Využití interfázní fluorescenční in situ hybridizace pro analýzu CD34+ buněk v periferní krvi u nemocných s myelodysplastickými syndromy
Authors:
Š. Ransdorfová 1; J. Březinová 1; I. Šárová 1,2; M. Beličková 1; A. Jonášová 3; R. Neuwirtová 3; Z. Zemanová 2; J. Čermák 1; K. Michalová 1,2
Authors‘ workplace:
Ústav hematologie a krevní transfuze, Praha
1; Centrum nádorové cytogenetiky, Ústav lékařské biochemie a laboratorní diagnostiky VFN a 1. LF UK, Praha
2; 1. interní klinika VFN a 1. LF UK, Praha
3
Published in:
Transfuze Hematol. dnes,22, 2016, No. 2, p. 90-96.
Category:
Comprehensive Reports, Original Papers, Case Reports
Overview
Classical cytogenetic analysis of bone marrow cells is the standard for evaluating chromosomal aberrations in patients with myelodysplastic syndromes (MDS). However, there are several factors affecting the success of this method, such as the degree of cell proliferation, quality of preparations or size of pathological clone. In these cases, it is possible to use interphase fluorescence in situ hybridization (I-FISH), which allows detection of chromosomal aberrations in the nuclei of interphase cells. In this study, we used the I-FISH method to screen chromosomal aberrations in peripheral blood circulating CD34+ cells. Our objective was to evaluate the frequency of chromosomal aberrations in CD34+ cells isolated from peripheral blood, to compare this with the results of I-FISH analysis of bone marrow cells and to assess the significance of this method for monitoring the effect of therapy in patients with MDS.
We examined 22 samples of CD34+ cells separated from the peripheral blood of 16 patients with MDS (13 females, 3 males; 11 x 5q- syndromes, 4x MDS RCMD and 1x MDS/MPS). Four patients were investigated repeatedly.
I-FISH on the separated CD34+ cells confirmed the prior findings in bone marrow in all the patients examined – 14 patients had deletion of chromosome 5 in the 5q31 region, one patient had deletion of chromosome 20q11q12 and one patient had monosomy of chromosome 7. In our work, we demonstrated that isolation of CD34+ cells followed by I-FISH analysis is a practical, non-invasive method that can detect specific chromosomal abnormalities present in the pathological clone. This method has a predictive value and can be used for continuous disease control as well as for monitoring the pathological clone dynamics following treatment.
Key words:
CD34+ cells – I-FISH – myelodysplastic syndromes
Sources
1. Silverman LR, Demakos EP, Peterson BL, et al. Randomized controlled trial of azacitidine in patients with the myelodysplastic syndrome: a study of the cancer and leukemia group B. J Clin Oncol 2002; 20: 2429–2440.
2. Parker JE, Mufti GJ. Ineffective haemopoiesis and apoptosis in myelodysplastic syndromes. Br J Haematol 1998; 101: 220–230.
3. Germing U, Gattermann N, Strupp C, et al. Validation of WHO proposals for a new classification of primary myelodysplastic syndromes: a retrospective analysis of 1600 patients. Leukemia Res 2000; 24: 983–992.
4. Schanz J, Tüchler H, Solé F, et al. A new comprehensive cytogenetic scoring system for primary myelodysplastic syndromes (MDS) and oligoblastic AML after MDS derived from an international diabase merge. J Clin Oncol 2012; 30: 820–829.
5. Greenberg P, Tuechler H, Schanz J, et al. Revised international scoring system for myelodysplastic syndromes. Blood 2012; 120: 2454–2465.
6. Schanz J, Steidl C, Fonatsch C, et al. Coalesced multicenter analysis of 2,351 patients with myelodysplastic syndromes indicates an underestimation of poor-risk cytogenetics of myelodysplastic syndromes in the international prognostic scoring system. J Clin Oncol 2011; 29: 1963–1970.
7. Solé F, Espinet B, Sanz GF, et al. Incidence, characterization and prognostic significance of chromosomal abnormalities in 640 patients with primary myelodysplastic syndromes: Grupo Cooperativo Espanol de Citogenetica Hematologica. Br J Haematol 2000; 108: 346–356.
8. Vardiman JW, Thiele J, Arber DA. The 2008 revision of the World Health Organization (WHO) classification of myeloid neoplasms and acute leukemia: rationale and important changes. Blood 2009; 114: 937–951.
9. Nimer SD. Clinical management of myelodysplastic syndromes with interstitial deletion of chromosome 5q. J Clin Oncol 2006; 24: 2576–2582.
10. Van den Berghe H, Cassimann JJ, David G, et al. Distinct haematological disorder with deletion of long arm of no.5 chromosome. Nature 1974; 251: 437–438.
11. Jerez A, Gondek LP, Jankowska AM, et al. Topography, clinical and genomic correlates of 5q myeloid malignancies revisited. J Clinic Oncol 2012; 30: 1343–1349.
12. Neuwirtová R, Zemanová Z, Březinová J, et al. Jsme oprávněni řadit nemocné s dvěma samostatnými buněčnými klony a to s delecí 5q a s trizomií 8 jako podskupinu myelodysplatického syndromu typu 5q- syndromu? Transfuze Hematol Dnes 2014; 20: 25–31.
13. Vehmeyer K, Haase D, Alves F. Increased peripheral stem cell pool in MDS: an indication of disease progression? Leukemia Res 2001; 25: 955–959.
14. Braulke F, Schanz J, Jung K, et al. FISH analysis of circulating CD34+ cells as a new tool for genetic monitoring in MDS: Verification of the method and application to 27 MDS patients. Leukemia Res 2010; 34: 1296–1301.
15. Haase D, Feuring-Buske M, Schäfer C, et al. Cytogenetic analysis of CD34+ subpopulations in AML and MDS characterized by the expression of CD38 and CD 117. Leukemia 1997; 11: 674–679.
16. Shaffer LG, McGowan-Jordan J, Schmid M (eds). ISCN 2013: An international system for human cytogenetic nomenclature. S. Karger, Basel 2013.
17. Haase D, Fontasch C, Freund M, et al. Cytogenetic findings in 179 patients with myelodysplastic syndromes. Ann Hematol 1995; 70: 171–187.
18. Miltenyi S, Műller W, Weichel W, et al. High gradient magnetic cell separation with MACS. Cytometry 1990; 11: 231–238.
19. Lübbert M, Wijermans P, Kunzmann R, et al. Cytogenetic responses in high-risk myelodysplastic syndrome following low-dose treatment with the DNA methylation inhibitor 5-aza-2’-deoxycytidine. Br J Haematol 2001; 114: 349–357.
20. Braulke F, Jung K, Schanz J, et al. Molecular cytogenetic monitoring from CD34+ peripheral blood cells in myelodysplastic syndromes: first results from a prospective multicenter German diagnostic study. Leukemia Res 2013; 37: 900–906.
21. Braulke F, Platzbecker U, Müller-Thomas C, et al. Validation of cytogenetic risk groups according to International Prognostic Scoring Systems by peripheral blood CD34+FISH: results from a German diagnostic study in comparison with an international control group. Haematologica 2015; 100: 205–213.
Labels
Haematology Internal medicine Clinical oncologyArticle was published in
Transfusion and Haematology Today
2016 Issue 2
Most read in this issue
- Indications for allogeneic and autologous haematopoietic cell transplantations in the Czech Republic in 2016: Recommendations of the Transplant Section of the Czech Society of Haematology and the Czech Society of Oncology of the J. E. Purkyně Medical Association
- Single center experience with the treatment of older patients with myelodysplastic syndrome, not eligible for intensive therapy and its financial consequences
- Recommendations for the diagnosis and treatment of chronic lymphocytic leukaemia (CLL) Czech Group for Chronic Lymphocytic Leukaemia, section of the Czech Society of Haematology, CzMA
- Prophylaxis versus on demand treatment in adult patients with severe haemophilia A – FN Brno experience