Discordance between FISH, IHC, and NGS Analysis of ALK Status in Advanced Non–Small Cell Lung Cancer (NSCLC): a Brief Report of 7 Cases

Scattone, Anna; Catino, Annamaria; Schirosi, Laura; Caldarola, Lucia; Tommasi, Stefania; Lacalamita, Rosanna; Montagna, Elisabetta Sara; Galetta, Domenico; Serio, Gabriella; Zito, Francesco Alfredo; Mangia, Anita

doi:10.1016/j.tranon.2018.11.006

Cited by 26 publications

(24 citation statements)

References 44 publications

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“…importance of careful interpretation of the ALK FISH assay. Diverse diagnostic modalities are highly recommended in ambiguous cases to insure the proper treatment for patients eligible for TKI therapy [19][20][21][22][23]. Based on our findings we strongly recommend the inclusion of the highly challenging atypical pattern of 3′ signal attenuation into the official guidelines.…”

Section: Resultsmentioning

confidence: 74%

Attenuated isolated 3’ signal: A highly challenging therapy relevant ALK FISH pattern in NSCLC

et al. 2020

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Targeted therapies in the management of patients with lung cancer provide significantly better outcome compared to chemotherapy. Detection of the anaplastic lymphoma kinase (ALK) gene rearrangement has great predictive value for treatment with small molecule tyrosine kinase inhibitor (crizotinib and alectinib commonly). Fluorescent in situ hybridisation (FISH) assay is a basic diagnostic test designed for detecting ALK gene rearrangements. Although being considered as gold standard method by IASLC's guideline, it is often regarded as difficult and error prone. Our aim was to examine a unique atypical ALK FISH pattern, revealed during a systematic large-scale monitoring, which carries the great risk of misinterpretation, hence may result in loss of patients eligible for targeted therapy. Materials and Methods: Tissue and cytology samples from nearly one thousand patients with advanced stage nonsmall cell lung cancer (NSCLC, n = 996) were routinely examined by ALK FISH and immunohistochemistry (Ventana ALK-D5F3-CDx assay). Anchored Multiplex PCR based Next Generation Sequencing (AMP-NGS) was used to detect fusion gene transcripts in ambiguous cases. Results: Fifty-nine (5,9%) of the cases were positive with ALK FISH test. Three cases showed atypical pattern with a significantly reduced sized red (3′) signal and complete loss of green signals. Digital signal measurement confirmed this finding, showing consistent attenuation of 3′ signals throughout the tumours. In all three cases AMP-NGS and ALK IHC verified the presence of a fusion gene and expressed oncoprotein, respectively. Conclusion: Approximately 5% of the 59 ALK positive cases exhibited atypical attenuated isolated 3′ signal pattern. The immunohistochemistry and AMP-NGS examinations helped to clarify the presence of oncoprotein and the fusion gene, respectively. Our results emphasize the importance of extensive exploration of the genetic background of any unexpected FISH finding to avoid false diagnosis. This enables clinicians to indicate the adequate therapy with higher efficiency for patients suffering from NSCLC.

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Section: Resultsmentioning

confidence: 74%

Attenuated isolated 3’ signal: A highly challenging therapy relevant ALK FISH pattern in NSCLC

et al. 2020

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“…23,26,[28][29][30][31][32] However, only a few studies investigated the reliability of NGS as compared with other traditional detection methods. [18][19][20][21][22][23] In this study, we compared NGS -a method that has become routinely used in clinical oncology due to its multiplex-ability-with IHC -an established method that is routinely used for the molecular diagnosis of ALK in the clinical setting. Based on our data, ALK detection using NGS and IHC was 77.3% to 78.9% concordant, which is slightly lower than a previous report that demonstrated an 87.3% concordance rate.…”

Section: Discussionmentioning

confidence: 99%

“…[16][17][18] In addition to mutation status, the details of ALK fusion gene partners can also be revealed by NGS. Although previous reports have explored the utility of NGS in detecting ALK rearrangements and indicated that NGS-based ALKpositive status may predict clinical benefit with crizotinib, [19][20][21][22][23] the association between ALK status assessed by NGS and therapeutic response from crizotinib has not been well validated. Studies with larger sample size are needed to establish the role of NGS in selecting patients eligible for crizotinib treatment.…”

Section: Introductionmentioning

confidence: 99%

<p>Comparison of Next-Generation Sequencing and Ventana Immunohistochemistry in Detecting ALK Rearrangements and Predicting the Efficacy of First-Line Crizotinib in Patients with Advanced Non-Small Cell Lung Cancer</p>

Zeng¹,

Li²,

Xu³

et al. 2020

OTT

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Introduction: Reliable diagnostic approaches to detect ALK rearrangement are critical for selecting patients eligible for crizotinib therapy. This study aimed to compare nextgeneration sequencing (NGS) and Ventana immunohistochemistry (IHC) in evaluating ALK rearrangements and evaluate their impact on first-line crizotinib efficacy. Patients and Methods: A total of 472 NSCLC patients were identified as ALK-positive by NGS and/or IHC between March 2014 and February 2020. The concordance of ALK detection, overall response rate (ORR), and progression-free survival (PFS) were analyzed for 319 patients who received front-line crizotinib. Results: First-line crizotinib (n=319) significantly prolonged PFS in comparison with chemotherapy (n=46; 12.0 vs 6.8 months; p<0.0001). Of the 76 crizotinib-treated patients whose ALK status was assessed by both NGS and IHC, 78.9% of the patients had concordant ALK status (NGS-positive/IHC-positive), 18.4% patients were NGS-positive but IHC-negative, and 2 patients were IHC-positive but NGS-negative. Different detection assays confer no statistical difference in ORR and PFS with first-line crizotinib. The ORR in NGS only, IHC only, and both NGS and IHC was 84.3%, 90.1%, and 88.1%, respectively, while PFS was 11.4, 13.0, and 11.0 months, respectively. The ORR in NGS-positive/IHC-positive and NGS-positive/IHC-negative patients was 85.4% and 92.8%, respectively. Compared to NGS-positive/IHC-positive patients, those with NGS-positive/IHC-negative results had a trend of shorter PFS but statistical significance was not reached (mPFS, 5.9 months vs 11.5 months, p=0.43). Conclusion: Our results demonstrate that ALK status detected by NGS and/or IHC is reliable in identifying patients with ALK-positive NSCLC who will benefit from ALK inhibitor therapy.

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“…NGS is technology with the ability to perform and capture data from millions of sequencing reactions simultaneously. All NGS platforms are able to catch the individual sequence of hundreds of millions of molecules at the genome or transcriptome level 28,[64][65][66][67] . One obstacle to initiating NGS in routine practice is the high initial costs as it method is currently more expensive than the methods above.…”

Section: Gene Rearrangement Detection In Nsclc Patientsmentioning

confidence: 99%

“…1) (ref. 26,67 ). IHC is still the most used and remains the gold standard in ALK rearrangement detection.…”

Section: Gene Rearrangement Detection In Nsclc Patientsmentioning

confidence: 99%

Gene rearrangement detection by next-generation sequencing in patients with non-small cell lung carcinoma

Brisudova¹,

Škarda²

2020

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub

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Non-small cell lung carcinoma (NSCLC) is the leading cause of cancer-related deaths worldwide. Various molecular markers in NSCLC patients have been developed, including gene rearrangements, currently used in therapeutic strategies. With increasing number of these molecular biomarkers of NSCLC, there is a demand for highly efficient methods for detecting mutations and translocations in treatable targets. Those currently available U.S. Food and Drug Administration (FDA) approved approaches, for example imunohistochemisty (IHC) and fluorescence in situ hybridization (FISH), are inadequate, due to sufficient quantity of material and long time duration. Next-generation massive parallel sequencing (NGS), with the ability to perform and capture data from millions of sequencing reactions simultaneously could resolve the problem. Thanks to gradual NGS introduction into clinical laboratories, screening time should be considerably shorter, which is very important for patients with advanced NSCLC. Moreover, only a minimum sample input is needed for achieving adequate results. NGS was compared to the current detection methods of ALK, ROS1, c-RET and c-MET rearrangements in NSCLC and a significant match, between IHC, FISH and NGS results, was found. Recent available researches have been carried out on a small numbers of patients. Verifying these results on larger patients cohort is important. This review sumarizes the literature on this subject and compares current possibilities of predictive gene rearrangements detection in patients with NSCLC.

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Discordance between FISH, IHC, and NGS Analysis of ALK Status in Advanced Non–Small Cell Lung Cancer (NSCLC): a Brief Report of 7 Cases

Cited by 26 publications

References 44 publications

Attenuated isolated 3’ signal: A highly challenging therapy relevant ALK FISH pattern in NSCLC

Attenuated isolated 3’ signal: A highly challenging therapy relevant ALK FISH pattern in NSCLC

<p>Comparison of Next-Generation Sequencing and Ventana Immunohistochemistry in Detecting ALK Rearrangements and Predicting the Efficacy of First-Line Crizotinib in Patients with Advanced Non-Small Cell Lung Cancer</p>

Gene rearrangement detection by next-generation sequencing in patients with non-small cell lung carcinoma

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