Kronik Faz Kronik Miyeloid Lösemi Hastalarının Akış Sitometrik Analizi

Öz

Morfoloji ve sitogenetik için kemik iliği aspiratı ve periferik kan hücrelerinde kalitatif revers transkriptaz polimeraz zincir reaksiyonu, kronik miyeloid lösemi (KML) tanısı için zorunludur. Seçilmiş hastalarda kemik iliği biyopsisi ve floresan in situ hibridizasyon gerekli olabilir. Diğer testlerin ve teşhis prosedürlerinin uygulanması, bireysel hastanın özelliklerine bağlıdır. Akış sitometrisi birçok hematolojik malignitenin tanısında ve izlenmesinde önemli bir araç olmasına rağmen, KML'de sınırlı bir rolü vardır. Bu çalışmada KML hastalarımızın tanı anında ve takip sırasındaki CD45 dağılım sonuçlarını değerlendirdik. İmatinib ile tedavi edilen kronik fazdaki toplam 56 KML hastası (22 kadın ve 34 erkek) çalışmaya dahil edildi. Hastalar ortalama 8 (3-19) aylık takipliydi. Hastaların tanı ve takipteki tam kan sayımı parametreleri ile CD45/SSC sonuçları retrospektif olarak değerlendirildi. İki grup arasındaki ortalamaları karşılaştırmak için Wilcoxon T testi kullanıldı. p<0.05 değeri istatistiksel olarak anlamlı kabul edildi. Hepsinde lökosit ve trombosit sayıları azalmıştı. Hemoglobin değerinde farklılık görülmedi. Tanı ve takipteki CD45/SSC sonuçlarının karşılaştırılmasında granülosit ve blast yüzdelerinde azalma, lenfosit, monosit, normoblast yüzdelerinde artış saptandı. Hastaları MMR pozitif veya MMR negatif olarak 2 gruba ayırdığımızda tam kan sayımı ve CD45/SSC sonuçları farklı değildi. MMR pozitif ve negatif hastaların sağkalımı da benzer bulundu. Çalışmamızın sonuçlarına göre, CD45/SSC'nin rutin tanı ve/veya takip aracı olarak kullanılmasını öneremedik. Ancak çalışmamızın sınırlı hasta sayısı, MMR değerlendirme zamanı ile sadece bir zaman noktasındaki değerlendirme gibi kısıtlılıkları vardı. Seri CD45/SSC analizi ve farklı tirozin kinaz inhibitörleri ile yapılan daha büyük çalışmalarda sonuçlar değişebilir.

Anahtar Kelimeler

• kronik miyeloid lösemi
• CD45/SSC
• imatinib

Flow Cytometric Analysis of Chronic Phase Chronic Myeloid Leukemia Patients

Öz

Bone marrow aspirate for morphology and cytogenetics and qualitative reverse transcriptase polimerase chain reaction on peripheral blood cells is mandatory for the diagnosis of chronic myeloid leukemia (CML). Bone marrow biopsy and fluorescence in situ hybridisation may be necessary in selected patients. Performing other tests and diagnostic procedures depends on characteristics of the individual patient. Although flow cytometry is an essential tool in the diagnosis and monitoring of many hematological malignancies, it has a limited role in CML. In this study, we evaluated the CD45 side scatter results of our CML patients at diagnosis and during follow up. Totally 56 CML patients (22 female and 34 male) in chronic phase treated with imatinib were included. Patients were also evaluated after 8 (3-19) months follow up. Complete blood cound parameters and CD45/SSC results of the patients at diagnosis and follow up were evaluated retrospectively. The Wilcoxon T test was used to compare the means between the two groups. p<0.05 was considered statistically significant. All of them had decreased leukocyte and platelet counts. There was no difference in hemoglobin value. Comparison of CD45/SSC results at diagnosis and follow up revealed a decrease in granulocyte and blast percentages, and an increase in lymphocyte, monocyte, normoblast percentages. Complete blood counts and CD45/SSC results were not different when we divided patients into 2 groups according to being MMR positive or MMR negative. Survival of MMR positive and negative patients were also found similar. According to the results of our study, we were unable to suggest using CD45/SSC as a routine diagnostic and/or follow up tool. However, there were limitations of our study such as the limited number of patients, the variance between the time of MMR evaluation and the evaluation at only one time point. The results may change in larger studies with serial CD45/SSC analysis and with different tyrosine kinase inhibitors.

Anahtar Kelimeler

• chronic myeloid leukemia
• CD45/SSC
• imatinib

Kaynakça

1. 1. Buesche G, Hehlmann R, Hecker H, et al. Marrow fibrosis, indicator of therapy failure in chronic myeloid leukemia- prospective long-term results from a randomized-controlled trial. Leukemia. 2003;17:2444–53.
2. 2. Buesche G, Ganser A, Schlegelberger B, et al. Marrow fibrosis and its relevance during imatinib treatment of chronic myeloid leukemia. Leukemia. 2007;21:2420–7.
3. 3. Hidalgo-Lopez JE, Kanagal-Shamanna R, Quesada AE, et al. Bone marrow core biopsy in 508 consecutive patients with chronic myeloid leukemia: assessment of potential value. Cancer. 2018;124:3849–55.
4. 4. Hochhaus A, Baccarani M, Silver RT, et al. European LeukemiaNet 2020 recommendations for treating chronic myeloid leukemia. Leukemia 2020; 34:966–84.
5. 5. Sokal JE, Cox EB, Baccarani M, et al. Prognostic discrimination in “good-risk” chronic granulocytic leukemia. Blood. 1984;63:789–99.
6. 6. Hasford J, Pfirrmann M, Hehlmann R, et al. A new prognostic score for survival of patients with chronic myeloid leukemia treated with interferon alfa. Writing Committee for the Collaborative CML Prognostic Factors Project Group. J Natl Cancer Inst. 1998;90:850–8.
7. 7. Hasford J, Baccarani M, Hoffmann V, et al. Predicting complete cytogenetic response and subsequent progression-free survival in 2060 patients with CML on imatinib treatment: the EUTOS score. Blood. 2011;118:686–92.
8. 8. Pfirrmann M, Baccarani M, Saussele S, et al. Prognosis of long-term survival considering disease-specific death in patients with chronic myeloid leukemia. Leukemia. 2016;30:48–56.

9. 9. Dybko J, Haus O, Jaźwiec B, et al. CD117 (c-kit) expression on CD34+ cells participates in the cytogenetic response to imatinib in patients with chronic myeloid leukemia in the first chronic phase. Acta Haematol 2014;132:166–71.
10. 10. Shima H, Kiyokawa N, Miharu M, et al. Flow cytometric analysis as an additional predictive tool of treatment response in children with chronic-phase chronic myeloid leukemia treated with imatinib. Pediatr Blood Cancer 2017;64:264-78.
11. 11. Sun X, Li J, Chen J, et al. Flow cytometric assay of phosphotyrosine levels in Bcr-Abl-positive chronic myelogenous leukemias: a potential prognostic marker. Ann Hematol 2009; 88:29–36.
12. 12. Sviezhentseva IO, Perekhrestenk TP, Bilko DI, et al. Functional activity of CD34-positive cells in chronic myeloid leukemia patients with different response to imatinib therapy. Exp Oncol 2015; 37: 70–72.
13. 13. Oka S, Muroi K, Mori M, et al. Prediction of response to imatinib in patients with chronic myelogenous leukemia by flow cytometric analysis of bone marrow blastic cell phenotypes. Leukemia Lymphoma 2009; 50: 290–3.
14. 14.Stelzer GT, Shults KE, Loken MR. CD45 gating for routine flow cytometric analysis of human bone marrow specimens. Ann N Y Acad Sci 1993;677:265–80.
15. 15. Borowitz MJ, Guenther KL, Shults KE, et al. Immunophenotyping of acute leukemia by flow cytometricanalysis. Use of CD45 and right-angle light scatter to gate on leukemic blasts in three-color analysis. Am J Clin Pathol 1993;100:534–40.
16. 16. Janssen JJWM, Deenik W, Smolders KGM, et al. Residual normal stem cells can be detected in newly diagnosed chronic myeloid leukemia patients by a new flow cytometric approach and predict for optimal response to imatinib. Leukemia 2012; 26: 977-98.