Undifferentiated Small Round Cell Sarcomas with Rare EWS Gene Fusions

Slides:

Advertisements

Similar presentations

Margaret L. Gulley, Thomas C. Shea, Yuri Fedoriw

Advertisements

Establishment and characterization of a new human acute myelocytic leukemia cell line SH-2 with a loss of Y chromosome, a derivative chromosome 16 resulting.

A Genome-Wide High-Resolution Array-CGH Analysis of Cutaneous Melanoma and Comparison of Array-CGH to FISH in Diagnostic Evaluation Lu Wang, Mamta Rao,

Volume 204, Issue 1, Pages (January 2011)

Detection of TMPRSS2-ETS Fusions by a Multiprobe Fluorescence in Situ Hybridization Assay for the Early Diagnosis of Prostate Cancer Qi-Peng Sun, Liao-Yuan.

Droplet Digital PCR for Absolute Quantification of EML4-ALK Gene Rearrangement in Lung Adenocarcinoma Qiushi Wang, Xin Yang, Yong He, Qiang Ma, Li Lin,

Tracy I. George, Joanna E. Wrede, Charles D. Bangs, Athena M

Molecular Diagnosis of Clear Cell Sarcoma

Detection of TMPRSS2-ETS Fusions by a Multiprobe Fluorescence in Situ Hybridization Assay for the Early Diagnosis of Prostate Cancer Qi-Peng Sun, Liao-Yuan.

Margaret L. Gulley, Thomas C. Shea, Yuri Fedoriw

A Case of FIP1L1-PDGFRA-Positive Chronic Eosinophilic Leukemia with a Rare FIP1L1 Breakpoint Frédéric Lambert, Pierre Heimann, Christian Herens, Alain.

A Single-Tube Multiplexed Assay for Detecting ALK, ROS1, and RET Fusions in Lung Cancer Maruja E. Lira, Yoon-La Choi, Sun Min Lim, Shibing Deng, Donghui.

Mutation Spectrum of the Survival of Motor Neuron 1 and Functional Analysis of Variants in Chinese Spinal Muscular Atrophy Yu-jin Qu, Jin-li Bai, Yan-yan.

Carmo Martins, Branca Cavaco, Giovanni Tonon, Frederic J

Β-Glucuronidase Is an Optimal Normalization Control Gene for Molecular Monitoring of Chronic Myelogenous Leukemia Joong Won Lee, Qiaofang Chen, Daniel.

Jonathan A. Schumacher, Stephen D. Jenson, Kojo S. J

Tony L. Ng, Maureen J. O'Sullivan, Catherine J

Betaine, Dimethyl Sulfoxide, and 7-Deaza-dGTP, a Powerful Mixture for Amplification of GC-Rich DNA Sequences Marco Musso, Renata Bocciardi, Sara Parodi,

FISH Analysis for the Detection of Lymphoma-Associated Chromosomal Abnormalities in Routine Paraffin-Embedded Tissue Roland A. Ventura, Jose I. Martin-Subero,

Philippe Szankasi, Mohamed Jama, David W. Bahler

Jennelle C. Hodge, Patrick P. Bedroske, Kathryn E. Pearce, William R

Down Syndrome and Malignancies: A Unique Clinical Relationship

Hou-Sung Jung, Gregory J. Tsongalis, Joel A. Lefferts

Fluorescence in Situ Hybridization Analysis of Immunoglobulin Heavy Chain Translocations in Plasma Cell Myeloma Using Intact Paraffin Sections and Simultaneous.

Molecular Diagnosis in Ewing Family Tumors

A Tumor Sorting Protocol that Enables Enrichment of Pancreatic Adenocarcinoma Cells and Facilitation of Genetic Analyses Zachary S. Boyd, Rajiv Raja,

Structural and Functional Mutations of the Perlecan Gene Cause Schwartz-Jampel Syndrome, with Myotonic Myopathy and Chondrodysplasia Eri Arikawa-Hirasawa,

A Novel Alu-Like Element Rearranged in the Dystrophin Gene Causes a Splicing Mutation in a Family with X-Linked Dilated Cardiomyopathy Alessandra Ferlini,

Assessment of EGFR Mutation Status in Lung Adenocarcinoma by Immunohistochemistry Using Antibodies Specific to the Two Major Forms of Mutant EGFR Marie.

Fusion of TEL, the ETS-Variant Gene 6 (ETV6), to the Receptor-Associated Kinase JAK2 as a Result of t(9; 12) in a Lymphoid and t(9; 15; 12) in a Myeloid.

Hepatosplenic and Subcutaneous Panniculitis-Like γ/δ T Cell Lymphomas Are Derived from Different Vδ Subsets of γ/δ T Lymphocytes Grzegorz K. Przybylski,

SeqSharp The Journal of Molecular Diagnostics

A Genome-Wide High-Resolution Array-CGH Analysis of Cutaneous Melanoma and Comparison of Array-CGH to FISH in Diagnostic Evaluation Lu Wang, Mamta Rao,

Janice M. Spence, Paul G. Rothberg, Nancy Wang, W. Richard Burack

Double Heterozygosity for a RET Substitution Interfering with Splicing and an EDNRB Missense Mutation in Hirschsprung Disease Alberto Auricchio, Paola.

Molecular Characterization of WFS1 in Patients with Wolfram Syndrome

Analysis of Rare APC Variants at the mRNA Level

Inconspicuous Insertion 22;12 in Myxoid/Round Cell Liposarcoma Accompanied by the Secondary Structural Abnormality der(16)t(1;16) Nathan C. Birch, Cristina.

Diagnostic Utility of Molecular Investigation in Extraskeletal Myxoid Chondrosarcoma Stefania Benini, Stefania Cocchi, Gabriella Gamberi, Giovanna Magagnoli,

The MECT1-MAML2 Gene Fusion and Benign Warthin's Tumor

Novel Fluorescent Ligase Detection Reaction and Flow Cytometric Analysis of SYT-SSX Fusions in Synovial Sarcoma Robyn Gaffney, Artemis Chakerian, John.

Activation of a Cryptic Splice Site of PTEN and Loss of Heterozygosity in Benign Skin Lesions in Cowden Disease Stephen J. Meltzer, Manfred Wolter Journal.

Molecular Monitoring of Chronic Myelogenous Leukemia

A Homozygous Nonsense Mutation in Type XVII Collagen Gene (COL17A1) Uncovers an Alternatively Spliced mRNA Accounting for an Unusually Mild Form of Non-Herlitz.

Daynna J. Wolff, Adam Bagg, Linda D. Cooley, Gordon W. Dewald, Betsy A

A Presenilin-1 Truncating Mutation Is Present in Two Cases with Autopsy-Confirmed Early-Onset Alzheimer Disease Carolyn Tysoe, Joanne Whittaker, John.

Defining Ploidy-Specific Thresholds in Array Comparative Genomic Hybridization to Improve the Sensitivity of Detection of Single Copy Alterations in Cell.

Molecular Cytogenetic Analyses of Immunoglobulin Loci in Nodular Lymphocyte Predominant Hodgkin's Lymphoma Reveal a Recurrent IGH-BCL6 Juxtaposition

N. Scott Reading, Stephen D. Jenson, Jeffrey K. Smith, Megan S

Evaluating the Effect of Unclassified Variants Identified in MMR Genes Using Phenotypic Features, Bioinformatics Prediction, and RNA Assays Lucia Pérez-Cabornero,

A Novel Type of EWS-CHOP Fusion Gene in Two Cases of Myxoid Liposarcoma Taisuke Hosaka, Yasuaki Nakashima, Katsuyuki Kusuzaki, Hiroaki Murata, Tomitaka.

Array Comparative Genomic Hybridization Detects Chromosomal Abnormalities in Hematological Cancers That Are Not Detected by Conventional Cytogenetics

Jianbo Song, Danielle Mercer, Xiaofeng Hu, Henry Liu, Marilyn M. Li

The Detection of t(14;18) in Archival Lymph Nodes

Standard Mutation Nomenclature in Molecular Diagnostics

Deletion of the Cytoplasmatic Domain of BP180/Collagen XVII Causes a Phenotype with Predominant Features of Epidermolysis Bullosa Simplex Marcel Huber,

Amplification Refractory Mutation System, a Highly Sensitive and Simple Polymerase Chain Reaction Assay, for the Detection of JAK2 V617F Mutation in Chronic.

Feras M. Hantash, Arlene Rebuyon, Mei Peng, Joy B

Accurate, simple, and inexpensive assays to diagnose F8 gene inversion mutations in hemophilia A patients and carriers by Debargh Dutta, Devi Gunasekera,

Compound Heterozygosity for Novel Splice Site Mutations in the BPAG2/COL17A1 Gene Underlies Generalized Atrophic Benign Epidermolysis Bullosa Leena Pulkkinen,

Comprehensive Characterization of a Novel Intronic Pseudo-Exon Inserted within an e14/a2 BCR-ABL Rearrangement in a Patient with Chronic Myeloid Leukemia

Annemieke Aartsma-Rus, Anneke A. M. Janson, Wendy E

Molecular Diagnosis of Mast Cell Disorders

Jie Hu, Malini Sathanoori, Sally J. Kochmar, Urvashi Surti

KIT Gene Deletions at the Intron 10−Exon 11 Boundary in GI Stromal Tumors Christopher L. Corless, Laura McGreevey, Ajia Town, Arin Schroeder, Troy Bainbridge,

Fluorescence In Situ Hybridization Identifies Cryptic t(16;16)(p13;q22) Masked By del(16)(q22) in a Case of AML-M4 Eo Shakil H. Merchant, Skip Haines,

Genetic Characterization of a Human Skin Carcinoma Progression Model: from Primary Tumor to Metastasis Susanne Popp, Stefan Waltering, Petra Boukamp

Maureen J. O'Sullivan, Peter A. Humphrey, Louis P. Dehner, John D

Volume 3, Issue 3, Pages (March 1999)

Exon Skipping in IVD RNA Processing in Isovaleric Acidemia Caused by Point Mutations in the Coding Region of the IVD Gene Jerry Vockley, Peter K. Rogan,

Presentation transcript:

Undifferentiated Small Round Cell Sarcomas with Rare EWS Gene Fusions Lu Wang, Rohit Bhargava, Tao Zheng, Leonard Wexler, Margaret H. Collins, Diane Roulston, Marc Ladanyi The Journal of Molecular Diagnostics Volume 9, Issue 4, Pages 498-509 (September 2007) DOI: 10.2353/jmoldx.2007.070053 Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Patient A. A: H&E-stained section showing characteristic small round blue cell tumor morphology. B: Diffuse and strong positivity for CD99 (O13 antibody). C: Partial karyotype showing a t(7;22) translocation. Arrows indicate the derivative chromosomes. The Journal of Molecular Diagnostics 2007 9, 498-509DOI: (10.2353/jmoldx.2007.070053) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Patient B. Undifferentiated tumor showing areas of spindling in a specimen obtained after neoadjuvant chemotherapy. The Journal of Molecular Diagnostics 2007 9, 498-509DOI: (10.2353/jmoldx.2007.070053) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Patient C. A: H&E-stained section showing small round blue cell tumor with crush artifact. B: Immunostain for CD99 showing diffuse and strong staining. C: CAM5.2 immunostain showing dot-like positivity for cytokeratin in the tumor cells. The Journal of Molecular Diagnostics 2007 9, 498-509DOI: (10.2353/jmoldx.2007.070053) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 Patient D. A: H&E-stained section showing small round blue cell tumor with high mitotic rate. B: Immunostain for CD99 showing focal weak staining. C: BCL2 immunostain showing strong diffuse positivity in the tumor cells. The Journal of Molecular Diagnostics 2007 9, 498-509DOI: (10.2353/jmoldx.2007.070053) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 5 Patient D. A: Partial karyotype showing t(2;22)(q31;q12). Arrows indicate the derivative chromosomes. B: FISH using dual-color, break-apart EWS probe on a metaphase preparation. The small arrow shows the normal fusion signal (red and green signals together) on the intact chromosome 22; the large arrow indicates the der(22)t(2;22) with only red signal (EWS centromeric probe); the midsize arrow indicates the der(2)t(2;22) with the green signal (EWS telomeric probe). C: Same FISH on interphase nuclei showing one normal fusion signal and separation of the other red and green signals. The Journal of Molecular Diagnostics 2007 9, 498-509DOI: (10.2353/jmoldx.2007.070053) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 6 Patient D. A: Partial sequence of the type I EWS-SP3 fusion transcript is an in-frame junction of EWS exon 7 with SP3 exon 6 (see Results for details). The translated protein sequence is shown below, with the codon at the junction in red. B: Partial sequence of the type II EWS-SP3 transcript. The transcript has a full open reading frame and is a fusion of EWS exon 8 to SP3 exon 6 with an intervening cryptic exon. The arrowhead indicates the extra T (see Results for details). The translated protein sequence is shown below, with the codons at the junctions in red. C: Partial genomic sequence of SP3. The intron sequence is in lower case and the exon sequence is in upper case. The portion of SP3 exon 6 coding for the zinc finger is underlined. The two cryptic splice acceptor sites in SP3 exon 6 are shown in green (used in type I fusion transcript) and red (used type II fusion transcript), respectively. The Journal of Molecular Diagnostics 2007 9, 498-509DOI: (10.2353/jmoldx.2007.070053) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 7 Patient D: RT-PCR detection of EWS-SP3 fusion. The presence of both type I and type II fusion transcripts was confirmed by RT-PCR using an EWS primer (sequence: 5′-CTGGATCCTACAGCCAAGCTCCAAGTC-3′) and an SP3 reverse primer (sequence: 5′-CAGAATGCCAACGCAGATGA-3′). Lane ptD is the RT-PCR product obtained from tumor RNA. Band A is 120 bp, and band B is 375 bp. Sequencing of A and B confirmed that A represents the type I EWS-SP3 fusion transcript, whereas B represents the type II fusion, both of which are in-frame (see Figure 6). Sequencing of band C revealed an out-of-frame EWS-SP3 fusion transcript (sequence data not shown). Lane neg is the no RNA negative control. Size marker (lane M) is ϕ X174/HaeIII (Invitrogen). The Journal of Molecular Diagnostics 2007 9, 498-509DOI: (10.2353/jmoldx.2007.070053) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 8 Schematic diagrams of EWS, SP3, and the predicted EWS-SP3 protein. NTD, amino-terminal domain; AD, transactivation domain; ID, inhibitory domain; zinc finger, zinc finger DNA-binding domain. The Journal of Molecular Diagnostics 2007 9, 498-509DOI: (10.2353/jmoldx.2007.070053) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions