Cytology specimens offer an effective alternative to formalin‐fixed tissue as demonstrated by novel automated detection for ALK break‐apart FISH testing and immunohistochemistry in lung adenocarcinoma

Rosenblum, Frida; Hutchinson, Lloyd; Garver, Joann; Woda, Bruce A.; Cosar, Ediz F.; Kurian, Elizabeth

doi:10.1002/cncy.21467

Cited by 30 publications

(43 citation statements)

References 51 publications

(75 reference statements)

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“…Superimposed signals indicate ALK wild-type status, while inversion and rearrangements generate signals that can be identified as split or isolated (36). This technique works appropriately either on archival paraffin-embedded tumor specimens and well-prepared, mono-layer, non-bloody smeared cytology (37,38). A cut-off of >15% rearranged tumor cells, counting at least 50 tumor cell nuclei, has been decided to quote ALK fusions by FISH (Figure 1) (26).…”

Section: Fishmentioning

confidence: 99%

Tackling ALK in non-small cell lung cancer: the role of novel inhibitors

Facchinetti¹,

Tiseo²,

Maïo³

et al. 2016

Transl. Lung Cancer Res.

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Abstract:Crizotinib is an oral inhibitor of anaplastic lymphoma kinase (ALK) with remarkable clinical activity in patients suffering from ALK-rearranged non-small cell lung cancer (NSCLC), accounting to its superiority compared to chemotherapy. Unfortunately, virtually all ALK-rearranged tumors acquire resistance to crizotinib, frequently within one year since the treatment initiation. To date, therapeutic strategies to overcome crizotinib resistance have focused on the use of more potent and structurally different compounds. Second-generation ALK inhibitors such as ceritinib (LDK378), alectinib (CH5424802/ RO5424802) and brigatinib (AP26113) have shown relevant clinical activity, consequently fostering their rapid clinical development and their approval by health agencies. The third-generation inhibitor lorlatinib (PF-06463922), selectively active against ALK and ROS1, harbors impressive biological potency; its efficacy in reversing resistance to crizotinib and to other ALK inhibitors is being proven by early clinical trials. The NTRK1-3 and ROS1 inhibitor entrectinib (RXDX-101) has been reported to act against NSCLC harboring ALK fusion proteins too. Despite the quick development of these novel agents, several issues remain to be discussed in the treatment of patients suffering from ALK-rearranged NSCLC. This position paper will discuss the development, the current evidence and approvals, as long as the future perspectives of new ALK inhibitors beyond crizotinib. Clinical behaviors of ALK-rearranged NSCLC vary significantly among patients and differential molecular events responsible of crizotinib resistance account for the most important quote of this heterogeneity. The precious availability of a wide range of active anti-ALK compounds should be approached in a critical and careful perspective, in order to develop treatment strategies tailored on the disease evolution of every single patient.

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Section: Fishmentioning

confidence: 99%

Tackling ALK in non-small cell lung cancer: the role of novel inhibitors

Facchinetti¹,

Tiseo²,

Maïo³

et al. 2016

Transl. Lung Cancer Res.

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“…4,5 Immunoperoxidase testing and fluorescence in situ hybridization (FISH) can also be applied to cytology specimens. [6][7][8][9] Although cytology specimen processing is not standardized across laboratories, [10][11][12] the advantage of this lack of standardization is that investigators can make use of an array of techniques and find the one most conducive to their methods. Direct smears, cell monolayers, and cell block sections can all be used for IPOX and molecular tests including next-generation sequencing.…”

Section: Cytology Specimensmentioning

confidence: 99%

“…45,46 Although the first ALK antibody clones were unreliable, the subsequently developed clones D5F3 (Cell Signaling Technology) and 5A4 (Novocastra, Leica Biosystems, Buffalo Grove, Illinois) have a reported sensitivity of 100% and specificity of 99% in biopsy specimens, with excellent interobserver agreement and concordance with FISH results. 6,7,17,26,47,48 Clone 5A4, which is as reliable as D5F3 in the detection of ALK rearrangement, is used more commonly in Europe. 17,49 The performance of ALK IPOX in cytology specimens (formalin-fixed cell blocks) thus far is similar to that in surgical specimens.…”

Section: Alk Antibodiesmentioning

confidence: 99%

“…Additional validation studies are needed. [6][7][8]16 The recently FDA-approved IPOX assay for ALK rearrangement using the D5F3 clone qualitatively detects ALK rearrangement in NSCLC as a companion diagnostic test to select patients for crizotinib treatment. 26 The assay is FDA approved for use on formalin-fixed, paraffin-embedded NSCLC tissue using Ventana's rabbit monoclonal antibody (licensed from Cell Signaling Technology), OptiView Amplification Kit, OptiView DAB IHC Detection Kit, and BenchMark XT automated slide staining instrument.…”

Section: Alk Antibodiesmentioning

confidence: 99%

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Lung Carcinoma Predictive Biomarker Testing by Immunoperoxidase Stains in Cytology and Small Biopsy Specimens: Advantages and Limitations

Zhou

Moreira²

2016

Archives of Pathology &Amp; Laboratory Medicine

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Context.-In the burgeoning era of molecular genomics, immunoperoxidase (IPOX) testing grows increasingly relevant as an efficient and effective molecular screening tool. Patients with lung carcinoma may especially benefit from the use of IPOX because most lung carcinomas are inoperable at diagnosis and only diagnosed by small tissue biopsy or fine-needle sampling. When such small specimens are at times inadequate for molecular testing, positive IPOX results still provide actionable information.Objective.-To describe the benefits and pitfalls of IPOX in the detection of biomarkers in lung carcinoma cytology specimens and small biopsies by summarizing the currently available commercial antibodies, preanalytic variables, and analytic considerations.Data Sources.-PubMed.Conclusions.-Commercial antibodies exist for IPOX detection of aberrant protein expression due to EGFR L858R mutation, EGFR E746_A750 deletion, ALK rearrangement, ROS1 rearrangement, and BRAF V600E mutation, as well as PD-L1 expression in tumor cells. Automated IPOX protocols for ALK and PD-L1 detection were recently approved by the Food and Drug Administration as companion diagnostics for targeted therapies, but consistent interpretive criteria remain to be elucidated, and such protocols do not yet exist for other biomarkers. The inclusion of cytology specimens in clinical trials would expand patients' access to testing and treatment, yet there is a scarcity of clinical trial data regarding the application of IPOX to cytology, which can be attributed to trial designers' lack of familiarity with the advantages and limitations of cytology. The content of this review may be used to inform clinical trial design and advance IPOX validation studies.

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“…The short answer is that any cytology preparation (eg, smears, monolayer preparations, or cell blocks fixed by various means) can produce a valid molecular test result, and a key consideration is to use the preparation with sufficient numbers of tumor cells (Table ) . The devil is in the details, however, and the many publications that have established the suitability of cytology samples for molecular diagnosis continue to raise new questions.…”

Section: Cytologic Platforms For Molecular Testingmentioning

confidence: 99%

Technical and US regulatory issues in triaging material for the molecular laboratory

Fischer

Hutchinson

2016

Cancer Cytopathology

Self Cite

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We recently performed a fine-needle aspiration (FNA) on a patient aged 28 years, a father of 2 children, with a deep neck mass. Smears on site looked like melanoma, but there was no history. Immunohistochemistry and potentially molecular testing would be needed. For example, it could be useful to analyze the gene BRAF, coding for the serine/threonine protein kinase B-Raf, to identify mutations that confer survival benefit to melanoma patients treated with kinase inhibitors.1 Scenarios like this are playing out with increasing frequency in cytology.What exactly is the best way to triage material for the molecular laboratory?The short answer is that any cytology preparation (eg, smears, monolayer preparations, or cell blocks fixed by various means) can produce a valid molecular test result, and a key consideration is to use the preparation with sufficient numbers of tumor cells 2 (Table 1). [3][4][5][6][7][8][9][10][11][12][13] The devil is in the details, however, and the many publications that have established the suitability of cytology samples for molecular diagnosis continue to raise new questions. Beyond the scientific questions, molecular laboratories in the United States need to stay compliant with the Clinical Laboratory Improvement Act (CLIA). More challenging still could be the proposed new US Food and Drug Administration (FDA) regulations that would require laboratory-developed tests (LDTs) to be granted FDA approvals. 14 These regulations, if enacted as described, threaten the ability of cytology to help patients who need molecular testing. In this article, we have restricted attention only to BRAF mutation testing as an example of how technologies and the regulatory climate affect the ability of cytology laboratories to provide a valuable service for patients. Three Factors That Affect the Suitability of Cytology Samples for Molecular TestingThe 3 key factors that affect the validity of a sequence-based molecular assay are the proportion of tumor cells in the sample, DNA quality, and DNA quantity. A low proportion of tumor cells could yield a dangerous falsenegative result, rendering a mutation undetectable, even with the most robust assays and controls. An advantage of cytology samples compared with surgical biopsies is that they frequently have a higher proportion of carcinoma cells. Newer microbiopsy devices may help to improve the procurement of heavily cross-linked sarcomas or desmoplastic carcinomas. The FDA-approved Roche Cobas BRAF test (Roche Molecular Systems, Inc, Pleasanton, Calif) detects mutant valine codon 600 (V600) alleles when tumor cells comprise 15% of total cells. It is not trivial to estimate the percentage of tumor cells, and data on the accuracy of pathologists' estimate of tumor cellularity have been limited to paraffin sections. By using an objective counting method as a gold standard, pathologists in

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Cytology specimens offer an effective alternative to formalin‐fixed tissue as demonstrated by novel automated detection for ALK break‐apart FISH testing and immunohistochemistry in lung adenocarcinoma

Cited by 30 publications

References 51 publications

Tackling ALK in non-small cell lung cancer: the role of novel inhibitors

Tackling ALK in non-small cell lung cancer: the role of novel inhibitors

Lung Carcinoma Predictive Biomarker Testing by Immunoperoxidase Stains in Cytology and Small Biopsy Specimens: Advantages and Limitations

Technical and US regulatory issues in triaging material for the molecular laboratory

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