Rearrangements of the ROS1 gene occur in 1-2 % of non-small cell lung cancers (NSCLCs). Crizotinib, a highly effective inhibitor of ROS1 kinase activity, is now FDA-approved for the treatment of patients with advanced ROS1-positive NSCLC. Consequently, focus on ROS1 testing is growing. Most laboratories currently rely on fluorescence in situ hybridisation (FISH) assays using a dual-colour break-apart probe to detect ROS1 rearrangements. Given the rarity of these rearrangements in NSCLC, detection of elevated ROS1 protein levels by immunohistochemistry may provide cost-effective screening prior to confirmatory FISH testing. Non-in situ testing approaches also hold potential as stand-alone methods or complementary tests, including multiplex real-time PCR assays and next-generation sequencing (NGS) platforms which include commercial test kits covering a range of fusion genes. In order to ensure high-quality biomarker testing, appropriate tissue handling, adequate control materials and participation in external quality assessment programmes are essential, irrespective of the testing technique employed. ROS1 testing is often only considered after negative tests for EGFR mutation and ALK gene rearrangement, based on the assumption that these oncogenic driver events tend to be exclusive. However, as the use of ROS1 inhibitors becomes routine, accurate and timely detection of ROS1 gene rearrangements will be critical for the optimal treatment of patients with NSCLC. As NGS techniques are introduced into routine diagnostic practice, ROS1 fusion gene testing will be provided as part of the initial testing package.

Center of Predictive Molecular Medicine University Foundation Chieti Italy

Cerrahpasa Medical Faculty Istanbul University Istanbul Turkey

Department of Pathology Aberdeen University Medical School Aberdeen UK

Department of Pathology and Cytology Karolinska University Hospital Stockholm Sweden

Department of Pathology and Laboratory Medicine King Faisal Specialist Hospital and Research Centre Riyadh Saudi Arabia

Department of Pathology Centre Jean Perrin Clermont Ferrand France

Department of Pathology VU University Medical Centre Amsterdam The Netherlands

Institute of Pathology Charité Campus Mitte Berlin Germany

Institute of Pathology University Hospital Antwerp Edegem Belgium

Institute of Pathology University Hospital Basel Basel Switzerland

Institute of Pathology University Hospital Cologne and Network Genomic Medicine Cologne Germany

Laboratorio de Dianas Terapéuticas Hospital Universitario HM Sanchinarro C Oña 10 28050 Madrid Spain

The Fingerland Department of Pathology Charles University Faculty of Medicine and Faculty Hospital in Hradec Kralove Hradec Kralove Czech Republic

Unit of Pathologic Anatomy Azienda USL Valle d'Aosta Aosta Italy

Zobrazit více v PubMed

Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65(2):87–108. PubMed

Rosell R, Carcereny E, Gervais R, Vergnenegre A, Massuti B, Felip E, Palmero R, Garcia-Gomez R, Pallares C, Sanchez JM, Porta R, Cobo M, Garrido P, Longo F, Moran T, Insa A, De Marinis F, Corre R, Bover I, Illiano A, Dansin E, de Castro J, Milella M, Reguart N, Altavilla G, Jimenez U, Provencio M, Moreno MA, Terrasa J, Munoz-Langa J, Valdivia J, Isla D, Domine M, Molinier O, Mazieres J, Baize N, Garcia-Campelo R, Robinet G, Rodriguez-Abreu D, Lopez-Vivanco G, Gebbia V, Ferrera-Delgado L, Bombaron P, Bernabe R, Bearz A, Artal A, Cortesi E, Rolfo C, Sanchez-Ronco M, Drozdowskyj A, Queralt C, de Aguirre I, Ramirez JL, Sanchez JJ, Molina MA, Taron M, Paz-Ares L. Spanish Lung Cancer Group in collaboration with Groupe Francais de P-C, Associazione Italiana Oncologia T Erlotinib versus standard chemotherapy as first-line treatment for European patients with advanced EGFR mutation-positive non-small-cell lung cancer (EURTAC): a multicentre, open-label, randomised phase 3 trial. Lancet Oncol. 2012;13(3):239–246. PubMed

Solomon BJ, Mok T, Kim DW, Wu YL, Nakagawa K, Mekhail T, Felip E, Cappuzzo F, Paolini J, Usari T, Iyer S, Reisman A, Wilner KD, Tursi J, Blackhall F, Investigators P. First-line crizotinib versus chemotherapy in ALK-positive lung cancer. N Engl J Med. 2014;371(23):2167–2177. PubMed

Savas P, Hughes B, Solomon B. Targeted therapy in lung cancer: IPASS and beyond, keeping abreast of the explosion of targeted therapies for lung cancer. J Thorac Dis. 2013;5(Suppl 5):S579–S592. PubMed PMC

Shaw AT, Forcione DG, Digumarthy SR, Iafrate AJ. Case records of the Massachusetts General Hospital. Case 21-2011. A 31-year-old man with ALK-positive adenocarcinoma of the lung. N Engl J Med. 2011;365(2):158–167. PubMed

Kris MG, Johnson BE, Berry LD, Kwiatkowski DJ, Iafrate AJ, Wistuba II, Varella-Garcia M, Franklin WA, Aronson SL, Su P-F. Using multiplexed assays of oncogenic drivers in lung cancers to select targeted drugs. Jama. 2014;311(19):1998–2006. PubMed PMC

Network TCGAR (2014) Comprehensive molecular profiling of lung adenocarcinoma. Nature 511 (7511):543-550. doi: 10.1038/nature13385 http://www.nature.com/nature/journal/v511/n7511/abs/nature13385.html#supplementary-information PubMed PMC

Shaw AT, Ou SH, Bang YJ, Camidge DR, Solomon BJ, Salgia R, Riely GJ, Varella-Garcia M, Shapiro GI, Costa DB, Doebele RC, Le LP, Zheng Z, Tan W, Stephenson P, Shreeve SM, Tye LM, Christensen JG, Wilner KD, Clark JW, Iafrate AJ. Crizotinib in ROS1-rearranged non-small-cell lung cancer. N Engl J Med. 2014;371(21):1963–1971. PubMed PMC

Bergethon K, Shaw AT, Ou SH, Katayama R, Lovly CM, McDonald NT, Massion PP, Siwak-Tapp C, Gonzalez A, Fang R, Mark EJ, Batten JM, Chen H, Wilner KD, Kwak EL, Clark JW, Carbone DP, Ji H, Engelman JA, Mino-Kenudson M, Pao W, Iafrate AJ. ROS1 rearrangements define a unique molecular class of lung cancers. J Clin Oncol. 2012;30(8):863–870. PubMed PMC

Pfizer. (2016) Crizotinib Summary of Product Characteristics.

Pfizer. (2016) Crizotinib Prescribing Information.

FDA (2016) FDA expands use of Xalkori to treat rare form of advanced non-small cell lung cancer. http://www.fda.gov/newsevents/newsroom/pressannouncements/ucm490329.htm.

Shibuya M, Hanafusa H, Balduzzi PC. Cellular sequences related to three new onc genes of avian sarcoma virus (fps, yes, and ros) and their expression in normal and transformed cells. J Virol. 1982;42(1):143–152. PubMed PMC

Charest A, Lane K, McMahon K, Park J, Preisinger E, Conroy H, Housman D. Fusion of FIG to the receptor tyrosine kinase ROS in a glioblastoma with an interstitial del (6) (q21q21) Genes Chromosom Cancer. 2003;37(1):58–71. PubMed

Acquaviva J, Wong R, Charest A. The multifaceted roles of the receptor tyrosine kinase ROS in development and cancer. Biochim Biophys Acta. 2009;1795(1):37–52. PubMed

Davies KD, Doebele RC. Molecular pathways: ROS1 fusion proteins in cancer. Clin Cancer Res. 2013;19(15):4040–4045. PubMed PMC

Kim MH, Shim HS, Kang DR, Jung JY, Lee CY, Kim DJ, Lee JG, Bae MK, Kim HR, Lim SM, Kim EY, Park JS, Chung KY, Kim HJ, Kim JH, Cho BC. Clinical and prognostic implications of ALK and ROS1 rearrangements in never-smokers with surgically resected lung adenocarcinoma. Lung Cancer. 2014;83(3):389–395. PubMed

Govindan R, Ding L, Griffith M, Subramanian J, Dees ND, Kanchi KL, Maher CA, Fulton R, Fulton L, Wallis J, Chen K, Walker J, McDonald S, Bose R, Ornitz D, Xiong D, You M, Dooling DJ, Watson M, Mardis ER, Wilson RK. Genomic landscape of non-small cell lung cancer in smokers and never-smokers. Cell. 2012;150(6):1121–1134. PubMed PMC

Rikova K, Guo A, Zeng Q, Possemato A, Yu J, Haack H, Nardone J, Lee K, Reeves C, Li Y, Hu Y, Tan Z, Stokes M, Sullivan L, Mitchell J, Wetzel R, Macneill J, Ren JM, Yuan J, Bakalarski CE, Villen J, Kornhauser JM, Smith B, Li D, Zhou X, Gygi SP, Gu TL, Polakiewicz RD, Rush J, Comb MJ. Global survey of phosphotyrosine signaling identifies oncogenic kinases in lung cancer. Cell. 2007;131(6):1190–1203. PubMed

Rimkunas VM, Crosby KE, Li D, Hu Y, Kelly ME, Gu TL, Mack JS, Silver MR, Zhou X, Haack H. Analysis of receptor tyrosine kinase ROS1-positive tumors in non-small cell lung cancer: identification of a FIG-ROS1 fusion. Clin Cancer Res. 2012;18(16):4449–4457. PubMed

Seo JS, Ju YS, Lee WC, Shin JY, Lee JK, Bleazard T, Lee J, Jung YJ, Kim JO, Shin JY, Yu SB, Kim J, Lee ER, Kang CH, Park IK, Rhee H, Lee SH, Kim JI, Kang JH, Kim YT. The transcriptional landscape and mutational profile of lung adenocarcinoma. Genome Res. 2012;22(11):2109–2119. PubMed PMC

Takeuchi K, Soda M, Togashi Y, Suzuki R, Sakata S, Hatano S, Asaka R, Hamanaka W, Ninomiya H, Uehara H, Lim Choi Y, Satoh Y, Okumura S, Nakagawa K, Mano H, Ishikawa Y. RET, ROS1 and ALK fusions in lung cancer. Nat Med. 2012;18(3):378–381. PubMed

Yoshida A, Kohno T, Tsuta K, Wakai S, Arai Y, Shimada Y, Asamura H, Furuta K, Shibata T, Tsuda H. ROS1-rearranged lung cancer: a clinicopathologic and molecular study of 15 surgical cases. Am J Surg Pathol. 2013;37(4):554–562. PubMed

Birchmeier C, Sharma S, Wigler M. Expression and rearrangement of the ROS1 gene in human glioblastoma cells. Proc Natl Acad Sci U S A. 1987;84(24):9270–9274. PubMed PMC

Jun HJ, Johnson H, Bronson RT, de Feraudy S, White F, Charest A. The oncogenic lung cancer fusion kinase CD74-ROS activates a novel invasiveness pathway through E-Syt1 phosphorylation. Cancer Res. 2012;72(15):3764–3774. PubMed PMC

Charest A, Kheifets V, Park J, Lane K, McMahon K, Nutt CL, Housman D. Oncogenic targeting of an activated tyrosine kinase to the Golgi apparatus in a glioblastoma. Proc Natl Acad Sci U S A. 2003;100(3):916–921. PubMed PMC

Mazieres J, Zalcman G, Crino L, Biondani P, Barlesi F, Filleron T, Dingemans AM, Lena H, Monnet I, Rothschild SI, Cappuzzo F, Besse B, Thiberville L, Rouviere D, Dziadziuszko R, Smit EF, Wolf J, Spirig C, Pecuchet N, Leenders F, Heuckmann JM, Diebold J, Milia JD, Thomas RK, Gautschi O. Crizotinib therapy for advanced lung adenocarcinoma and a ROS1 rearrangement: results from the EUROS1 cohort. J Clin Oncol. 2015;33(9):992–999. PubMed

Moro-Sibilot D, Faivre L, Zalcman G, Perol M, Mazières J, Barlesi F, Otto J, Monnet I, Cortot A, Wislez M, Léna H, Souquet PJ, Lantuejoul S, Rouquette I, Mcleer-Florin A, Ferretti G, Hoog-Labouret N, Nowak F, Jimenez M, Vassal G. Crizotinib in patients with ROS1 NSCLC. Preliminary results of the AcSé trial. J Thorac Oncol. 2015;10(9):S261–S406.

Goto K, Yang JCH, Kim D-W, Lu S, Seto T, Yang J-J, Yamamoto N, Ahn M-J, Takahashi T, Yamanaka T, Kemner A, Roychowdhury D, Paolini J, Wilner K, Wu Y-L. Phase II study of crizotinib in East Asian patients (pts) with ROS1-positive advanced non-small cell lung cancer (NSCLC) J Clin Oncol. 2016;34(15 suppl.):Abstr. 9022. PubMed

Shaw AT, Kim DW, Nakagawa K, Seto T, Crino L, Ahn MJ, De Pas T, Besse B, Solomon BJ, Blackhall F, Wu YL, Thomas M, O'Byrne KJ, Moro-Sibilot D, Camidge DR, Mok T, Hirsh V, Riely GJ, Iyer S, Tassell V, Polli A, Wilner KD, Janne PA. Crizotinib versus chemotherapy in advanced ALK-positive lung cancer. N Engl J Med. 2013;368(25):2385–2394. PubMed

Dietel M, Bubendorf L, Dingemans AM, Dooms C, Elmberger G, Garcia RC, Kerr KM, Lim E, Lopez-Rios F, Thunnissen E, Van Schil PE, von Laffert M. Diagnostic procedures for non-small-cell lung cancer (NSCLC): recommendations of the European Expert Group. Thorax. 2016;71(2):177–184. PubMed PMC

Thunnissen E, Bubendorf L, Dietel M, Elmberger G, Kerr K, Lopez-Rios F, Moch H, Olszewski W, Pauwels P, Penault-Llorca F, Rossi G. EML4-ALK testing in non-small cell carcinomas of the lung: a review with recommendations. Virchows Arch. 2012;461(3):245–257. PubMed PMC

Kerr KM, Bubendorf L, Edelman MJ, Marchetti A, Mok T, Novello S, O'Byrne K, Stahel R, Peters S, Felip E, Panel M, Panel M. Second ESMO consensus conference on lung cancer: pathology and molecular biomarkers for non-small-cell lung cancer. Ann Oncol. 2014;25(9):1681–1690. PubMed

Thunnissen E, van der Oord K, den Bakker M. Prognostic and predictive biomarkers in lung cancer. A review. Virchows Arch. 2014;464(3):347–358. PubMed

Gainor JF, Shaw AT. Novel targets in non-small cell lung cancer: ROS1 and RET fusions. Oncologist. 2013;18(7):865–875. PubMed PMC

Mescam-Mancini L, Lantuejoul S, Moro-Sibilot D, Rouquette I, Souquet PJ, Audigier-Valette C, Sabourin JC, Decroisette C, Sakhri L, Brambilla E, McLeer-Florin A. On the relevance of a testing algorithm for the detection of ROS1-rearranged lung adenocarcinomas. Lung Cancer. 2014;83(2):168–173. PubMed

Shan L, Lian F, Guo L, Qiu T, Ling Y, Ying J, Lin D. Detection of ROS1 gene rearrangement in lung adenocarcinoma: comparison of IHC, FISH and real-time RT-PCR. PLoS One. 2015;10(3):e0120422. PubMed PMC

Jin Y, Sun PL, Kim H, Park E, Shim HS, Jheon S, Kim K, Lee CT, Chung JH. Erratum to: ROS1 gene rearrangement and copy number gain in non-small cell lung cancer. Virchows Arch. 2015;467(1):123. PubMed

Bozzetti C, Nizzoli R, Tiseo M, Squadrilli A, Lagrasta C, Buti S, Gasparro D, Zanoni D, Majori M, De Filippo M, Mazzoni F, Maddau C, Naldi N, Sammarelli G, Frati C, Pinto C, Ardizzoni A. ALK and ROS1 rearrangements tested by fluorescence in situ hybridization in cytological smears from advanced non-small cell lung cancer patients. Diagn Cytopathol. 2015;43(11):941–946. PubMed

Savic S, Bubendorf L. Role of fluorescence in situ hybridization in lung cancer cytology. Acta Cytol. 2012;56(6):611–621. PubMed

Boyle TA, Masago K, Ellison KE, Yatabe Y, Hirsch FR. ROS1 immunohistochemistry among major genotypes of non-small-cell lung cancer. Clin Lung Cancer. 2015;16(2):106–111. PubMed PMC

Cao B, Wei P, Liu Z, Bi R, Lu Y, Zhang L, Zhang J, Yang Y, Shen C, Du X, Zhou X. Detection of lung adenocarcinoma with ROS1 rearrangement by IHC, FISH, and RT-PCR and analysis of its clinicopathologic features. Onco Targets Ther. 2016;9:131–138. PubMed PMC

Sholl LM, Sun H, Butaney M, Zhang C, Lee C, Janne PA, Rodig SJ. ROS1 immunohistochemistry for detection of ROS1-rearranged lung adenocarcinomas. Am J Surg Pathol. 2013;37(9):1441–1449. PubMed PMC

Yoshida A, Tsuta K, Wakai S, Arai Y, Asamura H, Shibata T, Furuta K, Kohno T, Kushima R. Immunohistochemical detection of ROS1 is useful for identifying ROS1 rearrangements in lung cancers. Mod Pathol. 2014;27(5):711–720. PubMed

Rogers TM, Russell PA, Wright G, Wainer Z, Pang JM, Henricksen LA, Singh S, Stanislaw S, Grille J, Roberts E, Solomon B, Fox SB. Comparison of methods in the detection of ALK and ROS1 rearrangements in lung cancer. J Thorac Oncol. 2015;10(4):611–618. PubMed

Bruce B, Khanna G, Ren L, Landberg G, Jirstrom K, Powell C, Borczuk A, Keller ET, Wojno KJ, Meltzer P, Baird K, McClatchey A, Bretscher A, Hewitt SM, Khanna C. Expression of the cytoskeleton linker protein ezrin in human cancers. Clin Exp Metastasis. 2007;24(2):69–78. PubMed

Murakami Y, Mitsudomi T, Yatabe Y. A Screening Method for the ALK Fusion Gene in NSCLC. Front Oncol. 2012;2:24. PubMed PMC

Fischer AH, Schwartz MR, Moriarty AT, Wilbur DC, Souers R, Fatheree L, Booth CN, Clayton AC, Kurtyz DF, Padmanabhan V, Crothers BA. Immunohistochemistry practices of cytopathology laboratories: a survey of participants in the College of American Pathologists Nongynecologic Cytopathology Education Program. Arch Pathol Lab Med. 2014;138(9):1167–1172. PubMed

Kirbis IS, Maxwell P, Flezar MS, Miller K, Ibrahim M. External quality control for immunocytochemistry on cytology samples: a review of UK NEQAS ICC (cytology module) results. Cytopathology. 2011;22(4):230–237. PubMed

Lee SE, Lee B, Hong M, Song JY, Jung K, Lira ME, Mao M, Han J, Kim J, Choi YL. Comprehensive analysis of RET and ROS1 rearrangement in lung adenocarcinoma. Mod Pathol. 2015;28(4):468–479. PubMed

Zheng Z, Liebers M, Zhelyazkova B, Cao Y, Panditi D, Lynch KD, Chen J, Robinson HE, Shim HS, Chmielecki J, Pao W, Engelman JA, Iafrate AJ, Le LP. Anchored multiplex PCR for targeted next-generation sequencing. Nat Med. 2014;20(12):1479–1484. PubMed

Ribeiro-Silva A, Zhang H, Jeffrey SS. RNA extraction from ten year old formalin-fixed paraffin-embedded breast cancer samples: a comparison of column purification and magnetic bead-based technologies. BMC Mol Biol. 2007;8:118. PubMed PMC

Gruber K, Horn H, Kalla J, Fritz P, Rosenwald A, Kohlhaufl M, Friedel G, Schwab M, Ott G, Kalla C. Detection of rearrangements and transcriptional up-regulation of ALK in FFPE lung cancer specimens using a novel, sensitive, quantitative reverse transcription polymerase chain reaction assay. J Thorac Oncol. 2014;9(3):307–315. PubMed

Moskalev EA, Frohnauer J, Merkelbach-Bruse S, Schildhaus HU, Dimmler A, Schubert T, Boltze C, Konig H, Fuchs F, Sirbu H, Rieker RJ, Agaimy A, Hartmann A, Haller F. Sensitive and specific detection of EML4-ALK rearrangements in non-small cell lung cancer (NSCLC) specimens by multiplex amplicon RNA massive parallel sequencing. Lung Cancer. 2014;84(3):215–221. PubMed

Lira ME, Choi YL, Lim SM, Deng S, Huang D, Ozeck M, Han J, Jeong JY, Shim HS, Cho BC, Kim J, Ahn MJ, Mao M. A single-tube multiplexed assay for detecting ALK, ROS1, and RET fusions in lung cancer. J Mol Diagn. 2014;16(2):229–243. PubMed

Thermo Fisher Scientific. Oncomine™ Solid Tumour Fusion Transcript Kit Product Information. Available at https://www.thermofisher.com/uk/en/home/clinical/molecular-diagnostics/oncology-diagnostics.html

Swanton C, Govindan R. Clinical implications of genomic discoveries in lung cancer. N Engl J Med. 2016;374(19):1864–1873. PubMed

Cha YJ, Lee JS, Kim HR, Lim SM, Cho BC, Lee CY, Shim HS. Screening of ROS1 rearrangements in lung adenocarcinoma by immunohistochemistry and comparison with ALK rearrangements. PLoS One. 2014;9(7):e103333. PubMed PMC

Lindeman NI, Cagle PT, Beasley MB, Chitale DA, Dacic S, Giaccone G, Jenkins RB, Kwiatkowski DJ, Saldivar JS, Squire J, Thunnissen E, Ladanyi M, College of American Pathologists International Association for the Study of Lung C. Association for Molecular P Molecular testing guideline for selection of lung cancer patients for EGFR and ALK tyrosine kinase inhibitors: guideline from the College of American Pathologists, International Association for the Study of Lung Cancer, and Association for Molecular Pathology. J Mol Diagn. 2013;15(4):415–453. PubMed

Felip E, Concha Á, de Castro J, Gómez-Román J, Garrido P, Ramírez J, Isla D, Sanz J, Paz-Ares L, López-Ríos F. Biomarker testing in advanced non-small-cell lung cancer: a National Consensus of the Spanish Society of Pathology and the Spanish Society of Medical Oncology. Clin Transl Oncol. 2015;17(2):103–112. PubMed

Zhang N, Yang JJ, Zhang XC, Xie Z, Wang BC, Tu HY, Jiang BY, Wu YL. Responses to crizotinib in a patient with c-ros oncogene 1, receptor tyrosine kinase-positive advanced lung adenocarcinoma: A case report. Oncol Lett. 2014;8(6):2624–2626. PubMed PMC

Jurmeister P, Lenze D, Berg E, Mende S, Schaper F, Kellner U, Herbst H, Sers C, Budczies J, Dietel M, Hummel M, von Laffert M. Parallel screening for ALK, MET and ROS1 alterations in non-small cell lung cancer with implications for daily routine testing. Lung Cancer. 2015;87(2):122–129. PubMed

Bussolati G, Annaratone L, Medico E, D'Armento G, Sapino A. Formalin fixation at low temperature better preserves nucleic acid integrity. PLoS One. 2011;6(6):e21043. PubMed PMC

Bussolati G, Annaratone L, Maletta F. The pre-analytical phase in surgical pathology. Recent Results Cancer Res. 2015;199:1–13. PubMed

Tembuyser L, Tack V, Zwaenepoel K, Pauwels P, Miller K, Bubendorf L, Kerr K, Schuuring E, Thunnissen E, Dequeker EM. The relevance of external quality assessment for molecular testing for ALK positive non-small cell lung cancer: results from two pilot rounds show room for optimization. PLoS One. 2014;9(11):e112159. PubMed PMC

Arai Y, Totoki Y, Takahashi H, Nakamura H, Hama N, Kohno T, Tsuta K, Yoshida A, Asamura H, Mutoh M, Hosoda F, Tsuda H, Shibata T. Mouse model for ROS1-rearranged lung cancer. PLoS One. 2013;8(2):e56010. PubMed PMC

Cai W, Li X, Su C, Fan L, Zheng L, Fei K, Zhou C, Manegold C, Schmid-Bindert G. ROS1 fusions in Chinese patients with non-small-cell lung cancer. Ann Oncol. 2013;24(7):1822–1827. PubMed

Cheng H, Ye L, Xue L. Detection of ROS1 gene rearrangement by FISH and analysis of its clinical features in non-small cell lung cancer patients. Zhonghua Zhong Liu Za Zhi. 2014;36(10):751–754. PubMed

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. Identifying and targeting ROS1 gene fusions in non-small cell lung cancer. Clin Cancer Res. 2012;18(17):4570–4579. PubMed PMC

Fu S, Liang Y, Lin YB, Wang F, Huang MY, Zhang ZC, Wang J, Cen WJ, Shao JY. The Frequency and Clinical Implication of ROS1 and RET Rearrangements in Resected Stage IIIA-N2 Non-Small Cell Lung Cancer Patients. PLoS One. 2015;10(4):e0124354. PubMed PMC

Go H, Kim DW, Kim D, Keam B, Kim TM, Lee SH, Heo DS, Bang YJ, Chung DH. Clinicopathologic analysis of ROS1-rearranged non-small-cell lung cancer and proposal of a diagnostic algorithm. J Thorac Oncol. 2013;8(11):1445–1450. PubMed

Jin Y, Sun PL, Kim H, Park E, Shim HS, Jheon S, Kim K, Lee CT, Chung JH. ROS1 gene rearrangement and copy number gain in non-small cell lung cancer. Virchows Arch. 2015;466(1):45–52. PubMed

Karlsson A, Brunnstrom H, Lindquist KE, Jirstrom K, Jonsson M, Rosengren F, Reutersward C, Cirenajwis H, Borg A, Jonsson P, Planck M, Jonsson G, Staaf J. Mutational and gene fusion analyses of primary large cell and large cell neuroendocrine lung cancer. Oncotarget. 2015;6(26):22028–22037. PubMed PMC

Kim HR, Lim SM, Kim HJ, Hwang SK, Park JK, Shin E, Bae MK, Ou SH, Wang J, Jewell SS, Kang DR, Soo RA, Haack H, Kim JH, Shim HS, Cho BC. The frequency and impact of ROS1 rearrangement on clinical outcomes in never smokers with lung adenocarcinoma. Ann Oncol. 2013;24(9):2364–2370. PubMed

Kirita K, Izumo T, Matsumoto Y, Hiraishi Y, Tsuchida T. Bronchoscopic Re-biopsy for Mutational Analysis of Non-small Cell Lung Cancer. Lung. 2016 PubMed

Li C, Fang R, Sun Y, Han X, Li F, Gao B, Iafrate AJ, Liu XY, Pao W, Chen H, Ji H. Spectrum of oncogenic driver mutations in lung adenocarcinomas from East Asian never smokers. PLoS One. 2011;6(11):e28204. PubMed PMC

Matsuura S, Shinmura K, Kamo T, Igarashi H, Maruyama K, Tajima M, Ogawa H, Tanahashi M, Niwa H, Funai K, Kohno T, Suda T, Sugimura H. CD74-ROS1 fusion transcripts in resected non-small cell lung carcinoma. Oncol Rep. 2013;30(4):1675–1680. PubMed

Okamoto I, Sakai K, Morita S, Yoshioka H, Kaneda H, Takeda K, Hirashima T, Kogure Y, Kimura T, Takahashi T, Atagi S, Seto T, Sawa T, Yamamoto M, Satouchi M, Okuno M, Nagase S, Takayama K, Tomii K, Maeda T, Oizumi S, Fujii S, Akashi Y, Nishino K, Ebi N, Nakagawa K, Nakanishi Y, Nishio K. Multiplex genomic profiling of non-small cell lung cancers from the LETS phase III trial of first-line S-1/carboplatin versus paclitaxel/carboplatin: results of a West Japan Oncology Group study. Oncotarget. 2014;5(8):2293–2304. PubMed PMC

Scheffler M, Schultheis A, Teixido C, Michels S, Morales-Espinosa D, Viteri S, Hartmann W, Merkelbach-Bruse S, Fischer R, Schildhaus HU, Fassunke J, Sebastian M, Serke M, Kaminsky B, Randerath W, Gerigk U, Ko YD, Kruger S, Schnell R, Rothe A, Kropf-Sanchen C, Heukamp L, Rosell R, Buttner R, Wolf J. ROS1 rearrangements in lung adenocarcinoma: prognostic impact, therapeutic options and genetic variability. Oncotarget. 2015;6(12):10577–10585. PubMed PMC

Suehara Y, Arcila M, Wang L, Hasanovic A, Ang D, Ito T, Kimura Y, Drilon A, Guha U, Rusch V, Kris MG, Zakowski MF, Rizvi N, Khanin R, Ladanyi M. Identification of KIF5B-RET and GOPC-ROS1 fusions in lung adenocarcinomas through a comprehensive mRNA-based screen for tyrosine kinase fusions. Clin Cancer Res. 2012;18(24):6599–6608. PubMed PMC

Wang R, Zhang Y, Pan Y, Li Y, Hu H, Cai D, Li H, Ye T, Luo X, Zhang Y, Li B, Shen L, Sun Y, Chen H. Comprehensive investigation of oncogenic driver mutations in Chinese non-small cell lung cancer patients. Oncotarget. 2015;6(33):34300–34308. PubMed PMC

Warth A, Muley T, Dienemann H, Goeppert B, Stenzinger A, Schnabel PA, Schirmacher P, Penzel R, Weichert W. ROS1 expression and translocations in non-small-cell lung cancer: clinicopathological analysis of 1478 cases. Histopathology. 2014;65(2):187–194. PubMed

Zhang Q, Sun T, Kang P, Qian K, Deng B, Zhou J, Wang R, Jiang B, Li K, Liu F, Wu S, Tan Q. Combined analysis of rearrangement of ALK, ROS1, somatic mutation of EGFR, KRAS, BRAF, PIK3CA, and mRNA expression of ERCC1, TYMS, RRM1, TUBB3, EGFR in patients with non-small cell lung cancer and their clinical significance. Cancer Chemother Pharmacol. 2016;77(3):583–593. PubMed

Zhao C, Li X, Li J, Zhang Y, Ren S, Chen X, Zhou C. Detecting ALK, ROS1 and RET Fusion Genes in Cell Block Samples. Transl Oncol. 2014;7(3):363–367. PubMed PMC

Zhong S, Zhang H, Bai D, Gao D, Zheng J, Ding Y. Detection of ALK, ROS1 and RET fusion genes in non-small cell lung cancer patients and its clinicopathologic correlation. Zhonghua Bing Li Xue Za Zhi. 2015;44(9):639–643. PubMed

Drilon A, Wang L, Arcila ME, Balasubramanian S, Greenbowe JR, Ross JS, Stephens P, Lipson D, Miller VA, Kris MG, Ladanyi M, Rizvi NA. Broad, Hybrid Capture-Based Next-Generation Sequencing Identifies Actionable Genomic Alterations in Lung Adenocarcinomas Otherwise Negative for Such Alterations by Other Genomic Testing Approaches. Clin Cancer Res. 2015;21(16):3631–3639. PubMed PMC

Leary RJ, Kinde I, Diehl F, Schmidt K, Clouser C, Duncan C, Antipova A, Lee C, McKernan K, De La Vega FM, Kinzler KW, Vogelstein B, Diaz LA, Jr, Velculescu VE. Development of personalized tumor biomarkers using massively parallel sequencing. Sci Transl Med. 2010;2(20):20ra14. PubMed PMC

The leading role of pathology in assessing the somatic molecular alterations of cancer: Position Paper of the European Society of Pathology

A stitch in time saves nine: external quality assessment rounds demonstrate improved quality of biomarker analysis in lung cancer