Discordance between Fluorescence In Situ Hybridization and Immunohistochemistry Analysis of Anaplastic Lymphoma Kinase Rearrangement in Indian Patients with Non-Small Cell Lung Cancer

Abstract

Aims This study aims to evaluate the incidence of anaplastic lymphoma kinase (ALK) mutation in nonsmall cell lung cancer (NSCLC) incorporating fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) methods and to look for any discordance.

Methods We evaluated 101 samples obtained from an enriched cohort of NSCLCs patients from the Army Hospital Research and Referral, New Delhi, India, between November 2016 and November 2018. IHC was performed using the highly-sensitive D5F3 rabbit monoclonal primary antibody. FISH was performed with dual-color, break-apart probe (ZytoLight SPEC) on formalin-fixed, and paraffin-embedded tissue. Discordance between IHC and FISH for ALK rearrangements was evaluated. Pearson correlation coefficient (r) was performed to identify any association of ALK presence (by IHC and FISH) with smoking brain metastasis, programmed death-ligand (PD-L1) expression, pleural effusion, and histopathological subtype.

Results A total of 7.92% (8/101) cases tested by IHC and 9.9% (10/101) cases tested by FISH were positive for ALK rearrangement. Of 93 ALK IHC-negative cases, 4 were ALK FISH-positive, whereas of 91 ALK FISH-negative cases, 4 were ALK IHC-positive cases. The correlation analysis demonstrated no or very weak correlation in ALK mutations by IHC or FISH with smoking, brain metastasis, PD-L1 expression, pleural effusion, and histopathological examination, except a weak positive correlation (r = 0.33) observed between brain metastasis and ALK rearrangement identified by FISH.

Conclusions Our study demonstrated a somewhat similar incidence of ALK FISH-positive cases and ALK IHC-positive cases, though the incidence was numerically higher for ALK-FISH method.

Keywords

anaplastic lymphoma kinase rearrangement - fluorescence in situ hybridization - immunohistochemistry - non-small cell lung cancer

Publikationsverlauf

Artikel online veröffentlicht:
14. Dezember 2020

• References

• 1 Scattone A, Catino A, Schirosi L. et al. Discordance between FISH, IHC, and NGS analysis of ALK status in advanced non-small cell lung cancer (NSCLC): a brief report of 7 cases. Transl Oncol 2019; 12 (02) 389-395
• 2 Moatter S, Anwar N, Moatter T, Pervez S. Frequency of ALK rearrangement by FISH testing and its correlation with ALK-IHC in adenocarcinoma of primary lung origin. Asian Pac J Cancer Prev 2018; 19 (06) 1735-1738
• 3 Cabillic F, Hofman P, Ilie M. et al. ALK IHC and FISH discordant results in patients with NSCLC and treatment response: for discussion of the question-to treat or not to treat?. ESMO Open 2018; 3 (06) e000419
• 4 Horn L, Pao W. EML4-ALK: honing in on a new target in non-small-cell lung cancer. J Clin Oncol 2009; 27 (26) 4232-4235
• 5 Minca EC, Portier BP, Wang Z. et al. ALK status testing in non-small cell lung carcinoma: correlation between ultrasensitive IHC and FISH. J Mol Diagn 2013; 15 (03) 341-346
• 6 Ross JS, Ali SM, Fasan O. et al. ALK fusions in a wide variety of tumor types respond to anti-ALK targeted therapy. Oncologist 2017; 22 (12) 1444-1450
• 7 Luk PP, Selinger CI, Mahar A, Cooper WA. Biomarkers for ALK and ROS 1 in lung cancer: Immunohistochemistry and fluorescent in situ hybridization. Arch Pathol Lab Med 2018; 142 (08) 922-928
• 8 Sullivan HC, Fisher KE, Hoffa AL. et al. The role of immunohistochemical analysis in the evaluation of EML4-ALK gene rearrangement in lung cancer. Appl Immunohistochem Mol Morphol 2015; 23 (04) 239-244
• 9 Uruga H, Mino-Kenudson M. ALK (D5F3) CDx: an immunohistochemistry assay to identify ALK-positive NSCLC patients. Pharm Genomics Pers Med 2018; 11: 147-155
• 10 Gainor JF, Varghese AM, Ou SH. et al. A retrospective analysis of the prevalence of EGFR or KRAS mutations in patients (pts) with crizotinib-naïve and crizotinib-resistant, ALK-positive non-small cell lung cancer (NSCLC). J Clin Oncol 2013; 31 (Suppl. 15) 8083
• 11 Gainor JF, Varghese AM, Ou SH. et al. ALK rearrangements are mutually exclusive with mutations in EGFR or KRAS: an analysis of 1,683 patients with non-small cell lung cancer. Clin Cancer Res 2013; 19 (15) 4273-4281
• 12 Yang JJ, Zhang XC, Su J. et al. Lung cancers with concomitant EGFR mutations and ALK rearrangements: diverse responses to EGFR-TKI and crizotinib in relation to diverse receptors phosphorylation. Clin Cancer Res 2014; 20 (05) 1383-1392
• 13 Yi ES, Boland JM, Maleszewski JJ. et al. Correlation of IHC and FISH for ALK gene rearrangement in non-small cell lung carcinoma: IHC score algorithm for FISH. J Thorac Oncol 2011; 6 (03) 459-465
• 14 Dearden S, Stevens J, Wu YL, Blowers D. Mutation incidence and coincidence in non small-cell lung cancer: meta-analyses by ethnicity and histology (mutMap). Ann Oncol 2013; 24 (09) 2371-2376
• 15 Doval D, Prabhash K, Patil S. et al. Clinical and epidemiological study of EGFR mutations and EML4-ALK fusion genes among Indian patients with adenocarcinoma of the lung. OncoTargets Ther 2015; 8: 117-123
• 16 Desai SS, Shah AS, Prabhash K, Jambhekar NA. A year of anaplastic large cell kinase testing for lung carcinoma: pathological and technical perspectives. Indian J Cancer 2013; 50 (02) 80-86
• 17 Rangachari D, Yamaguchi N, VanderLaan PA. et al. Brain metastases in patients with EGFR-mutated or ALK-rearranged non-small-cell lung cancers. Lung Cancer 2015; 88 (01) 108-111
• 18 Guérin A, Sasane M, Zhang J. et al. Brain metastases in patients with ALK+ non-small cell lung cancer: clinical symptoms, treatment patterns and economic burden. J Med Econ 2015; 18 (04) 312-322
• 19 Chatterjee K, Bhowmik R, Chattopadhyay B. Regional reporting of the incidence of anaplastic lymphoma kinase mutation in 379 non-small-cell lung cancer patients from Kolkata: using immunohistochemistry as the diagnostic modality in a significant subset. South Asian J Cancer 2017; 6 (04) 169-170
• 20 To KF, Tong JH, Yeung KS. et al. Detection of ALK rearrangement by immunohistochemistry in lung adenocarcinoma and the identification of a novel EML4-ALK variant. J Thorac Oncol 2013; 8 (07) 883-891
• 21 Hsiao SH, Chung CL, Chou YT, Lee HL, Lin SE, Liu HE. Identification of subgroup patients with stage IIIB/IV non-small cell lung cancer at higher risk for brain metastases. Lung Cancer 2013; 82 (02) 319-323
• 22 Griesinger F, Roeper J, Pöttgen C, Willborn KC, Eberhardt WEE. Brain metastases in ALK-positive NSCLC - time to adjust current treatment algorithms. Oncotarget 2018; 9 (80) 35181-35194
• 23 Lan B, Ma C, Zhang C. et al. Association between PD-L1 expression and driver gene status in non-small-cell lung cancer: a meta-analysis. Oncotarget 2018; 9 (07) 7684-7699
• 24 Furrukh M. Tobacco smoking and lung cancer: perception-changing facts. Sultan Qaboos Univ Med J 2013; 13 (03) 345-358
• 25 Carter J, Miller JA, Feller-Kopman D, Ettinger D. et al. Molecular Profiling of Malignant Pleural Effusion in Metastatic Non-Small-Cell Lung Carcinoma. The Effect of Preanalytical Factors. Ann Am Thorac Soc 2017; 14 (07) 1169-1176
• 26 Wang H, Zhang W, Wang K, Li X. Correlation between EML4-ALK, EGFR and clinicopathological features based on IASLC/ATS/ERS classification of lung adenocarcinoma. Medicine (Baltimore) 2018; 97 (26) e11116
• 27 da Cunha Santos G, Shepherd FA, Tsao MS. EGFR mutations and lung cancer. Annu Rev Pathol 2011; 6: 49-69
• 28 Chougule A, Prabhash K, Noronha V. et al. Frequency of EGFR mutations in 907 lung adenocarcinoma patients of Indian ethnicity. PLoS One 2013; 8 (10) e76164
• 29 Zhou J, Song XB, He H, Zhou Y, Lu XJ, Ying BW. Prevalence and clinical profile of EGFR mutation in non- small-cell lung carcinoma patients in Southwest China. Asian Pac J Cancer Prev 2016; 17 (03) 965-971