RESUMO
INTRODUCTION: This analysis attempts to determine the diagnostic and prognostic value of bone marrow (BM) evaluation by multiparameter flow cytometry in patients with myelodysplastic syndrome (MDS). MATERIALS AND METHODS: The study group consisted of patients who underwent diagnostic process in the years 2008-2017 due to cytopenia and finally were diagnosed with MDS (n = 71). The comparative group included patients with cytopenia diagnosed in the same period, whose definitive diagnosis was other than MDS (n = 39). Flow cytometric evaluation of BM was performed following the recommendations of the European LeukemiaNet (ELN) in all patients. RESULTS: The median number of immunophenotypic abnormalities found on granulocytes in the MDS group was significantly higher compared to the comparative group [2 (range 0-5) vs 0 (range 0-2); P < .0001]. Similarly, the median Ogata score was significantly higher in the MDS group [2 (range 0-4) vs 1 (range 0-3); P < .0001]. Since the disturbances of the CD11b/HLA-DR and CD11b/CD13 on granulocytes were significantly more common in MDS patients, the Ogata score was extended by these abnormalities, what resulted in its higher diagnostic sensitivity (82%) while preserving high specificity (87%). The positive correlation was found between risk score determined by the Revised International Prognostic Scoring System and the number of the BM immunophenotypic abnormalities (P = .017). CONCLUSIONS: Our results indicate that the diagnostic usefulness of the Ogata score may be increased by including the abnormal expression of CD11b/HLA-DR and CD11b/CD13 on granulocytes. Moreover, our findings suggest the prognostic significance of the number of BM cytometric abnormalities in MDS.
Assuntos
Medula Óssea/patologia , Síndromes Mielodisplásicas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Medula Óssea/metabolismo , Antígeno CD11b/metabolismo , Antígenos CD13/metabolismo , Feminino , Citometria de Fluxo/métodos , Antígenos HLA-DR/metabolismo , Humanos , Imunofenotipagem/métodos , Masculino , Pessoa de Meia-Idade , Síndromes Mielodisplásicas/metabolismo , Prognóstico , Adulto JovemRESUMO
BACKGROUND: A flow cytometry score (FCS) based on four parameters has been proposed for analysis of myelodysplastic syndromes patients with <5% blasts. We evaluated whether bone marrow aspirate samples isolated from femoral heads could be used for verification of cutoff values of the individual parameters score (IPS), contributing to the FCS and compared the applicability of FCS parameters in two centers. STUDY DESIGN AND METHODS: Bone marrow cells were obtained from femoral heads of patients who underwent hip replacement surgery. The score of the 4 individual parameters of the cell samples were obtained independently in two centers, using their own facilities and methods. Flow cytometry data files were subsequently exchanged and reanalyzed in the other center. The resulting four data sets were compared to assess reproducibility and outcomes in both centers. RESULTS: Twenty-nine of 40 bone marrow samples contained sufficient cells for analysis. Proposed cutoff values for 3 of 4 individual parameters were appropriate. All 29 samples showed a positive individual parameters score (IPS:1) in 1 of the 4 obtained data sets. Most differences in IPS scores were a result of reanalyzing the data file in the other center, rather than data acquisition. FCS: ≥2 were observed in 11 samples. CONCLUSION: Bone marrow samples from femoral heads are appropriate for verification of the proposed reference cutoff values of the individual parameters. Proposed reference cutoff values needed to be adjusted for reliable interpretation. Standardization of both data acquisition and data analysis is necessary for obtaining uniform results.
Assuntos
Células da Medula Óssea/metabolismo , Cabeça do Fêmur/metabolismo , Citometria de Fluxo/métodos , Citometria de Fluxo/normas , Síndromes Mielodisplásicas/diagnóstico , Síndromes Mielodisplásicas/metabolismo , Idoso , Idoso de 80 Anos ou mais , Células da Medula Óssea/patologia , Feminino , Cabeça do Fêmur/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Síndromes Mielodisplásicas/patologiaRESUMO
BACKGROUND: Since the introduction of the orphan drugs legislation in Europe, it has been suggested that the general method of assessing drugs for reimbursement is not necessarily suitable for orphan drugs. The National Institute for Health and Clinical Excellence indicated that several criteria other than cost and efficacy could be considered in reimbursement decisions for orphan drugs. This study sought to explore the multi-criteria decision analysis (MCDA) framework proposed by (Orphanet J Rare Dis 7:74, 2012) to a range of orphan drugs, with a view to comparing the aggregate scores to the average annual cost per patient for each product, and thus establishing the merit of MCDA as a tool for assessing the value of orphan drugs in relation to their pricings. METHODS: An MCDA framework was developed using the nine criteria proposed by (Orphanet J Rare Dis 7:74, 2012) for the evaluation of orphan drugs, using the suggested numerical scoring system on a scale of 1 to 3 for each criterion. Correlations between the average annual cost of the drugs and aggregate MCDA scores were tested and plotted graphically. Different weightings for each of the attributes were also tested. A further analysis was conducted to test the impact of including the drug cost as an attribute in the aggregate index scores. RESULTS: In the drugs studied, the R 2, that statistically measures how close the data are to the fitted regression line was 0.79 suggesting a strong correlation between the drug scores and the average annual cost per patient. CONCLUSION: Despite several limitations of the proposed model, this quantitative study provided insight into using MCDA and its relationship to the average annual costs of the products.
Assuntos
Técnicas de Apoio para a Decisão , Produção de Droga sem Interesse Comercial , Descoberta de Drogas , Hemoglobinúria Paroxística/tratamento farmacológico , Hemoglobinúria Paroxística/metabolismo , Hipertensão Pulmonar/tratamento farmacológico , Hipertensão Pulmonar/metabolismo , Síndrome de Lennox-Gastaut/tratamento farmacológico , Síndrome de Lennox-Gastaut/metabolismo , Mucopolissacaridose II/tratamento farmacológico , Mucopolissacaridose II/metabolismo , Síndromes Mielodisplásicas/tratamento farmacológico , Síndromes Mielodisplásicas/metabolismoRESUMO
AIMS: In recent years, multiparameter flow cytometry has been increasingly recognised as an important tool in diagnosis of myelodysplastic syndrome and acute myeloid leukaemia (AML). Assessment of myeloid and monocytic 'immunophenotypic' dysplasia by flow cytometry in de novo AML has not been evaluated. METHODS: 97 cases of de novo AML cases were identified and reviewed by three hematopathologists. 'Immunophenotypic' dysplasia was assessed on blasts, monocytes and granulocytes by mean fluorescence intensity. RESULTS: Using the 2008 WHO classification criteria, there were 53 AML-not otherwise specified (NOS) (55%) and 28 AML with myelodysplasia-related changes (AML-MRC) (29%), while 16 cases were ambiguous as to AML-MRC status due to limited maturing cells for morphologic but adequate events number for immunophenotypic evaluation (AML-not evaluable, 16%). Compared with AML-NOS, granulocytic cells in AML-MRC had higher CD33 expression but lower CD45, CD11b and CD15. Monocytes in AML-MRC had lower expression of CD14, CD56 and CD45. Morphologic dysplasia was associated with significantly lower granulocytic forward scatter, side scatter and CD10 but higher CD33 expression. CONCLUSIONS: Our results suggest that the workup of AML cases should include flow cytometric assessment of granulocytes and monocytes. This analysis can aid a morphologic impression of multilineage dysplasia in distinguishing AML-MRC from AML-NOS, especially in cases with limited maturing myeloid cells.
Assuntos
Leucemia Mieloide Aguda/diagnóstico , Síndromes Mielodisplásicas/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Citometria de Fluxo , Humanos , Imunofenotipagem , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patologia , Leucócitos/metabolismo , Leucócitos/patologia , Masculino , Pessoa de Meia-Idade , Monócitos/metabolismo , Monócitos/patologia , Síndromes Mielodisplásicas/metabolismo , Síndromes Mielodisplásicas/patologia , Adulto JovemRESUMO
INTRODUCTION: The diagnosis of myelodysplastic syndrome (MDS) based on morphology is particularly difficult in low-grade MDS. Thus, the role of myeloid nuclear differentiation antigen (MNDA) and other flow cytometric (FCM) parameters in MDS was evaluated. METHODS: Bone marrow aspirates (BMA) collected from 52 patients with unexplained persistent cytopenias were divided into three groups: (i) proven MDS (n = 12) based on morphology and/or cytogenetics; (ii) suspected MDS (n = 6), noncontributory morphology, and cytogenetics; and (iii) non-MDS (n = 34). Sixteen control BMA were studied. Cases were analyzed for MNDA expression (on granulocytes, blasts, monocytes, and lymphocytes) and for seven quantitative parameters: CD34(+) myeloblasts % in nucleated cells, CD34(+) B-cell progenitor% in CD34(+) cells, lymphocyte/myeloblast CD45 MFI ratio, granulocyte/lymphocyte SSC peak channel ratio and the proportion of CD34(+) myeloblasts expressing CD15, CD11b, and CD56. A score of 1 was given to each parameter beyond the cutoff, and score ≥3 was considered FCM positive. RESULTS: MNDA expression on granulocytes and blasts was significantly lower in proven MDS and suspected MDS vs. non-MDS. Quantitative FCM parameters successfully distinguished MDS and suspected MDS from non-MDS. CONCLUSION: MNDA expression is an independent marker for the evaluation of dyspoiesis and may be added to the standard panel for quantitative assessment by FCM.
Assuntos
Antígenos de Diferenciação Mielomonocítica/metabolismo , Biomarcadores , Imunofenotipagem , Síndromes Mielodisplásicas/diagnóstico , Síndromes Mielodisplásicas/metabolismo , Fatores de Transcrição/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Medula Óssea/metabolismo , Medula Óssea/patologia , Células da Medula Óssea/metabolismo , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Curva ROCRESUMO
Although numerous recent publications have demonstrated interest in multiparameter flow cytometry in the investigation of myelodysplastic disorders, it is perceived by many laboratory hematologists as difficult and expensive, requiring a high level of expertise. We report a multicentric open real-life study aimed at evaluating the added value of the technically simple flow cytometry score described by the Ogata group for the diagnosis of myelodysplastic syndromes. A total of 652 patients were recruited prospectively in four different centers: 346 myelodysplastic syndromes, 53 myelodysplastic/myeloproliferative neoplasms, and 253 controls. The Ogata score was assessed using CD45 and CD34 staining, with the addition of CD10 and CD19. Moreover, labeling of CD5, CD7 and CD56 for the evaluation of myeloid progenitors and monocytes was tested on a subset of 294 patients. On the whole series, the specificity of Ogata score reached 89%. Respective sensitivities were 54% for low-risk myelodysplastic syndromes, 68% and 84% for type 1 and type 2 refractory anemia with excess of blasts, and 72% for myelodysplastic/myeloproliferative neoplasms. CD5 expression was poorly informative. When adding CD56 or CD7 labeling to the Ogata score, sensitivity rose to 66% for low-risk myelodysplastic syndromes, to 89% for myelodysplastic/myeloproliferative neoplasms and to 97% for refractory anemia with excess of blasts. This large multicenter study confirms the feasibility of Ogata scoring in routine flow cytometry diagnosis but highlights its poor sensitivity in low-risk myelodysplastic syndromes. The addition of CD7 and CD56 in flow cytometry panels improves the sensitivity but more sophisticated panels would be more informative.
Assuntos
Antígenos CD7/metabolismo , Antígenos CD5/metabolismo , Antígeno CD56/metabolismo , Imunofenotipagem , Síndromes Mielodisplásicas/diagnóstico , Síndromes Mielodisplásicas/metabolismo , Doenças Mieloproliferativas-Mielodisplásicas/diagnóstico , Doenças Mieloproliferativas-Mielodisplásicas/metabolismo , Idoso , Idoso de 80 Anos ou mais , Antígenos CD7/genética , Antígenos CD5/genética , Antígeno CD56/genética , Diagnóstico Diferencial , Citometria de Fluxo , Expressão Gênica , Humanos , Imunofenotipagem/métodos , Pessoa de Meia-Idade , Síndromes Mielodisplásicas/genética , Doenças Mieloproliferativas-Mielodisplásicas/genética , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Myeloid nuclear differentiation antigen (MNDA) is expressed in myelomonocytic cells with highest levels in mature granulocytes and monocytes. It is suggested to be expressed more weakly in patients with myelodysplastic syndromes (MDS). The analysis of MNDA therefore may improve diagnostic capabilities of multiparameter flow cytometry (MFC) in MDS. METHODS: We used MFC for detection of MNDA expression in 269 patients with suspected or known MDS, acute myeloid leukemia (AML) or chronic myelomonocytic leukemia (CMML), cytopenia of unknown cause or without malignancy (negative controls). Results were compared with the diagnoses revealed by cytomorphology (CM) and cytogenetics (CG). RESULTS: Percentages of granulocytes and monocytes with diminished MNDA expression (dimG and dimM) were higher in patients with MDS (mean ± SD, 20% ± 20%, P < 0.001 and 31% ± 24%, P < 0.001) and AML (27% ± 27%, P = 0.007 and 45% ± 31%, P = 0.001) diagnosed by CM, vs. patients without MDS (8% ± 10% and 16% ± 11%), respectively. Significant differences were also found for mean fluorescence intensity (MFI) of MNDA in monocytes which was lower in MDS (mean ± SD, 71 ± 36, P = 0.004) and AML (55 ± 39, P < 0.001) vs. no MDS samples (85 ± 28), respectively. Within patients with MDS, cases with cytogenetic aberrations showed a trend to higher %dimG (24% ± 18%, P = 0.083) compared with those without (16% ± 21%). Cut-off values for %dimG (12%) and %dimM (22%) as well as for MFI in monocytes (72) were defined capable of discriminating between MDS and non-MDS. CONCLUSION: MNDA expression in bone marrow cells can be assessed reliably by MFC and may facilitate evaluation of dyspoiesis when added to a standard MDS MFC panel.
Assuntos
Antígenos de Diferenciação Mielomonocítica/metabolismo , Biomarcadores Tumorais/metabolismo , Síndromes Mielodisplásicas/metabolismo , Fatores de Transcrição/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Feminino , Citometria de Fluxo , Granulócitos/metabolismo , Humanos , Linfócitos/metabolismo , Masculino , Pessoa de Meia-Idade , Monócitos/metabolismo , Síndromes Mielodisplásicas/diagnóstico , Síndromes Mielodisplásicas/patologia , Células Progenitoras Mieloides/metabolismo , Valores de Referência , Adulto JovemRESUMO
By using flow cytometry, we analyzed myeloid nuclear differentiation antigen (MNDA) expression in myeloid precursors in bone marrow from patients with myelodysplastic syndrome (MDS) and control samples from patients undergoing orthopedic procedures. The median percentage of MNDA-dim myeloid precursors in MDS cases was 67.4% (range, 0.7%-97.5%; interquartile range, 44.9%-82.7%) of myeloid cells, with bimodal MNDA expression in most MDS samples. Control samples demonstrated a median MNDA-dim percentage in myeloid precursors of 1.2% (range, 0.2%-13.7%; interquartile range, 0.6%-2.7%), with no bimodal pattern in most samples. The area under the receiver operating characteristic curve for MNDA-dim percentage in myeloid precursors was 0.96 (P = 9 × 10(-7)). Correlation of MNDA-dim levels with World Health Organization 2008 morphologic diagnoses was not significant (P = .21), but correlation with patient International Prognostic Scoring System scores suggested a trend (P = .07). Flow cytometric assessment of MNDA in myeloid precursors in bone marrow may be useful for the diagnosis of MDS.