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Hematolojik malignitelerde konvansiyonel sitogenetik, moleküler sitogenetik ve moleküler genetik sonuçlarının değerlendirilmesi

Year 2020, Volume: 27 Issue: 4, 547 - 554, 25.12.2020
https://doi.org/10.17343/sdutfd.813394

Abstract

Amaç: Hematolojik maligniteler, kemik iliği kaynaklı hücrelerin neoplazmalarıdır. Son çalışmalar, bu malignitelerin çoğunun sayısal ve yapısal kromozomal anormallikler içerdiğini göstermektedir. Spesifik gen düzeyindeki değişiklikler, bu hastalıkların tanı ve prognoz değerlendirmesinde çok önemlidir. Çalışmamız, bu genetik değişikliklerin bir kısmını ve bunların hematolojik malignitelerin etiyolojisi ve prognozu üzerindeki etkilerini araştırmayı amaçlamaktadır.
Gereç ve Yöntem: Bu çalışmaya Süleyman Demirel Üniversitesi Tıp Fakültesi Hematoloji Anabilim Dalı'na hematolojik malignite ön tanısı veya tanısı ile başvuran 110 hasta dahil edildi.
Akut Miyeloid Lösemi (AML), Kronik Miyeloid Lösemi (CML), Kronik Lenfoblastik Lösemi (CLL), Miyelodisplastik Sendrom (MDS), Kronik Miyeloproliferatif ( CMPD), Malign Plazma Hücreli Neoplazm (MPCN) ve Lenfoma hastalıklarına sahip yedi grubun kemik iliği kültürleri yapılarak üç farklı analiz yöntemi ile incelendi. Sayısal ve yapısal kromozomal değişiklikler sitogenetik kromozom analizi ve Floresan In Situ Hibridizasyon (FISH) yöntemleri ile incelendi ve JAK-2 V617F mutasyonu Real-Time PCR (RT-PCR) ile analiz edildi.
Bulgular: CML için t (9; 22) mutasyonu, AML için t (15; 17); MDS için del (5q) ve del (7q); lenfoma, CLL ve MPCN için del (13q14) ve del (17p13) mutasyonlarının etkili olduğu bulundu. Ayrıca RT-PCR sonuçlarına göre JAK-2 V617F mutasyonları, CMPD hastalığı alt gruplarında% 43'ten% 92'ye değişen oranlarda etkili bulunmuştur.
Sonuç: Elde ettiğimiz bulgular ışığında, hematolojik malignensili hastaların rutinde çalışılan mevcut genetik analizlerine ek olarak belirlediğimiz mutasyonlarında incelenmesinin hastalığın tanı ve prognozunun değerlendirilmesine katkıda bulunacağını düşünmekteyiz.

Supporting Institution

Süleyman Demirel Üniversitesi Bilimsel Araştırma Projeleri

Project Number

3460 / YL1–13

References

  • Heerema NA. Cytogenetic analysis of hematologic malignant diseases. The AGT Cytogenetics Laboratory Manual. 2017:499-575.
  • Dewald W G. Cytogenetic and FISH studies in myelodysplasia, acute myeloid leukemia, chronic lymphocytic leukemia and lymphoma. International journal of hematology. 2002;76:65-74.
  • Sultana TA, Mottalib MA, Islam MS, Khan MA, Choudhury S. rt-PCR method for diagnosis and follow-up of hematological malignancies: first approach in Bangladesh. Bangladesh Medical Research Council Bulletin. 2008;34(1):1-11.
  • Hokland P, Pallisgaard N, editors. Integration of molecular methods for detection of balanced translocations in the diagnosis and follow-up of patients with leukemia. Seminars in hematology; 2000: Elsevier.
  • Rowley J, Mitelman F. Principles of molecular cell biology of cancer: chromosome abnormalities in human cancer and leukemia. Cancer: Principles & Practice of Oncology DeVita, VT, Hellman, S, Rosenberg, SA,(Eds), Philadelphia: JB Lippincott Company. 1993:67-91.
  • Sreekantaiah C. FISH panels for hematologic malignancies. Cytogenetic and genome research. 2007;118(2-4):284-96. Dewald GW, Wyatt WA, Juneau AL, Carlson RO, Zinsmeister AR, Jalal SM, et al. Highly sensitive fluorescence in situ hybridization method to detect double BCR/ABL fusion and monitor response to therapy in chronic myeloid leukemia. Blood. 1998;91(9):3357-65.
  • Pallisgaard N, Hokland P, Riishøj DC, Pedersen B, Jørgensen P. Multiplex reverse transcription-polymerase chain reaction for simultaneous screening of 29 translocations and chromosomal aberrations in acute leukemia. Blood. 1998;92(2):574-88.
  • Hochhaus A, Weisser A, La Rosee P, Emig M, Müller M, Saussele S, et al. Detection and quantification of residual disease in chronic myelogenous leukemia. Leukemia. 2000;14(6):998.
  • Hermans A, Heisterkamp N, von Lindern M, van Baal S, Meijer D, van der Plas D, et al. Unique fusion of bcr and c-abl genes in Philadelphia chromosome positive acute lymphoblastic leukemia. Cell. 1987;51(1):33-40. Rabbitts T. Chromosomal translocations in human cancer. Nature. 1994;372(6502):143.
  • Willis T, Zalcberg I, Jadayel D, Coignet L, Stul M, Treleaven J, et al. Molecular cloning of translocation t (1; 14)(q21; q32) defines a novel gene (BCL9) at chromosome 1q21. Blood. 1997;90(10):1720-27.
  • Barnes DJ, Melo JV. Cytogenetic and molecular genetic aspects of chronic myeloid leukaemia. Acta haematologica. 2002;108(4):180-202.
  • Sazawal S, Bajaj J, Chikkara S, Jain S, Bhargava R, Mahapatra M, et al. Prevalence of JAK2 V617F mutation in Indian patients with chronic myeloproliferative disorders. Indian Journal of Medical Research. 2010;132(4):423.
  • Murugesan G, Aboudola S, Szpurka H, Verbic MA, Maciejewski JP, Tubbs RR, et al. Identification of the JAK2 V617F mutation in chronic myeloproliferative disorders using FRET probes and melting curve analysis. American Journal of Clinical Pathology. 2006;125(4):625-33.
  • Shin M-G, Kim HJ, Kim H-R, Lee S-Y, Lee I-K, Shin J-H, et al. Frequency of the JAK2 mutation and its usefulness as a marker for treatment response and disease progression in Korean patients with chronic myeloproliferative disorders. Am Soc Hematology; 2006.
  • Vardiman JW. The World Health Organization (WHO) classification of tumors of the hematopoietic and lymphoid tissues: an overview with emphasis on the myeloid neoplasms. Chemico-biological interactions. 2010;184(1-2):16-20. Başaran N. Tıbbi genetik: ders kitabı: Anadolu Üniversitesi; 1984.
  • Miranda RN, Mark H, Medeiros LJ. Fluorescent in situ hybridization in routinely processed bone marrow aspirate clot and core biopsy sections. The American journal of pathology. 1994;145(6):1309.
  • Ou J, Vergilio JA, Bagg A. Molecular diagnosis and monitoring in the clinical management of patients with chronic myelogenous leukemia treated with tyrosine kinase inhibitors. American journal of hematology. 2008;83(4):296-302.
  • Kantarjian H, Schiffer C, Jones D, Cortes J. Monitoring the response and course of chronic myeloid leukemia in the modern era of BCR-ABL tyrosine kinase inhibitors: practical advice on the use and interpretation of monitoring methods. Blood, The Journal of the American Society of Hematology. 2008;111(4):1774-80.
  • Bain BJ. Overview. Best Practice & Research Clinical Haematology. 2001;14(3):463-77.
  • Linenberger ML, Hong T, Flowers D, Sievers EL, Gooley TA, Bennett JM, et al. Multidrug-resistance phenotype and clinical responses to gemtuzumab ozogamicin. Blood. 2001;98(4):988-94.
  • Mitelman F, Heim S. Quantitative acute leukemia cytogenetics. Genes, Chromosomes and Cancer. 1992;5(1):57-66. Downing JR. The AML1‐ETO chimaeric transcription factor in acute myeloid leukaemia: biology and clinical significance. British journal of haematology. 1999;106(2):296-308.
  • Foon KA, Rai KR, Gale RP. Chronic lymphocytic leukemia: new insights into biology and therapy. Annals of Internal Medicine. 1990;113(7):525-39.
  • Choi W, Kim M, Lim J, Han K, Lee S, Lee JW, et al. Four cases of chronic myelogenous leukemia in mixed phenotype blast phase at initial presentation mimicking mixed phenotype acute leukemia with t (9; 22). Annals of laboratory medicine. 2014;34(1):60-3.
  • Maciejewski JP, Selleri C. Evolution of clonal cytogenetic abnormalities in aplastic anemia. Leukemia & lymphoma. 2004;45(3):433-40.
  • Komrokji RS, Bennett JM. What Is “WHO “? Myelodysplastic Syndromes Classification. Clinical Leukemia. 2008;2(1):20-7. Lakatošová M, Holečková B. Fluorescence in situ hybridisation. Biologia. 2007;62(3):243-50.
  • Tefferi A, Vardiman J. Classification and diagnosis of myeloproliferative neoplasms: the 2008 World Health Organization criteria and point-of-care diagnostic algorithms. Leukemia. 2008;22(1):14.
  • Levine RL, Pardanani A, Tefferi A, Gilliland DG. Role of JAK2 in the pathogenesis and therapy of myeloproliferative disorders. Nature reviews cancer. 2007;7(9):673.
  • James C, Ugo V, Le Couédic J-P, Staerk J, Delhommeau F, Lacout C, et al. A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature. 2005;434(7037):1144.
  • Hussein K, Bock O, Seegers A, Flasshove M, Henneke F, Buesche G, et al. Myelofibrosis evolving during imatinib treatment of a chronic myeloproliferative disease with coexisting BCR-ABL translocation and JAK2V617F mutation. Blood. 2007;109(9):4106-7.
  • Karkucak M, Yakut T, Ozkocaman V, Ozkalemkas F, Ali R, Bayram M, et al. Evaluation of the JAK2-V617F gene mutation in Turkish patients with essential thrombocythemia and polycythemia vera. Molecular biology reports. 2012;39(9):8663-7.
  • Çetinkaya S. Trakya Üniversitesi Tıp Fakültesi Hematoloji Bilim Dalına başvuran hastalarda JAK 2 geni nokta mutasyonu ve hastalık ilişkisinin değerlendirilerek fenotip-genotip ilişkisinin kurulması: Trakya Üniversitesi Sağlık Bilimleri Enstitüsü; 2012.

Evaluation of conventional cytogenetic, molecular cytogenetics and molecular genetics results in hematological malignances

Year 2020, Volume: 27 Issue: 4, 547 - 554, 25.12.2020
https://doi.org/10.17343/sdutfd.813394

Abstract

Objective: Hematological malignancies are neoplasms of bone marrow-derived cells. Recent studies show that most of these malignancies contain numerical and structural chromosomal abnormalities. These specific gene-level changes are crucial at diagnosis and prognosis evaluation of these diseases. Our study aims to investigate some of these genetic changes and their effect on the etiology and prognosis of hematological malignancies.
Materials and Methods: In this study, 110 patients who were admitted to the Department of Hematology of Süleyman Demirel University Faculty of Medicine with a pre-diagnosis or diagnosis of Hematologic Malignancy were included. Three different analyses applied to the cultured bone marrow tissue samples of seven groups of hematology patients who suffered from Acute Myeloid Leukemia (AML), Chronic Myeloid Leukemia (CML), Chronic Lymphoblastic Leukemia (CLL), Myelodysplastic Syndrome (MDS), Chronic Myeloproliferative Disease (CMPD), Malignant Plasma Cell Neoplasm (MPCN) and Lymphoma diseases.Numerical and structural chromosomal changes were examined by cytogenetic chromosome analysis and Fluorescent In Situ Hybridization (FISH) methods and JAK-2 V617F mutation was analyzed by Real-Time PCR (RT-PCR).
Results: Some mutations were found important in particular diseases such as t(9;22) mutation for CML, t(15;17) for AML; del(5q) and del(7q) for MDS; del(13q14) and del(17p13) mutations for lymphoma, CLL and MPCN. JAK-2 V617F mutations were found effective on CMPD disease subgroups at rates changing from 43% up to 92% according to the RT-PCR results.
Conclusion: As a result of our findings, we think that examining patients with hematologic malignancies in their mutations, in addition to the routinely studied existing genetic analyzes, will contribute to the evaluation of the diagnosis and prognosis of the disease.

Project Number

3460 / YL1–13

References

  • Heerema NA. Cytogenetic analysis of hematologic malignant diseases. The AGT Cytogenetics Laboratory Manual. 2017:499-575.
  • Dewald W G. Cytogenetic and FISH studies in myelodysplasia, acute myeloid leukemia, chronic lymphocytic leukemia and lymphoma. International journal of hematology. 2002;76:65-74.
  • Sultana TA, Mottalib MA, Islam MS, Khan MA, Choudhury S. rt-PCR method for diagnosis and follow-up of hematological malignancies: first approach in Bangladesh. Bangladesh Medical Research Council Bulletin. 2008;34(1):1-11.
  • Hokland P, Pallisgaard N, editors. Integration of molecular methods for detection of balanced translocations in the diagnosis and follow-up of patients with leukemia. Seminars in hematology; 2000: Elsevier.
  • Rowley J, Mitelman F. Principles of molecular cell biology of cancer: chromosome abnormalities in human cancer and leukemia. Cancer: Principles & Practice of Oncology DeVita, VT, Hellman, S, Rosenberg, SA,(Eds), Philadelphia: JB Lippincott Company. 1993:67-91.
  • Sreekantaiah C. FISH panels for hematologic malignancies. Cytogenetic and genome research. 2007;118(2-4):284-96. Dewald GW, Wyatt WA, Juneau AL, Carlson RO, Zinsmeister AR, Jalal SM, et al. Highly sensitive fluorescence in situ hybridization method to detect double BCR/ABL fusion and monitor response to therapy in chronic myeloid leukemia. Blood. 1998;91(9):3357-65.
  • Pallisgaard N, Hokland P, Riishøj DC, Pedersen B, Jørgensen P. Multiplex reverse transcription-polymerase chain reaction for simultaneous screening of 29 translocations and chromosomal aberrations in acute leukemia. Blood. 1998;92(2):574-88.
  • Hochhaus A, Weisser A, La Rosee P, Emig M, Müller M, Saussele S, et al. Detection and quantification of residual disease in chronic myelogenous leukemia. Leukemia. 2000;14(6):998.
  • Hermans A, Heisterkamp N, von Lindern M, van Baal S, Meijer D, van der Plas D, et al. Unique fusion of bcr and c-abl genes in Philadelphia chromosome positive acute lymphoblastic leukemia. Cell. 1987;51(1):33-40. Rabbitts T. Chromosomal translocations in human cancer. Nature. 1994;372(6502):143.
  • Willis T, Zalcberg I, Jadayel D, Coignet L, Stul M, Treleaven J, et al. Molecular cloning of translocation t (1; 14)(q21; q32) defines a novel gene (BCL9) at chromosome 1q21. Blood. 1997;90(10):1720-27.
  • Barnes DJ, Melo JV. Cytogenetic and molecular genetic aspects of chronic myeloid leukaemia. Acta haematologica. 2002;108(4):180-202.
  • Sazawal S, Bajaj J, Chikkara S, Jain S, Bhargava R, Mahapatra M, et al. Prevalence of JAK2 V617F mutation in Indian patients with chronic myeloproliferative disorders. Indian Journal of Medical Research. 2010;132(4):423.
  • Murugesan G, Aboudola S, Szpurka H, Verbic MA, Maciejewski JP, Tubbs RR, et al. Identification of the JAK2 V617F mutation in chronic myeloproliferative disorders using FRET probes and melting curve analysis. American Journal of Clinical Pathology. 2006;125(4):625-33.
  • Shin M-G, Kim HJ, Kim H-R, Lee S-Y, Lee I-K, Shin J-H, et al. Frequency of the JAK2 mutation and its usefulness as a marker for treatment response and disease progression in Korean patients with chronic myeloproliferative disorders. Am Soc Hematology; 2006.
  • Vardiman JW. The World Health Organization (WHO) classification of tumors of the hematopoietic and lymphoid tissues: an overview with emphasis on the myeloid neoplasms. Chemico-biological interactions. 2010;184(1-2):16-20. Başaran N. Tıbbi genetik: ders kitabı: Anadolu Üniversitesi; 1984.
  • Miranda RN, Mark H, Medeiros LJ. Fluorescent in situ hybridization in routinely processed bone marrow aspirate clot and core biopsy sections. The American journal of pathology. 1994;145(6):1309.
  • Ou J, Vergilio JA, Bagg A. Molecular diagnosis and monitoring in the clinical management of patients with chronic myelogenous leukemia treated with tyrosine kinase inhibitors. American journal of hematology. 2008;83(4):296-302.
  • Kantarjian H, Schiffer C, Jones D, Cortes J. Monitoring the response and course of chronic myeloid leukemia in the modern era of BCR-ABL tyrosine kinase inhibitors: practical advice on the use and interpretation of monitoring methods. Blood, The Journal of the American Society of Hematology. 2008;111(4):1774-80.
  • Bain BJ. Overview. Best Practice & Research Clinical Haematology. 2001;14(3):463-77.
  • Linenberger ML, Hong T, Flowers D, Sievers EL, Gooley TA, Bennett JM, et al. Multidrug-resistance phenotype and clinical responses to gemtuzumab ozogamicin. Blood. 2001;98(4):988-94.
  • Mitelman F, Heim S. Quantitative acute leukemia cytogenetics. Genes, Chromosomes and Cancer. 1992;5(1):57-66. Downing JR. The AML1‐ETO chimaeric transcription factor in acute myeloid leukaemia: biology and clinical significance. British journal of haematology. 1999;106(2):296-308.
  • Foon KA, Rai KR, Gale RP. Chronic lymphocytic leukemia: new insights into biology and therapy. Annals of Internal Medicine. 1990;113(7):525-39.
  • Choi W, Kim M, Lim J, Han K, Lee S, Lee JW, et al. Four cases of chronic myelogenous leukemia in mixed phenotype blast phase at initial presentation mimicking mixed phenotype acute leukemia with t (9; 22). Annals of laboratory medicine. 2014;34(1):60-3.
  • Maciejewski JP, Selleri C. Evolution of clonal cytogenetic abnormalities in aplastic anemia. Leukemia & lymphoma. 2004;45(3):433-40.
  • Komrokji RS, Bennett JM. What Is “WHO “? Myelodysplastic Syndromes Classification. Clinical Leukemia. 2008;2(1):20-7. Lakatošová M, Holečková B. Fluorescence in situ hybridisation. Biologia. 2007;62(3):243-50.
  • Tefferi A, Vardiman J. Classification and diagnosis of myeloproliferative neoplasms: the 2008 World Health Organization criteria and point-of-care diagnostic algorithms. Leukemia. 2008;22(1):14.
  • Levine RL, Pardanani A, Tefferi A, Gilliland DG. Role of JAK2 in the pathogenesis and therapy of myeloproliferative disorders. Nature reviews cancer. 2007;7(9):673.
  • James C, Ugo V, Le Couédic J-P, Staerk J, Delhommeau F, Lacout C, et al. A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature. 2005;434(7037):1144.
  • Hussein K, Bock O, Seegers A, Flasshove M, Henneke F, Buesche G, et al. Myelofibrosis evolving during imatinib treatment of a chronic myeloproliferative disease with coexisting BCR-ABL translocation and JAK2V617F mutation. Blood. 2007;109(9):4106-7.
  • Karkucak M, Yakut T, Ozkocaman V, Ozkalemkas F, Ali R, Bayram M, et al. Evaluation of the JAK2-V617F gene mutation in Turkish patients with essential thrombocythemia and polycythemia vera. Molecular biology reports. 2012;39(9):8663-7.
  • Çetinkaya S. Trakya Üniversitesi Tıp Fakültesi Hematoloji Bilim Dalına başvuran hastalarda JAK 2 geni nokta mutasyonu ve hastalık ilişkisinin değerlendirilerek fenotip-genotip ilişkisinin kurulması: Trakya Üniversitesi Sağlık Bilimleri Enstitüsü; 2012.
There are 31 citations in total.

Details

Primary Language English
Subjects Clinical Sciences
Journal Section Araştırma Makaleleri
Authors

Pınar Koşar 0000-0003-2602-5145

Muhammet Yusuf Tepebaşı 0000-0002-1087-4874

Emine Güçhan Alanoğlu

Project Number 3460 / YL1–13
Publication Date December 25, 2020
Submission Date October 20, 2020
Acceptance Date December 9, 2020
Published in Issue Year 2020 Volume: 27 Issue: 4

Cite

Vancouver Koşar P, Tepebaşı MY, Alanoğlu EG. Evaluation of conventional cytogenetic, molecular cytogenetics and molecular genetics results in hematological malignances. Med J SDU. 2020;27(4):547-54.

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