Chu S.-C.,Buddhist Tzu Chi General Hospital |
Chu S.-C.,Institute of Medical science |
Wang T.-F.,Buddhist Tzu Chi General Hospital |
Li C.-C.,National Taiwan University Hospital |
And 3 more authors.
Leukemia Research | Year: 2011
The aim of this study is to validate the clinical utility of the flow cytometric scoring system (FCSS), quantifying phenotypic aberrancies in the myelomonocytic lineages, in the diagnosis and prognosis for conventionally treated myelodysplastic syndromes (MDS) patients. The bone marrow samples from 56 consecutive newly diagnosed MDS patients were characterized by the FCSS and compared with findings in 27 non-MDS cytopenic patients. The FCSS scores were significantly higher in patients with MDS than those in the non-MDS control. A flow score of 2 or more allowed for a specificity of 100% with 75% sensitivity in distinguishing these two groups. The FCSS scores correlated directly with validated prognostic systems including WHO classification, International Prognostic Scoring System (IPSS), WHO-adjusted prognostic scoring system (WPSS) and transfusion dependency. The median survival of conventionally treated MDS patients was directly related to FCSS group; severe: 6. months; moderate: 19. months and normal/mild: not reached. The multivariate analyses suggested the FCSS risk categories were an independent prognostic factor after adjustment for sex, age (above or below 70. years), IPSS or WPSS risk categories. These results confirm that quantifying aberrancies in the myelomonocytic lineage by FCSS is useful in MDS diagnosis and extends the prognostic utility for conventionally treated/untreated patients, especially among patients classified within the refractory cytopenia with multilineage dysplasia (RCMD) subgroup. © 2011 Elsevier Ltd.
Cooper T.M.,Emory University |
Alonzo T.A.,University of Southern California |
Gerbing R.B.,Childrens Oncology Group |
Perentesis J.P.,Cincinnati Childrens Medical Center |
And 7 more authors.
Cancer | Year: 2014
BACKGROUND The discovery of new, effective non-anthracycline-based reinduction regimens for children with recurrent acute myeloid leukemia (AML) is critical. In this phase 1/2 study, the tolerability and overall response rate of clofarabine in combination with cytarabine was investigated in children with recurrent/refractory AML. METHODS AAML0523 enrolled 49 children with AML in first recurrence or who were refractory to induction therapy. The study consisted of a dose-finding phase (9 patients) and an efficacy phase (40 patients). Two children received clofarabine at a dose of 40 mg/m 2/day and 47 children at a dose of 52 mg/m2/day. RESULTS Toxicities typical for intensive chemotherapy regimens were observed at all doses of clofarabine. The recommended pediatric phase 2 dose of clofarabine in combination with cytarabine was 52 mg/m2/day for 5 days. Of 48 evaluable patients, the overall response rate (complete remission plus complete remission with partial platelet recovery) was 48%. Four patients met conventional criteria for complete remission with incomplete count recovery. Twenty-one of 23 responders subsequently underwent hematopoietic stem cell transplantation. The overall survival rate at 3 years was 46% for responders compared with 16% for nonresponders (P < .001). Patients found to have no minimal residual disease at the end of the first cycle by flow cytometric analysis had superior overall survival after 1 year (100% vs 38%; P = .01). CONCLUSIONS The combination of clofarabine and cytarabine yielded an acceptable response rate without excess toxicity in children with recurrent AML. The nearly 50% survival rate reported in responders is highly encouraging in these high-risk patients and suggests that this combination is an effective bridge to hematopoietic stem cell transplantation. © 2014 American Cancer Society.
Porwit A.,University of Toronto |
Van De Loosdrecht A.A.,VU University Amsterdam |
Bettelheim P.,Elisabethinen Hospital |
Eidenschink Brodersen L.,HematoLogics Inc. |
And 17 more authors.
Leukemia | Year: 2014
Definite progress has been made in the exploration of myelodysplastic syndromes (MDS) by flow cytometry (FCM) since the publication of the World Health Organization 2008 classification of myeloid neoplasms. An international working party initiated within the European LeukemiaNet and extended to include members from Australia, Canada, Japan, Taiwan and the United States has, through several workshops, developed and subsequently published consensus recommendations. The latter deal with preanalytical precautions, and propose small and large panels, which allow evaluating immunophenotypic anomalies and calculating myelodysplasia scores. The current paper provides guidelines that strongly recommend the integration of FCM data with other diagnostic tools in the diagnostic work-up of MDS. © 2014 Macmillan Publishers Limited. All rights reserved.
Ostronoff F.,Fred Hutchinson Cancer Research Center |
Ostronoff F.,University of Washington |
Othus M.,Fred Hutchinson Cancer Research Center |
Othus M.,Statistical Center |
And 16 more authors.
Blood | Year: 2014
NUP98/NSD1 has recently been reported in association with poor outcome in acute myeloid leukemia (AML). Previous studies also observed a high overlap between NUP98/NSD1 and FLT3/ITD, raising the question as to whether the reported poor outcome is due to NUP98/NSD1 or caused by the co-occurrence of these 2 genetic lesions. We aimed to determine the prognostic significance of NUP98/NSD1 in the context of FLT3/ITD AML. A total of 1421 patients enrolled in 5 consecutive Children's Oncology Group/Children's Cancer Group and SWOG trials were evaluated. NUP98/NSD1 was found in 15% of FLT3/ITD and 7% of cytogenetically normal (CN)-AML. Those with dual FLT3/ITD and NUP98/NSD1 (82% of NUP98/NSD1 patients) had a complete remission rate of 27% vs 69% in FLT3/ITD without NUP98/NSD1 (P < .001). The corresponding 3-year overall survival was 31% vs 48% (P = .011), respectively. In CN-AML, patients with concomitant NUP98/NSD1 and FLT3/ITD had a worse outcome than those harboring NUP98/NSD1 only. In multivariate analysis, the dual NUP98/NSD1 and FLT3/ITD remained an independent predictor of poor outcome, and NUP98/NSD1 without FLT3/ITD lost its prognostic significance. Our study demonstrates that it is the interaction between NUP98/NSD1 and FLT3/ITD that determines the poor outcome of patients with NUP98/NSD1 disease. © 2014 by The American Society of Hematology.
Loken M.R.,Hematologics Inc. |
Alonzo T.A.,University of Southern California |
Alonzo T.A.,Childrens Oncology Group |
Pardo L.,Fundaleu Hospital |
And 14 more authors.
Blood | Year: 2012
Early response to induction chemotherapy is a predictor of outcome in acute myeloid leukemia (AML). We determined the prevalence and significance of postinduction residual disease (RD) by multidimensional flow cytometry (MDF) in children treated on Children's Oncology Group AML protocol AAML03P1. Postinduction marrow specimens at the end of induction (EOI) 1 or 2 or at the end of therapy from 249 patients were prospectively evaluated by MDF for RD, and presence of RD was correlated with disease characteristics and clinical outcome. Of the 188 patients in morphologic complete remission at EOI1, 46 (24%) had MDFdetectable disease. Those with and without RD at the EOI1 had a 3-year relapse risk of 60% and 29%, respectively (P < .001); the corresponding relapsefree survival was 30% and 65% (P < .001). Presence of RD at the EOI2 and end of therapy was similarly predictive of poor outcome. RD was detected in 28% of standard-risk patients in complete remission and was highly associated with poor relapse-free survival (P = .008). In a multivariate analysis, including cytogenetic and molecular risk factors, RD was an independent predictor of relapse (P < .001). MDF identifies patients at risk of relapse and poor outcome and can be incorporated into clinical trials for riskbased therapy allocation. This study was registered at www.clinicaltrials.gov as NCT00070174. © 2012 by The American Society of Hematology.
PubMed | University of Minnesota, The Hospital for Sick Children, Fred Hutchinson Cancer Research Center, Maine Medical Center and 6 more.
Type: Journal Article | Journal: Clinical cancer research : an official journal of the American Association for Cancer Research | Year: 2016
Gemtuzumab ozogamicin (GO), a calicheamicin-conjugated mAb against CD33, has been used in the treatment of acute myeloid leukemia (AML). We evaluated the impact of the addition of GO to standard chemotherapy and hematopoietic stem cell transplant (HCT) in patients withFLT3/ITD.We analyzed children withFLT3/ITD-positive AML (n= 183) treated on two consecutive Childrens Oncology Group AML trials (NCT00070174andNCT00372593). Outcomes were assessed forFLT3/ITD patients receiving standard chemotherapy with or without GO (GO vs. No-GO, respectively), and the impact of consolidation HCT for high-riskFLT3/ITD patients [highFLT3/ITD allelic ratio (ITD-AR)].For allFLT3/ITD patients, complete remission (CR) rates for the GO versus No-GO cohorts were identical (64% vs. 64%;P= 0.98). Relapse rate (RR) after initial CR was 37% for GO recipients versus 59% for No-GO recipients (P= 0.02), disease-free survival (DFS) was similar (47% vs. 41%;P= 0.45), with higher treatment-related mortality (TRM) in GO recipients (16% vs. 0%;P= 0.008). Among high-riskFLT3/ITD patients with high ITD-AR, those who received HCT in first CR with prior exposure to GO had a significant reduction in RR (15% vs. 53%;P= 0.007), with a corresponding DFS of 65% versus 40% (P= 0.08), and higher TRM (19% vs. 7%;P= 0.08).CD33 targeting with HCT consolidation may be an important therapeutic strategy in high-riskFLT3/ITD AML and its efficacy and associated toxicity warrant further investigation.
PubMed | Fred Hutchinson Cancer Research Center and HematoLogics Inc.
Type: Journal Article | Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology | Year: 2016
Identification and quantification of maturing hematopoietic cell populations in flow cytometry data sets is a complex and sometimes irreproducible step in data analysis. Supervised machine learning algorithms present promise to automatically classify cells into populations, reducing subjective bias in data analysis. We describe the use of support vector machines (SVMs), a supervised algorithm, to reproducibly identify two distinctly different populations of normal hematopoietic cells, mature lymphocytes and uncommitted progenitor cells, in the challenging setting of pediatric bone marrow specimens obtained 1 month after chemotherapy. Four-color flow cytometry data were collected on a FACS Calibur for 77 randomly selected postchemotherapy pediatric patients enrolled on the Childrens Oncology Group clinical trial AAML1031. These patients demonstrated no evidence of detectable residual disease and were divided into training (n=27) and testing (n=50) cohorts. SVMs were trained to identify mature lymphocytes and uncommitted progenitor cells in the training cohort before independent evaluation of prediction efficiency in the testing cohort. Both SVMs demonstrated high predictive performance (lymphocyte SVM: sensitivity >0.99, specificity >0.99; uncommitted progenitor cell SVM: sensitivity=0.94, specificity >0.99) and closely mirrored manual cell classifications by two expert-analysts. SVMs present an efficient, automated methodology for identifying normal cell populations even in stressed bone marrows, replicating the performance of an expert while reducing the intrinsic bias of gating procedures between multiple analysts. 2016 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of ISAC.
Wangen J.R.,Hematologics Inc. |
Eidenschink Brodersen L.,Hematologics Inc. |
Stolk T.T.,VU University Amsterdam |
Wells D.A.,Hematologics Inc. |
Loken M.R.,Hematologics Inc.
International Journal of Laboratory Hematology | Year: 2014
Introduction: The extension of quantitative flow cytometric studies to the erythroid lineage in patients with suspected myelodysplastic syndrome has prompted a reassessment of cell surface antigen expression during normal erythropoiesis. Erythropoiesis in normal and pathologic bone marrows was studied to determine the expected antigenic relationships of maturing erythroid cells. Methods: A total of 200 bone marrow specimens were evaluated by multidimensional flow cytometry (MDF). Samples were prepared using either NH4Cl lysis or Ficoll density gradient separation. Results: Normal erythroid development is described as a two-step process observable with the intensity relationships between CD235a, CD71, CD45, CD105, CD34, CD117, and CD36. The variability of these intensities (CV) was determined. A comparison of processing techniques determined lysis is the optimal analytic technique for the analysis of early-stage erythroid cells. Nucleic acid staining with DRAQ5 revealed that Ficoll allows for the analysis of reticulocytes and mature erythrocytes otherwise eliminated by lysis. Conclusion: These data demonstrate while lysis alters the light scatter characteristics of erythroid precursors, it did not alter quantitative antigen expression or nucleic acid content. The expected variability in antigen intensities is defined. These studies provide a basis for a comparison of erythroid development between normal individuals and those with erythroid dysplasia associated with myelodysplastic syndromes. © 2013 John Wiley & Sons Ltd.
Foster J.H.,Baylor College of Medicine |
Hawkins D.S.,Seattle Childrens |
Loken M.R.,HematoLogics Inc. |
Wells D.A.,HematoLogics Inc. |
Thomson B.,Seattle Childrens
Pediatric Blood and Cancer | Year: 2011
Previous studies to evaluate minimal disease in acute lymphoblastic leukemia (ALL) after treatment have relied on the diagnostic specimen to develop patient-specific analytical probes. The diagnostic specimen is often not available in a tertiary setting; therefore, we evaluated the use of flow cytometry (FCM) using a "difference from normal" approach to detect residual disease prior to myeloablative allogeneic hematopoietic cell transplantation (HCT). Among 116 pediatric patients with ALL who were in morphological remission at time of transplant, we found that those patients who had detectable residual disease by FCM prior to HCT experienced significantly inferior outcome. © 2011 Wiley-Liss, Inc.
Hematologics Inc. | Date: 2013-12-05
A system, method, and article for diagnosing a test set of biological cells. For example, in one embodiment a normal set of cells is characterized using flow cytometry. A centroid and radius are defined for a set of clusters in an n-dimensional space corresponding to a normal maturation for a cell lineage in the normal set of cells. A test set of cells is characterized using flow cytometry and the characterization is compared to the set of clusters.