F.I.S.H. in PATHOLOGY; Applications,Answers..and More Allan Kennedy Lead Biomedical Scientist (Molecular Pathology) Glasgow Royal Infirmary.

Slides:

Advertisements

Similar presentations

GT training- A GOSH experience Sumera Ghani Great Ormond St Hospital London Paediatric Malignancy Unit.

Advertisements

MYC/BCL2 protein coexpression contributes inferior outcome in DLBCL

Niko Bla ž evi ć Mentor: A. Ž mega č Horvat.  The process of transformation from a normal cell to a cancerous one  Synonym: neoplasia Carcin ogenes.

Fluorescence in situ Hybridization

NYU SCHOOL OF MEDICINE CYTOGENETICS LECTURE 2

Molecular Pathology – Cell cycle Dr. Leonard Da Silva Senior Lecturer Molecular & Cellular Pathology.

Non-Hodgkin Lymphoma Showing Abnormalities of c-myc Including Dual Translocations Involving c-myc and Bcl-2: A Clinicopathologic Study R Jastania, V Kukreti,

Generation of Oncogenes The major ways of generating oncogenes: Proliferation stimulating gene acquired by a virus. Proviral insertional mutagenesis places.

The Loss of the Cell Cycle Control in Cancer

Cellular oncogenes.

Chapter 14 Molecular Oncology.

Burkitt’s Lymphoma Catherine Hanson Richard Hildreth David Duke Kelly Lewis Ray Lewis.

Soft Tissue Tumors Academy of Pathology and Laboratory Medicine of Puerto Rico April 2013 Bruce Horten, M.D. Medical Director Integrated Oncology, New.

Fluorescent In Situ Hybridization (FISH) to Identify Genetic Changes in Fine Needle Biopsy of Lung Lesions Prepared by Jin Jen NCI.

S.Shetty 1, N. Uddin 1,2,K.B. Geiersbach 1, S.T. South 1,2. 1)University of Utah, School of Medicine, Department of Pathology, Salt lake City, UT and Cytogenetics/Molecular.

Extending FISH Analysis of Paediatric Tumours

Bone and soft tissue tumors Imad Fadl-Elmula Al Neelain University.

Chapter 4 Cellular Oncogenes ~ Mar 22, 2007.

Normal haemopoiesis. ABNORMALITIES IN THE HEMOPOIETIC SYSTEM CAN LEAD TO HEMOGLOBINOPATHIES HEMOPHILIA DEFECTS IN HEMOSTASIS/THROMBOSIS HEMATOLOGICAL.

YUEMIN DING Neuro-oncology Group Department of Molecular Neuroscience

FISHing in Pathology Gopalrao Velagaleti, Ph.D Director, Cytogenetics Laboratory.

Sarcomas Perspectives and Background. Sarcomas: Themes Sarcomas are a heterogeneous collection of diseases and families of diseases –Individual diseases/families.

Chromosomal abnormalities and tumors Seminar No 524, course: Cells and tissues development.

Genetics of Cancer Lecture 34.

Non Hodgkin”s Lymphoma -- Histology appearance -- Cell of orgin -- Immunophenotype -- Molecular biology -- Clinical featres -- Prognosis -- Out-come of.

Helicobacter pylori and Gastric Lymphoma

CONTRIBUTION OF CHROMOSOME BANDING AND MOLECULAR CYTOGENETIC ANALYSES FOR THE DIAGNOSIS OF SOFT TISSUE AND BONE TUMORS OVER A 6-YEAR-PERIOD Manuel Teixeira,

GENETIC BIOMARKERS.

Genetic Testing in Sarcoma: Current practice and future perspectives

Neoplasia lecture4 Dr Heyam Awad FRCPath.

Aberrant Calreticulin Expression Is Involved in the Dedifferentiation of Dedifferentiated Liposarcoma Masanori Hisaoka, Atsuji Matsuyama, Mitsuhiro Nakamoto

Fluorescent In Situ Hybridization (FISH) Assay

Whole-Genome Scanning by Array Comparative Genomic Hybridization as a Clinical Tool for Risk Assessment in Chronic Lymphocytic Leukemia Shelly R. Gunn,

Volume 52, Issue 1, Pages (July 2007)

FISH Panel for Leukemic CTCL

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

Focus on lymphomas Cancer Cell

Harvey A. Greisman, Noah G. Hoffman, Hye Son Yi

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

A distinctive subtype of t(14;18)-negative nodal follicular non-Hodgkin lymphoma characterized by a predominantly diffuse growth pattern and deletions.

Multiplex Ligation-Dependent Probe Amplification

Detection of Genetic Alterations by ImmunoFISH Analysis of Whole Cells Extracted from Routine Biopsy Material Göran Mattsson, Soo Yong Tan, David J.P.

A New Role for Hypoxia in Tumor Progression

Whole-Genome Scanning by Array Comparative Genomic Hybridization as a Clinical Tool for Risk Assessment in Chronic Lymphocytic Leukemia Shelly R. Gunn,

Molecular Diagnostic Approach to Non-Hodgkin's Lymphoma

Molecular Diagnosis in Ewing Family Tumors

Undifferentiated Small Round Cell Sarcomas with Rare EWS Gene Fusions

Customized Oligonucleotide Array-Based Comparative Genomic Hybridization as a Clinical Assay for Genomic Profiling of Chronic Lymphocytic Leukemia Rachel.

Molecular Analysis of Gene Fusions in Bone and Soft Tissue Tumors by Anchored Multiplex PCR–Based Targeted Next-Generation Sequencing Suk Wai Lam, Anne-Marie.

The Development of a Multitarget, Multicolor Fluorescence in Situ Hybridization Assay for the Detection of Urothelial Carcinoma in Urine Irina A. Sokolova,

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,

Caught in transformation

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

Multiplex Ligation-Dependent Probe Amplification Versus Multiprobe Fluorescence in Situ Hybridization To Detect Genomic Aberrations in Chronic Lymphocytic.

Capture of CK+ and CK− complex aneuploid CTCs in breast, ovarian, or colorectal cancer. Capture of CK+ and CK− complex aneuploid CTCs in breast, ovarian,

Cyclin E1 Is Amplified and Overexpressed in Osteosarcoma

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

Successful Application of a Direct Detection Slide-Based Sequential Phenotype/Genotype Assay Using Archived Bone Marrow Smears and Paraffin Embedded.

Role of Chromosome 3q Amplification in Lung Cancer

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

Gayatry Mohapatra, Rebecca A. Betensky, Ezra R

A New Role for Hypoxia in Tumor Progression

Telomeric IGH Losses Detectable by Fluorescence in Situ Hybridization in Chronic Lymphocytic Leukemia Reflect Somatic VH Recombination Events Iwona Wlodarska,

Cytogenetics Part 2 Dr. Mohammed Hussein

Introduction & haematological malignancies

The MLPA assay and application to diagnosis of DGS

MET amplification is selected in KRAS wild-type colorectal cancer samples from patients who developed resistance to anti-EGFR treatment. MET amplification.

Oncogenic mutations Oncogenic mutations Examples of oncogene activating mutations are depicted. (A) Gene amplification leading to increased expression.

A, heatmap of copy number alterations determined by array CGH for a panel of 79 frozen NSCLC samples. A, heatmap of copy number alterations determined.

Presentation transcript:

F.I.S.H. in PATHOLOGY; Applications,Answers..and More Allan Kennedy Lead Biomedical Scientist (Molecular Pathology) Glasgow Royal Infirmary

Samples  Tissue Imprints  Blood  Formalin-fixed, paraffin embedded sections-usually 1µm thick!

TECHNIQUE  ACCESS THE TARGET  DENATURE  HYBRIDISE  COUNTERSTAIN  VIEW

F.I.S.H. WORK STATION

Probe use  Enumeration; Loss or Amplification  Breakapart; specific gene targeted  Dual Fusion; specific translocations detected

ENUMERATION PROBES e.g. Her2, CLL

Ideogram Her2 probe set

Normal Her2

AMPLIFIED HER2

Her2 ; Polysomy

Monosomy 17 with Her2 Amplification

Chronic Lymphocytic Leukemia (C.L.L.)

CLL PROBE SET

CLL P53 (17p13.1) Red ATM (11q22.3) Green;

CLL P53 (17p13.1) Red ATM (11q22.3) Green; Loss of ATM associated with progressive disease

CLL:Loss of p53 associated with progressive disease and Fludarabine resistance P53 (17p13.1)Red ATM (11q22.3) Green

Oligodendroglial Tumours Loss of Heterozygosity 1p/19q ?

1p36 Red 1q25 Green Loss of 1p36

19p13 green 19q13 red Loss of 19q

DUAL COLOUR DUAL FUSION PROBES e.g.(t11:14, t14:18)

Molecular Pathogenetic Mechanisms in Rhabdomyosarcoma and Ewing’s Sarcoma

t14:18 Negative Chr14 Green (IgH) Chr18 Red (bcl2) CEP18 Aqua

T14:18 Positive Chr14 Green (IgH) Chr18 Red (bcl2) CEP18 Aqua Translocations Yellow

BREAKAPART PROBES e.g. cMyc

cMyc Negative

C-Myc (8q24) Positive

ANSWERS

? Burkitt’s Lymphoma ?DLBCL LCA,CD10,CD20,bcl6 ;Positive TdT,CD5,MUM-1; Negative  FISH REQUEST FOR c-Myct8:14t14:18

C-Myc (8q24) Positive

t8:14 Positive

t14:18 Negative - Burkitt’s Lymphoma

? Burkitt ?Follicular Lymphoma  CD20,CD10,; Positive  CD5,Cyclin D1,CD23,bcl2; Negative

Myc negative

t14:18 (IgH / bcl 2) positive

Bcl6 (3q27) translocation negative; Follicular Lymphoma

Sarcomas

Molecular Diagnosis of Sarcomas TumorTranslocation Fusion Gene Ewing/PNETt(11;22)(q24;q12)EWS/FLI1 t(21;22)(q22;q12)EWS/ERG Alveolar Rhabdomyosarcoma t(2;13)(q35;q14)PAX3/FKHR t(1;13)(p36;q14)PAX7/FKHR Desmoplastic small round cell tumor t(11;22)(p13;q12)EWS/WT1 Synovial Sarcoma t(X;18)(p11.2;q11.2)SYT/SSX1+2 Congenital Fibrosarcoma t(12;15)(p13;q25)ETV6/NTRK3 Clear Cell Sarcoma t(12;22)(q13;q12)EWS/ATF1

Sarcoma  ? Ewings Sarcoma  ? Synovial sarcoma

Syt-Negative

EWS Positive-Ewings Sarcoma

? ALVEOLAR RHABDOMYOSARCOMA TEST FOR; t1:13t2:13 FKHR breakapart

t1:13 (Pax7 green/FKHR red ) Negative

t2:13 (Pax 3 green /FKHR red) Positive

FKHR (13q14) Positive

ANSWERS…and MORE

CLL P53 (17p13.1) Red ATM (11q22.3) Green;

t11:14 IgH 14q32 Green Bcl2 18q21 Red CEP11 aqua

Amplified Her2 Ratio 2.3

Amplified Her2 Ratio 10

Her2 ; Fusions?

Chromosome 17

Her2 gene

Her2 ; Triple Bandpass filter

Her2 gene

Chromosome 17

Her2 ; Homogeneously Stained Regions (HSR’s)

Her2;Double Minutes

 Need to record all observations-whether expected or not!  Need to collaborate across centres  Need close co-operation with Pathologists  Boundaries between Pathology, Genetics and Haematology rapidly blurring

Thank You