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Micro RNAs (miRNAs) expressions and flow cytometry immunophenotyping (FCI) in myelodisplastic syndrome (MDS) diagnosed patients - are they optional diagnostic methods?

Specific micro RNAs (miRNAs) that are unique to myelodisplastic syndrome (MDS) have emerged as critical regulators of hematopoiesis and recent findings have implicated their aberrant expression in MDS. In 2017 MDS Working Group of the Croatian Cooperative Group for Hematologic Diseases (CROHEM), Referral center of the Ministry of Health of the Republic of Croatia as well as the Croatian Society for Haematology of the Croatian Medical Association published Croatian guidelines for diagnosis and treatment of MDS based on morphological characteristics of hematopoietic cells supplemented with the cytogenetic analysis and bone marrow (BM) flow cytometry immunophenotyping (FCI) method. To investigate weather the aberrant expression of some specific miRNAs and FCI method could be only the optional diagnostic methods in diagnosis of MDS. After the MDS diagnosis according to Croatian guidelines was set, 62 BM samples (diagnosed as MDS by histopathology) were processed to multicolor/multiparameter analysis of dysplasia with the FCI method ISO 15189 accredited according to the European LeukemiaNet (ELN) Working Group protocols on FC 500 (Beckman Coulter). Circulating miRNAs (miR- 125a and miR-99b) aberrant expression was performed (plasma samples from 4 healthy volunteers and 21 MDS, miScript Custom PCR Array, Qiagen) on TaqMan qPCR platform in a molecular laboratory accredited according ISO 15189. In BM samples of 62 MDS to 9 phenotypic anomalies were found – 50 were FCI positive MDS, 5 FCI suspected MDS and 7 negative FCI MDS. No statistically significant differences of miRNAs expressions between healthy volunteers and MDS were observed (miR-125a = 0, 4139 ; miR-99b = 0, 3580), but both miRNAs had aberant expression, were up- regulated in MDS (miR-125a =1.39 ; miR- 99b = 1.74), but still not over 2-fold change of healthy volunteers. As it is presented, the expression of both miRNAs is aberrantly changed and their even so small up- regulated expressions could be useful for further diagnostic purposes in MDS. In the future, extended Croatian miRNAs profiling is planned with let 7a, miR-16, miR-21, miR-144, miR- 451, miR-651 and miR-655 wich, according to recent findings, could split the MDS patients into two groups with regards to the risk level. Although FCI method itself is not a basis for a MDS diagnosis, today it is an optional diagnostic method in the diagnosis and prognosis of MDS through multiparameter analysis data that are already incorporated into the 2016 revision to the World Health Organization (WHO) classification of myeloid neoplasms and acute leukemia.

miRNAs expressions ; FCI ; MDS

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