Cancer Cytogenetic Laboratory

Chromosomal aberrations associated with malignant tumors are detected by conventional and molecular cytogenetic techniques in our laboratory. The majority of received samples, including bone marrow aspirates, peripheral blood samples, lymph nodes, CSF, effusions and frozen or FFPE tumor tissues, taken from patients suffering from oncological and hematological diseases are investigated. Karyotyping following G-banding is capable of characterizing the whole chromosome set of dividing cells identifying numerical abnormalities and structural aberrations affecting at least 5-10 Mbs. Cryptic aberrations undetectable by conventional cytogenetics are screened by target-specific fluorescence in situ hybridization, a technique that may be applied in case of any cytological and histological samples without the need for cell cultures. At present, FISH is used for the detection of diagnostically important or therapy relevant aberrations in oncological cases. FISH labeling is also performed on metaphase preparations if the evaluation of karyotypes is difficult because of its complexity. Our cytogenetic core facility covers not only the Medical School and Clinical Center of the University of Pécs but also larger regions involving mostly the South-West of Hungary.

Research

Interphase-FISH is a robust method of oncopathological diagnostics. Preparations are evaluated manually in most laboratories, however, some non-negligible problems arise from this approach: (i) a huge number of nuclei need to be evaluated to ensure statistical reliability when the ratio of positivity is very low, and this is time-consuming; (ii) bias of investigators may cause over-, or underestimation of positive events, especially at high or low concentrations of positivity, respectively; (iii) both interobserver and interlaboratory variability may be significant; (iv) an objective definition of translocation-related fusion/colocalization of signals is lacking, despite the crucial importance of this parameter. These difficulties can be eliminated by using motorized microscopy and digital image analysis.

We have introduced the following automated, 3D signal pattern evaluating techniques in the last few years: (i) genotyping on cytological preparations (detection of BCR/ABL juxtaposition in chronic myeloid leukemia); (ii) combined fluorescent immunophenotyping and genotyping on cytological samples for the follow-up of minimal residual disease (identification of leukemic cells by consecutive CD10 immunolabeling followed by FISH analysis of ETV6/RUNX1 fusion in pediatric acute lymphoblastic leukemia); (iii) combined chromogenic immunophenotyping and molecular cytogenetic analysis using four different genetic markers on cytological samples (CK7 immunolabeling followed by UroVysion analysis in bladder cancer); (iv) genotyping on histological samples (investigation of IGH/CCND1 and IGH/BCL-2 translocations in mantle cell lymphoma and follicular lymphoma, respectively). We think these standardized techniques contribute to the development of pathological diagnostics, therefore we would like to enlarge our automated interphase cytogenetic potential and capacity in the future.

 

 

Publications

T. Tornóczky, E. Kálmán, Z. Sápi, Z. Orosz, L. Pajor: Cytogenetic abnormalities of alveolar soft-part sarcomas using interphase fluorescent in situ hybridization: trisomy for chromosome 7 and monosomy for chromosomes 8 and 18 seem to be characteristic of the tumor. Virchows Arch. 2001 Feb;438(2):173-80.

Tamás Tornóczky, Endre Kálmán, Pál G Kajtár, Tibor Nyári, Andrew D J Pearson, Deborah A Tweddle, Julian Board, Hiroyuki Shimada: Large cell neuroblastoma: a distinct phenotype of neuroblastoma with aggressive clinical behavior. Cancer. 2004 Jan 15;100(2):390-7.

Kajtár B, Méhes G, Lörch T, Deák L, Kneifné M, Alpár D, and Pajor L: Automated Fluorescent In Situ Hybridization (FISH) Analysis of t(9;22)(q34;q11) in Interphase Nuclei. Cytometry Part A, 2006 Jun;69(6):506-14.

Alpár D, Kajtár B, Kneif M, Jáksó P, László R, Kereskai L, Pajor L: Automated detection of residual leukemic cells by consecutive immunolabeling for CD10 and FISH for ETV6/RUNX1 rearrangement in childhood acute lymphoblastic leukemia. Cancer Genetics and Cytogenetics, 2007 Feb;173(1):23-30.

Pajor G, Süle N, Alpár D, Kajtár B, Kneif M, Bollmann D, Somogyi L, Pajor L: Increased efficiency of detecting genetically aberrant cells by UroVysion test on voided urine specimens using automated immunophenotypical pre-selection of uroepithelial cells. Cytometry Part A, 2008 Mar;73A(3):259-65.

Alpár D, Hermesz J, László R, Kereskai L, Jáksó P, Pajor G, Pajor L, Kajtár B: Automated FISH analysis using dual-fusion and break-apart probes on paraffin-embedded tissue sections. Cytometry Part A, 2008 Jul;73(A):651-57.

László R, Alpár D, Kajtár B, Lacza Á, Ottóffy G, Kiss Cs, Bartyik K, Nagy K, Pajor L. Detection of early precursors of t(12;21) positive pediatric acute lymphoblastic leukemia during follow-up. Pediatric Blood and Cancer, 2010 Jan;54(1):158-60.

Alpár D, Nagy G, Hohoff C, Kajtár B, Bartyik K, Hermesz J, Jáksó P, Andrikovics H, Kereskai L, Pajor L. Sex chromosome changes after sex-mismatched allogeneic bone marrow transplantation can mislead the chimerism analysis. Pediatric Blood and Cancer, 2010 Dec;55:1239-42. 

Alpár D, Kereskai L. Three-step cytogenetic evolution in paediatric acute lymphoblastic leukaemia with t(12;21). BloodMed, 2010 Dec; 251.

Alpár D. Recurrent disease or donor cell leukemia? Brain teaser after allogeneic bone marrow transplantation. Chimerism, 2011 Jan;2(1):19-20.

Nagy Z, Kajtár B, Jáksó P, Dávid M, Kosztolányi S, Hermesz J, Kereskai L, Pajor L, Alpár D. Evolutionary sequence of cytogenetic aberrations during the oncogenesis of plasma cell disorders. Direct evidence at single cell level. Leukemia Research, 2011 Aug;35(8):1114-6.

Pajor G, Somogyi L, Melegh B, Alpár D, Kajtár B, Farkas L, Kneif M, Bollmann D, Pajor L, Süle N. Urovysion: considerations on modifying current evaluation scheme, including immunophenotypic targeting and locally set statistically derived diagnostic criteria. Cytometry Part A, 2011 May;79(5):375-82.

Tamás Tornóczky, Barna Bogner, Thomas Krausz, Gábor Ottóffy, Károly Szuhai: Angiomatoid fibrous histiocytoma: pleomorphic variant associated with multiplication of EWSR1-CREB1 fusion gene. Pathol Oncol Res. 2012 Apr;18(2):545-8.

Pajor G, Alpár D, Kajtár B, Melegh B, Somogyi L, Kneif M, Bollmann D, Süle N, Pajor L. Automated signal pattern evaluation of a bladder cancer specific multiprobe-FISH assay applying a user-trainable workstation. Microscopy Research and Technique, 2012 Jun;75(6):814-20.

Pajor G, Kajtár B, Pajor L, Alpár D. State-of-the-art FISHing: automated analysis of cytogenetic aberrations in interphase nuclei. Review. Cytometry A. 2012 Aug;81(8):649-63.

Alpár D, de Jong D, Savola S, Yigittop H, Kajtár B, Kereskai L, Pajor L, Szuhai K. MLPA is a powerful tool for detecting lymphoblastic transformation in chronic myeloid leukemia and revealing the clonal origin of relapse in pediatric acute lymphoblastic leukemia. Cancer Genet. 2012 Sep;205(9):465-9.

Haltrich I, Csóka M, Kovács G, Török D, Alpár D, Ottoffy G, Fekete G. Six cases of rare gene amplifications and multiple copy of fusion gene in childhood acute lymphoblastic leukemia. Pathol Oncol Res. 2013 Jan;19(1):123-8.

Alpar D, de Jong D, Holczer-Nagy Z, Kajtar B, Savola S, Jakso P, David M, Kosztolanyi S, Kereskai L, Pajor L, Szuhai K. Multiplex ligation-dependent probe amplification and fluorescence in situ hybridization are complementary techniques to detect cytogenetic abnormalities in multiple myeloma. Genes Chromosomes Cancer. 2013 Sep;52(9):785-93.

Kajtár B, Tornóczky T, Kálmán E, Kuzsner J, Hogendoorn PC, Szuhai K. CD99-positive undifferentiated round cell sarcoma diagnosed on fine needle aspiration cytology, later found to harbour a CIC-DUX4 translocation: a recently described entity. Cytopathology. 2014 Apr;25(2):129-32.

Semjen D, Kalman E, Tornoczky T, Szuhai K.: Further evidence of the existence of benign teratomas of the postpubertal testis. Am J Surg Pathol. 2014 Apr;38(4):580-1.

Alpár D, Pajor G, Varga P, Kajtár B, Pótó L, Mátics R, Vojcek A, Ottoffy G, Szuhai K, Pajor L. Sequential and hierarchical chromosomal changes and chromosome instability are distinct features of high hyperdiploid pediatric acute lymphoblastic leukemia. Pediatr Blood Cancer. 2014 Dec;61(12):2208-14.

Smuk G, Tornóczky T, Pajor L, Chudoba I, Kajtár B, Sárosi V, Pajor G. Immense random colocalization, revealed by automated high content image cytometry, seriously questions FISH as gold standard for detecting EML4-ALK fusion. Cytometry A. 2018 Jun;93(6):653-661.

Kosztolányi S, Kiss R, Atanesyan L, Gángó A, de Groot K, Steenkamer M, Jáksó P, Matolcsy A, Kajtár B, Pajor L, Szuhai K, Savola S, Bödör C, Alpár D. High-Throughput Copy Number Profiling by Digital Multiplex Ligation-Dependent Probe Amplification in Multiple Myeloma. J Mol Diagn. 2018 Nov;20(6):777-788.

Kosztolányi S, Horváth B, Hosnyánszki D, Kereskai L, Sziládi E, Jáksó P, Alizadeh H, Szuhai K, Alpár D, Kajtár B. [Molecular cytogenetic analyses of patients with plasma cell myeloma in Tolna and Baranya counties in Hungary]. Orv Hetil. 2019 Jun;160(24):944-951.

Gábor Smuk, Gábor Pajor, Károly Szuhai, Hans Morreau, Ildikó Kocsmár, Éva Kocsmár, László Pajor, Béla Kajtár, Veronika Sárosi, Gábor Lotz, Tamás Tornóczky: Attenuated isolated 3' signal: A highly challenging therapy relevant ALK FISH pattern in NSCLC. Lung Cancer. 2020 May;143:80-85.

Dávid Semjén, Krisztina Bíró, Emese Kapitány, Endre Kálmán, Tamás Tornóczky, Béla Kajtár: Histology, 12p status, and IMP3 expression separate subtypes in testicular teratomas. Virchows Arch. 2020 Jul;477(1):103-110.

 

Education

Our laboratory is involved in the Ph.D. course titled ’Molecular pathology in modern medicine: diagnostic, predictive and therapy-associated tests’, as well as training of laboratory analysts and laboratory researchers.

Team

Béla Kajtár, MD. Ph.D.

senior lecturer
+36-72-536-000/31843

e-mail: kajtar.bela@pte.hu

Bálint Horváth
biologist
+36/72-536-001/31505

Diána Hosnyánszki
biologist
+36/72-536-001/31505

Viktória Takács
biologist
+36/72-536-001/31505

Ivett Sepsei, BSc
medical diagnostic analyst
+36-72-536-000/31505

 

Links

http://atlasgeneticsoncology.org/
http://cgap.nci.nih.gov/Chromosomes/Mitelman