Detection of Rearrangements and Transcriptional Up-Regulation of ALK in FFPE Lung Cancer Specimens Using a Novel, Sensitive, Quantitative Reverse Transcription.

Slides:

Advertisements

Similar presentations

Naomi Fujioka, MD, Christopher A. French, MD, Michael J

Advertisements

ALK Protein Analysis by IHC Staining after Recent Regulatory Changes: A Comparison of Two Widely Used Approaches, Revision of the Literature, and a New.

Detection of ALK Gene Rearrangement in Non-small Cell Lung Cancer: A Comparison of Fluorescence In Situ Hybridization and Chromogenic In Situ Hybridization.

New Methods for ALK Status Diagnosis in Non–Small-Cell Lung Cancer: An Improved ALK Immunohistochemical Assay and a New, Brightfield, Dual ALK IHC–In.

Nicotine Induces Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor by α1 Nicotinic Acetylcholine Receptor–Mediated Activation in.

Rafal Dziadziuszko, MD, PhD, Anh T

ALK FISH and IHC: You Cannot Have One without the Other

Heterogeneity of Anaplastic Lymphoma Kinase Gene Rearrangement in Non–Small-Cell Lung Carcinomas: A Comparative Study Between Small Biopsy and Excision.

Jessica J. Lin, MD, Lauren L. Ritterhouse, MD, PhD, Siraj M

VAMP2–NRG1 Fusion Gene is a Novel Oncogenic Driver of Non–Small-Cell Lung Adenocarcinoma Yeonjoo Jung, PhD, Seunghui Yong, BS, Pora Kim, PhD, Hee-Young.

Identification and Characterization of ALK Kinase Splicing Isoforms in Non–Small-Cell Lung Cancer Lorena Lobo de Figueiredo-Pontes, MD, PhD, Daisy Wing-Sze.

Identification and Validation of Long Noncoding RNA Biomarkers in Human Non–Small- Cell Lung Carcinomas Hui Yu, MD, Qinghua Xu, PhD, Fang Liu, PhD, Xun.

Characterization of Fibroblast Growth Factor Receptor 1 in Small-Cell Lung Cancer Anish Thomas, MD, Jih-Hsiang Lee, MD, Zied Abdullaev, PhD, Kang-Seo.

FISH Analysis of Crizotinib Target Genes ROS1/ALK/MET in Malignant Mesothelioma Sandra Salvi, PhD, Serena Varesano, PhD, Simona Boccardo, PhD, Jean Louis.

Clinicopathologic Analysis of ROS1-Rearranged Non–Small-Cell Lung Cancer and Proposal of a Diagnostic Algorithm Heounjeong Go, MD, PhD, Dong-Wan Kim,

Large-Scale Screening and Molecular Characterization of EML4-ALK Fusion Variants in Archival Non–Small-Cell Lung Cancer Tumor Specimens Using Quantitative.

A Mass Spectrometry Assay to Simultaneously Analyze ROS1 and RET Fusion Gene Expression in Non–Small-Cell Lung Cancer Priyanga Wijesinghe, PhD, Gerold.

Increased ALK Gene Copy Number and Amplification are Frequent in Non-small Cell Lung Cancer Marta Salido, MSc, Lara Pijuan, MD, PhD, Luz Martínez-Avilés,

Frequent Coamplification and Cooperation between C-MYC and PVT1 Oncogenes Promote Malignant Pleural Mesothelioma Erick Riquelme, PhD, Milind B. Suraokar,

Thyroid Transcription Factor-1 Amplification and Expressions in Lung Adenocarcinoma Tissues and Pleural Effusions Predict Patient Survival and Prognosis

Optimization of Routine Testing for MET Exon 14 Splice Site Mutations in NSCLC Patients Clotilde Descarpentries, PharmD, Frédéric Leprêtre, PhD, Fabienne.

How I Do It—Optimal Methodology for Multidirectional Analysis of Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration Samples Takahiro Nakajima,

Bright-Field Dual-Color Chromogenic In Situ Hybridization for Diagnosing Echinoderm Microtubule-Associated Protein-Like 4-Anaplastic Lymphoma Kinase-Positive.

Interpretation of Anti-ALK Immunohistochemistry Results

Nicotine Induces Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor by α1 Nicotinic Acetylcholine Receptor–Mediated Activation in.

A Case of Lung Adenocarcinoma Resistant to Crizotinib Harboring a Novel EML4-ALK Variant, Exon 6 of EML4 Fused to Exon 18 of ALK Satoshi Anai, MD, Masafumi.

Inhibiting MDM2-p53 Interaction Suppresses Tumor Growth in Patient-Derived Non– Small Cell Lung Cancer Xenograft Models Josephine Hai, PhD, Shingo Sakashita,

PTPRF Expression as a Potential Prognostic/Predictive Marker for Treatment with Erlotinib in Non-Small-Cell Lung Cancer Denis Soulières, MD, Fred R.

A Novel Mass Spectrometry–Based Assay for Diagnosis of EML4-ALK–Positive Non– Small Cell Lung Cancer Kazuko Sakai, PhD, Isamu Okamoto, MD, PhD, Ken Takezawa,

Comprehensive Hybrid Capture–Based Next-Generation Sequencing Identifies a Double ALK Gene Fusion in a Patient Previously Identified to Be False-Negative.

ALK and NRG1 Fusions Coexist in a Patient with Signet Ring Cell Lung Adenocarcinoma Lucia Anna Muscarella, PhD, Domenico Trombetta, PhD, Federico Pio.

A Case of Squamous Cell Carcinoma Harboring an EML4-ALK Rearrangement that Was Unsuccessfully Treated with the ALK Inhibitor Alectinib Akihiro Tamiya,

Combined Use of ALK Immunohistochemistry and FISH for Optimal Detection of ALK- Rearranged Lung Adenocarcinomas Lynette M. Sholl, MD, Stanislawa Weremowicz,

Kelsie L. Thu, BSc, Raj Chari, PhD, William W

ALK/EML4 Fusion Gene May Be Found in Pure Squamous Carcinoma of the Lung Anna Caliò, MD, Alessia Nottegar, MD, Eliana Gilioli, MD, Emilio Bria, MD, Sara.

Tumor Cell Content for Selection of Molecular Techniques for T790M EGFR Mutation Detection in Non-small Cell Lung Cancer Nathalie Prim, Elisabeth Quoix,

Combinational Analysis of FISH and Immunohistochemistry Reveals Rare Genomic Events in ALK Fusion Patterns in NSCLC that Responds to Crizotinib Treatment

Reliability Assurance of Detection of EML4-ALK Rearrangement in Non–Small Cell Lung Cancer: The Results of Proficiency Testing in China Yulong Li, MD,

ALK Rearrangement Detected in a Focus of Pulmonary Atypical Adenomatous Hyperplasia Filippo Lococo, MD, Alessandra Bisagni, MD, Maria Cecilia Mengoli,

Searching for ROS1 Rearrangements in Lung Cancer by Fluorescent In Situ Hybridization: The Importance of Probe Design Arnaud Uguen, MD, Pascale Marcorelles,

A Novel EML4-ALK Variant: Exon 6 of EML4 Fused to Exon 19 of ALK

Parallel FISH and Immunohistochemical Studies of ALK Status in 3244 Non–Small-Cell Lung Cancers Reveal Major Discordances Florian Cabillic, PharMD, PhD,

Detection of ALK Rearrangement by Immunohistochemistry in Lung Adenocarcinoma and the Identification of a Novel EML4-ALK Variant Ka-Fai To, MBChB, Joanna.

Identification of a Novel HIP1-ALK Fusion Variant in Non–Small-Cell Lung Cancer (NSCLC) and Discovery of ALK I1171 (I1171N/S) Mutations in Two ALK-Rearranged.

ALK Testing in Lung Adenocarcinoma: Technical Aspects to Improve FISH Evaluation in Daily Practice Vittoria Martin, PhD, Barbara Bernasconi, PhD, Elisabetta.

Heterogeneity of Genetic Changes Associated with Acquired Crizotinib Resistance in ALK-Rearranged Lung Cancer Soyeon Kim, PhD, Tae Min Kim, MD, PhD,

Erratum Journal of Thoracic Oncology

Arnaud Uguen, MD, PhD Journal of Thoracic Oncology

High MET Receptor Expression But Not Gene Amplification in ALK 2p23 Rearrangement Positive Non–Small-Cell Lung Cancer Yan Feng, MD, Eugen C. Minca, MD,

Nicotinamide Phosphoribosyltransferase: A Potent Therapeutic Target in Non-small Cell Lung Cancer with Epidermal Growth Factor Receptor-Gene Mutation

Detection of ALK-Positive Non–Small-Cell Lung Cancers on Cytological Specimens: High Accuracy of Immunocytochemistry with the 5A4 Clone Spasenija Savic,

Clinicopathologic Characteristics and Outcomes of Patients with Anaplastic Lymphoma Kinase-Positive Advanced Pulmonary Adenocarcinoma: Suggestion for.

Daniel B. Costa, MD, PhD, Susumu Kobayashi, MD, PhD

EML4-ALK Fusion Detected by RT-PCR Confers Similar Response to Crizotinib as Detected by FISH in Patients with Advanced Non-Small-Cell Lung Cancer Yan.

Atypical Negative ALK Break-Apart FISH Harboring a Crizotinib-Responsive ALK Rearrangement in Non–Small-Cell Lung Cancer Shengxiang Ren, MD, PhD, Fred.

A Novel Fusion of TPR and ALK in Lung Adenocarcinoma

Toni-Maree Rogers, BMLS, Prudence A

Immunohistochemistry is a Reliable Screening Tool for Identification of ALK Rearrangement in Non–Small-Cell Lung Carcinoma and is Antibody Dependent

Driven by Mutations: The Predictive Value of Mutation Subtype in EGFR-Mutated Non– Small Cell Lung Cancer Emily Castellanos, MD, Emily Feld, MD, Leora.

HIP1–ALK, A Novel ALK Fusion Variant that Responds to Crizotinib

Concomitant Epidermal Growth Factor Receptor Mutation and EML4-ALK Fusion in a Patient with Multifocal Lung Adenocarcinomas Jun Fan, MD Journal of Thoracic.

Clinical Implications of Variant ALK FISH Rearrangement Patterns

Highly Sensitive Detection of EGFR T790M Mutation in Pre-TKI Specimens of EGFR- Mutated NSCLC: In Cis, In Trans, or a Different Clone? Alvaro Leone, BSc.D

Good Response to Gefitinib in Lung Adenocarcinoma Harboring Coexisting EML4-ALK Fusion Gene and EGFR Mutation Yao-Wen Kuo, MD, Shang-Gin Wu, MD, Chao-Chi.

An International Interpretation Study Using the ALK IHC Antibody D5F3 and a Sensitive Detection Kit Demonstrates High Concordance between ALK IHC and.

A Dramatic Response to Crizotinib in a Non–Small-Cell Lung Cancer Patient with IHC- Positive and FISH-Negative ALK Jong-Mu Sun, MD, PhD, Yoon-La Choi,

ALK Status Testing in Non–Small-Cell Lung Carcinoma by FISH on ThinPrep Slides with Cytology Material Eugen C. Minca, MD, PhD, Christopher P. Lanigan,

Waxing and Waning of MET Amplification in EGFR-Mutated NSCLC in Response to the Presence and Absence of Erlotinib Selection Pressure Joel P. Womack,

ALK Gene Rearrangements: A New Therapeutic Target in a Molecularly Defined Subset of Non-small Cell Lung Cancer Benjamin Solomon, MBBS, PhD, Marileila.

The Current Role of Whole Brain Radiation Therapy in Non–Small Cell Lung Cancer Patients Gokoulakrichenane Loganadane, MD, Lizza Hendriks, MD, PhD, Cécile.

Presentation transcript:

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 Kim Gruber, MSc, Heike Horn, PhD, Jörg Kalla, MD, Peter Fritz, MD, Andreas Rosenwald, MD, Martin Kohlhäufl, MD, Godehard Friedel, MD, Matthias Schwab, MD, German Ott, MD, Claudia Kalla, PhD Journal of Thoracic Oncology Volume 9, Issue 3, Pages 307-315 (March 2014) DOI: 10.1097/JTO.0000000000000068 Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 ALK mRNA expression in NSCLC. A, Genomic structure of ALK according to NM_004304. Amplicons of the qRT-PCR assay are indicated by boxes: red box, ALK 5′ portion; blue box, ALK 3′ portion. B, qRT-PCR analysis of ALK was performed relative to the PGK1 housekeeping gene and calculated relative to the average value in cell line RH30, which was arbitrarily defined as 1. The horizontal dashed line indicates the cutoff value for altered ALK expression (0.3); red bars, ALK 5′portion; blue bars, ALK 3′ portion. The asterisks point to tumors expressing full-length transcripts. ALK, anaplastic large-cell lymphoma kinase; NSCLC, non–small-cell lung cancer; PGK1, phophoglycerokinase 1; qRT-PCR, quantitative reverse transcription polymerase chain reaction. Journal of Thoracic Oncology 2014 9, 307-315DOI: (10.1097/JTO.0000000000000068) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Comparative analysis of ALK by qRT-PCR measuring the expression of the 5′ and the 3′ portion of ALK transcripts (left), FISH using probes flanking the ALK gene (middle) and immunostaining for ALK protein (right). A and B, ALK-rearranged tumors expressing the ALK 3′ portion; (C) tumor expressing full-length ALK transcript in the absence of gene rearrangement. Gray bars, ALK transcript 5′portion; dark gray bars, ALK transcript 3′ portion; arrows, split FISH signals characteristic of an ALK rearrangement; scale bar, 200 μm. ALK, anaplastic large-cell lymphoma kinase; qRT-PCR, quantitative reverse transcription polymerase chain reaction; NSCLC, non–small-cell lung cancer; FISH, fluorescence in situ hybridization. Journal of Thoracic Oncology 2014 9, 307-315DOI: (10.1097/JTO.0000000000000068) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 ALK mutation and full-length transcript expression in NSCLC #25. A, Tumor and nonmalignant cells harbor the heterozygous nucleotide substitution 3659C>A, resulting in the replacement of a serine by a tyrosine within the kinase domain (S1220Y). B, Full-length ALK transcript was expressed in tumor, but not in nonmalignant cells, as detected by qRT-PCR. Light gray bars, ALK 5′portion; dark gray bars, ALK 3′ portion. ALK, anaplastic large-cell lymphoma kinase; qRT-PCR, quantitative reverse transcription polymerase chain reaction; NSCLC, non–small-cell lung cancer. Journal of Thoracic Oncology 2014 9, 307-315DOI: (10.1097/JTO.0000000000000068) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions