology and facilitates the molecular evaluation and could be applicable in clinical practice to detect lung cancer that may be responsive to ALK inhibitors.
Authors: D Ross Camidge; Margaret Skokan; Porntip Kiatsimkul; Barbara Helfrich; Xian Lu; Anna E Barón; Nathan Schulte; DeLee Maxson; Dara L Aisner; Wilbur A Franklin; Robert C Doebele; Marileila Varella-Garcia Journal: Cancer Date: 2013-09-10 Impact factor: 6.860
Authors: Jin Sung Jang; Xiaoke Wang; Peter T Vedell; Ji Wen; Jinghui Zhang; David W Ellison; Jared M Evans; Sarah H Johnson; Ping Yang; William R Sukov; Andre M Oliveira; George Vasmatzis; Zhifu Sun; Jin Jen; Eunhee S Yi Journal: J Thorac Oncol Date: 2016-06-22 Impact factor: 15.609
Authors: Carlo Valentino; Samantha Kendrick; Nathalie Johnson; Randy Gascoyne; Wing C Chan; Dennis Weisenburger; Rita Braziel; James R Cook; Raymond Tubbs; Elias Campo; Andreas Rosenwald; German Ott; Jan Delabie; Elaine Jaffe; Wenjun Zhang; Patrick Brunhoeber; Hiro Nitta; Tom Grogan; Lisa Rimsza Journal: Am J Clin Pathol Date: 2013-02 Impact factor: 2.493
Authors: Shengxiang Ren; Fred R Hirsch; Marileila Varella-Garcia; Dara L Aisner; Theresa Boyle; Caicun Zhou; D Ross Camidge Journal: J Thorac Oncol Date: 2014-03 Impact factor: 15.609
Authors: Lien Tembuyser; Véronique Tack; Karen Zwaenepoel; Patrick Pauwels; Keith Miller; Lukas Bubendorf; Keith Kerr; Ed Schuuring; Erik Thunnissen; Elisabeth M C Dequeker Journal: PLoS One Date: 2014-11-11 Impact factor: 3.240
Authors: Pablo Martinez; Javier Hernández-Losa; Ma Ángeles Montero; Susana Cedrés; Josep Castellví; Alex Martinez-Marti; Natalia Tallada; Nuria Murtra-Garrell; Alejandro Navarro-Mendivill; Victor Rodriguez-Freixinos; Mercedes Canela; Santiago Ramon y Cajal; Enriqueta Felip Journal: PLoS One Date: 2013-01-24 Impact factor: 3.240