Philadelphia ChromosomeImportance in the oncologic diagnosis

Faculdade Medicina Universidade de CoimbraBiologia Celular e Molecular II2012-2013

Elaborated by: José Duarte Luís Oliveira Pedro PereiraTurma 9

INTRODUCTION

In site http://articles.philly.com/2010-09-29/news/24980608_1_chromosome-chronic-myeloid-leukemia-leukemia-cells

PHILADELPHIA CHROMOSOME (Ph)

Discovered in 1960 by Peter C. Nowel and David Hungerford;

First oncological gene documented;

Advances in cytogenetics.

ORIGIN

Results from a reciprocal translocation t(9;22) (q34;q11)

Defective chromosome 22 is designated Philadelphia Chromosome

Quimerical gene BCR-ABL is produced

In site http://www.medicinageriatrica.com.br/tag/cromossomo-filadelfia/

Chromosome 9

Chromosome 22

Normal Chromosomes Chromosomes Break Changed Chromosomes

Philadelphia Chromosome

Changed Chromosome 9

In site http://www.unesp.br/prope/projtecn/Saude/saude48a.htm

ORIGIN

FUSION GENE

Increases the tyrosine-kinase activity of ABL; Active; doesn’t need activation by other cellular messaging proteins; Activates some cell cycle-controlling proteins and enzymes, speeding up cell

division; Inhibits DNA repair; Increased proliferation in response to growth factors; Increased resistance to apoptosis, and alteration of their adhesion properties.

In site http://www.biologyreference.com/Oc-Ph/Oncogenes-and-Cancer-Cells.html

FUSION GENE

Three distinct forms:p190BCR/ABL

p210BCR/ABL

p230BCR/ABL

Leucemia

LEUKEMIA

Malignant tumor of bone marrow hematopoietic cells, that leads to the accumulation of the blast cells throughout the body.

In site http://www.cancer.gov/cancertopics/pdq/treatment/CML/Patient/page1

CANCERS RELATED TO PHILADELPHIA CHROMOSOME

The philadelphia chromosome is

present in 95% of people with CML

p210BCR/ABL

Acute Lymphoblastic

Leukemia (ALL) - 25–30% in adult

and 2–10% in pediatric cases

Chronic Myelogenous

Leukemia (CML)

Acute Lymphoblastic

Leukemia (ALL)

Acute Myelogenous

Leukemia (AML)

45–60% in adult and 80% in

pediatric casesp190BCR/ABL

Ph chromosome appears

occasionally;

most common in adults

p230BCR/ABL

Chronic phase (4-6 years):- Asymptomatic;- Thrombocytosis;- Leukocytosis;- Blast cell count less than 10%.

Accelerated phase (18 months): - Splenomegaly; - Thrombocytopenia; - Leukocytosis treatment resistant; - increased blast cells (10-30%) in peripheral blood and bone marrow; - Clonal evolution.

Blastic /acute phase (survival 2-4mounths) - ≥ 30% blast cells in bone marrow; - More resistant to treatment.

- Thrombocytopenia;- Leukocytopenia;- Vulnerability to bruising, bleeding and infections;- Lymphadenopathy;- Splenomegaly;- Pain in the bones or joints.

Chronic myelogenous leukemia Acute lymphoblastic leukemia Ph (+)

(Morales et al, 2010)

CLINICAL PRESENTATION

DIAGNOSIS

CLINICALLY ORIENTED:Splenomegaly

Hemogram

(blasts, basophiles, eosinophils)

Myelogram

Bone marrow biopsy

REFERRAL TO MOLECULAR AND CYTOGENETIC TESTS

In site http://iahealth.net/acute-myeloid-leukemia/

• PHILADELPHIA CHROMOSOME IDENTIFICATION:– Peripheral blood and bone marrow samples submitted to:

Southern Blotting Traditional PCR RT-PCR RQ-PCR (high sensitivity) Anchor-PET

Kariotype analysis (gold standard) FISH

CHROMOSOME ALTERATIONS

BCR/ABL transcripts

IDENTIFICATION and/or

QUANTIFICATION

VISUALIZATION TECHNIQUES

VISUALIZATION TECHNIQUES

Karyotype Analysis

In site http://www.citogene.com.br/Citogenetica-De-Cancer.aspx

VISUALIZATION TECHNIQUES

In site http://pt.wikipedia.org/wiki/Leucemia_mieloide_cr%C3%B4nica

Fluorescent hybridization in situ (FISH)

VISUALIZATION TECHNIQUES

In site http://www.scielo.br/scielo.php?pid=S0104-42301998000300015&script=sci_arttext

Polymerase Chain Reaction (PCR)

TREATMENT

CHEMOTHERAPY DONOR LYNPHOCYTE INFUSION TARGET THERAPY BIOLOGICAL THERAPY SURGERY HIGH DOSE CHEMOTHERAPY WITH

STEM CELLS TRANSPLANT

characteristic genetic abnormality of CML:

Philadelphia chromosome

protein BCR-ABL

tyrosine kinase

inhibitor of theBCR-ABL tyrosine

kinase

inhibitor of theBCR-ABL tyrosine

kinase

TARGET THERAPY

TARGET THERAPY

IMATINIB MESYLATE

Excelent therapeutic eficiency

Low toxicity

Blocks ATP binding site of BCR-ABL

Short time side effects

Resistance to imatinib

In site http://the-medical-dictionary.com/imatinib_mesylate.htm

PHILADELPHIA CHROMOSOME WAS THE ONSET OF TARGET THERAPY IN CANCER

TARGET THERAPY

SECOND GENERATION TKI

DASATINIB

NILOTINIB

BOSUTINIB

DCC-2036

AP24534

AT9283

REFERENCES

• Almeida A, Castro I, Coutinho J, Guerra L, Marques H, Pereira AM. Recomendações para o Diagnóstico, Tratamento e Monitorização da Leucemia Mielóide Crónica. Acta Med Port 2009; 22: 537-544• Cayuela J-M, Macintyre E, Darlington M, Abdelali RB, Fund X, Villarese P, Tulliez M, Raffoux E, Sigaux F, Réa D, and Seror V. Cartridge-based automated BCR-ABL1 mRNA quantification: solving the issues of standardization, at what cost?. Haematologica 2011;96(5):664-671.• Chauffaille ML. Análise citogenética e FISH no monitoramento da LMC em tratamento com inibidores da tirosino quinase . Rev. Bras. Hematol. Hemoter. 2008, 30 (Imagem FIsh)• Douglas, H; Robert A. W. – Hallmarks of Cancer: next generation. Elsevier. March 4, 2001. • Druker B. J. [et al]. Activity of a Specific Inhibitor of the BCR-ABL Tyrosine Kinase In the Blast Crisis of Chronic Myeloid Leukemia And Acute Lymphoblastic Leukemia With the Philadelphia Chromosome. N Engl J Med, Vol. 344, No. 14 April 5, 2001.• Funke, V. et al (2010). Leucemia Mielóide Crónica e outra doenças mieloprofilerativas crónicas. Revista Brasileira de Hematologia e Hemoterapia. 32 (Supl.1):71-90.• Goodsell D. S. The Molecular Perspective: c-Abl Tyrosine Kinase. The Oncologist 2005;10:758–759 • Grando AC; Wagner SC. Avaliação laboratorial da doença residual mínima na leucemia mielóide crônica por Real-Time PCR. J Bras Patol Med Lab. 2008, 44(6): 433-440• Griffiths, Anthony J. F. [et al]. Introdução à genética. 8ªedição. Rio de Janeiro: Guanabara-Koogan, 2006.• Guttmacher A. E., Collins F. S. Molecular Diagnosis of the Hematologic Cancers. N Engl J Med 2003;348:1777-85. •Hanahan D., Weinberg R. A. Hallmarks of Cancer: The Next Generation. Cell 144, March 4. Elsevier Inc, 2011. • Harrison – Medicina Interna. 17ª Edição, vol.I:683-686. •Hughes T, et al. Monitoring CML patients responding to treatment with tyrosine kinase inhibitors: review and recommendations for harmonizing current methodology for detecting BCR-ABL transcripts and kinase domain mutations and for expressing results. BLOOD. 2006, 108(1) 28-37• Knowles M., Selby P. Introduction to the Cellular and Molecular Biology of Cancer. 4 th ed. 2005, Oxford University Press. 189-193, 203-204, 219-226, 2005 • Koo, H. (2011). Philadelphia Chromossome-Positive Acute Lymphoblastic Leukemia in Childhood. Korean J Pediatr. 54 (3): 106-110.