Combined flow cytometric assessment of CD45, HLA‐DR, CD34, and CD117 expression is a useful approach for reliable quantification of blast cells in myelodysplastic syndromes

Sandes, Alex Freire; Kerbauy, Daniela Márcia Bahia; Matarraz, Sergio; Chauffaille, Maria de Lourdes Lopes Ferrari; López, Antonio; Órfão, Alberto; Yamamoto, Masahiro

doi:10.1002/cyto.b.21087

Cited by 34 publications

(35 citation statements)

References 31 publications

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“…The frequency CD34+CD45 dim SSC low and CD45 dim SSC low cell populations is usually lower than the number of blasts by morphology [13,17], while addition of CD117+/CD45 dim /SSC low population to CD34+ cells will overestimate blast frequency due to inclusion of erythroid precursors [11,46]. A recent study compared four different FCM strategies with morphological blast counting and concluded that quantification of "CD34+ and/or CD117+HLA-DR+" cells (derived from all nucleated BM cells) is the most efficient strategy to evaluate blasts in both normal/reactive BM and in the BM from MDS patients [70]. By this strategy, the mean overall difference between blast frequency by FCM and morphology was 0.2 %, while CD34+CD45 dim strategy was not sensitive enough [70].…”

Section: Can Flow Cytometry Add To Risk Assessment Of Mds Patients?mentioning

confidence: 96%

“…A recent study compared four different FCM strategies with morphological blast counting and concluded that quantification of "CD34+ and/or CD117+HLA-DR+" cells (derived from all nucleated BM cells) is the most efficient strategy to evaluate blasts in both normal/reactive BM and in the BM from MDS patients [70]. By this strategy, the mean overall difference between blast frequency by FCM and morphology was 0.2 %, while CD34+CD45 dim strategy was not sensitive enough [70]. Further studies are needed to validate clinical significance of this strategy in a larger material.…”

Section: Can Flow Cytometry Add To Risk Assessment Of Mds Patients?mentioning

confidence: 99%

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Is There a Role for Flow Cytometry in the Evaluation of Patients With Myelodysplastic Syndromes?

Porwit

2015

Curr Hematol Malig Rep

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This review focuses on the most recent literature concerning flow cytometry (FCM) application for diagnosis of myelodysplastic syndrome (MDS). Aberrant FCM results have been defined as abnormalities in at least three tested features comprising at least two bone marrow (BM) cell compartments. FCM results should be interpreted together with the BM smear cytology, the morphological assessment of BM biopsy, and cytogenetic results. Including FCM in the pre-treatment assessment may provide not only diagnostic but also prognostic information. Further studies are needed to evaluate the role of FCM in individual risk assessment for MDS patients and in therapy choice and/or follow-up.

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Section: Can Flow Cytometry Add To Risk Assessment Of Mds Patients?mentioning

confidence: 96%

Section: Can Flow Cytometry Add To Risk Assessment Of Mds Patients?mentioning

confidence: 99%

Is There a Role for Flow Cytometry in the Evaluation of Patients With Myelodysplastic Syndromes?

Porwit

2015

Curr Hematol Malig Rep

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“…Monoclonal antibodies that were used in this study included fluorescein (17,19). B cell progenitors were discriminated from myeloid progenitors by lower CD45, lower SSC properties and back gating with CD19 and were excluded from myeloid progenitor analysis (17,18).…”

Section: Flow Cytometric Analysis Of Bone Marrow Samplesmentioning

confidence: 99%

Absence of aberrant myeloid progenitors by flow cytometry is associated with favorable response to azacitidine in higher risk myelodysplastic syndromes

et al. 2014

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Background: In intermediate-2 and high risk patients with myelodysplastic syndromes (MDS), treatment with azacitidine is associated with hematological responses and prolonged overall survival in patients who respond to therapy. However, only half of the patients that are treated will benefit from this treatment. It is a major challenge to predict which patients are likely to respond to treatment. The aim of this study was to investigate the predictive value of immunophenotyping for response to treatment with azacitidine of Int-2 and high risk MDS patients. Methods: Bone marrow aspirates were analyzed by flow cytometry in 42 patients with Int-2 and high risk MDS, chronic myelomonocytic leukemia or low blast count acute myeloid leukemia before treatment and after every third cycle of azacitidine. A flow score was calculated using the flow cytometric scoring system (FCSS). Results: The presence of myeloid progenitors with an aberrant immunophenotype was significantly associated with lack of response (p=0.02). A low pretreatment FCSS was associated with significantly better overall survival compared with a high pretreatment FCSS (p=0.03). A significant decrease in FCSS was observed in patients with complete response after three cycles azacitidine compared to patients with progressive disease (p=0.006). Conclusions: Absence of aberrant myeloid progenitor cells at baseline and/or a decrease in the FCSS during treatment identified Int-2 and high risk MDS patients who are likely to respond to treatment with azacitidine. © 2014 Clinical Cytometry Society.

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“…For detailed calculations of frequency of GPI-AP deficiency please review citation [13]. *Leukemic and non-leukemic cells were sorted using the following markers: HLA-DR, CD13, CD117 and CD45 as described earlier with some modifications [54]. …”

Section: Resultsmentioning

confidence: 99%

“…Leukemic blasts were sorted from CD34+ cells under BSL-2 conditions with a 16-color BD FACSAria SORP high speed cell sorter (Becton Dickinson) using the following markers: HLA-DR, CD13, CD117 and CD45 as previously described with few modifications [54]. …”

Section: Methodsmentioning

confidence: 99%

PIGN gene expression aberration is associated with genomic instability and leukemic progression in acute myeloid leukemia with myelodysplastic features

et al. 2017

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Previous studies have linked increased frequency of glycosylphosphatidylinositol-anchor protein (GPI-AP) deficiency with genomic instability and the risk of carcinogenesis. However, the underlying mechanism is still not clear. A randomForest analysis of the gene expression array data from 55 MDS patients (GSE4619) demonstrated a significant (p = 0.0007) correlation (Pearson r =-0.4068) between GPI-anchor biosynthesis gene expression and genomic instability, in which PIGN, a gene participating in GPI-AP biosynthesis, was ranked as the third most important in predicting risk of MDS progression. Furthermore, we observed that PIGN gene expression aberrations (increased transcriptional activity but diminished to no protein production) were associated with increased frequency of GPI-AP deficiency in leukemic cells during leukemic transformation/progression. PIGN gene expression aberrations were attributed to partial intron retentions between exons 14 and 15 resulting in frameshifts and premature termination which were confirmed by examining the RNA-seq data from a group of AML patients (phs001027.v1.p1). PIGN gene expression aberration correlated with the elevation of genomic instability marker expression that was independent of the TP53 regulatory pathway. Suppression/elimination of PIGN protein expression caused a similar pattern of genomic instability that was rescued by PIGN restoration. Finally, we found that PIGN bound to the spindle assembly checkpoint protein, MAD1, and regulated its expression during the cell cycle. In conclusion, PIGN gene is crucial in regulating mitotic integrity to maintain chromosomal stability and prevents leukemic transformation/progression.

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Combined flow cytometric assessment of CD45, HLA‐DR, CD34, and CD117 expression is a useful approach for reliable quantification of blast cells in myelodysplastic syndromes

Cited by 34 publications

References 31 publications

Is There a Role for Flow Cytometry in the Evaluation of Patients With Myelodysplastic Syndromes?

Is There a Role for Flow Cytometry in the Evaluation of Patients With Myelodysplastic Syndromes?

Absence of aberrant myeloid progenitors by flow cytometry is associated with favorable response to azacitidine in higher risk myelodysplastic syndromes

PIGN gene expression aberration is associated with genomic instability and leukemic progression in acute myeloid leukemia with myelodysplastic features

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