Residual disease detected by multidimensional flow cytometry signifies high relapse risk in patients with de novo acute myeloid leukemia: a report from Children's Oncology Group

Loken, Michael R.; Alonzo, Todd A.; Pardo, Laura; Gerbing, Robert B.; Raimondi, Susana C.; Hirsch, Betsy A.; Ho, Phoenix A.; Franklin, Janet; Cooper, Todd M.; Gamis, Alan S.; Meshinchi, Soheil

doi:10.1182/blood-2012-02-408336

Cited by 266 publications

(271 citation statements)

References 20 publications

Supporting

Mentioning

248

Contrasting

Unclassified

Order By: Relevance

“…39 Minimal residual disease detected using up to 10-color multidimensional flow cytometry has been shown to predict relapse in MAC allo-HCT settings in adults or in children. 40,41 Our study did not find that patients with lower blast counts had favorable outcome compared with those with higher blast counts, while still being in morphologic CR. This suggests that the quantity of non-leukemic myeloblasts has no influence on outcome.…”

Section: Discussionmentioning

confidence: 40%

Achieving stringent CR is essential before reduced-intensity conditioning allogeneic hematopoietic cell transplantation in AML

Üstün

Wiseman

DeFor

et al. 2013

Bone Marrow Transplant

View full text Add to dashboard Cite

Reduced-intensity conditioning (RIC) allogeneic hematopoietic cell transplantation (allo-HCT) can cure patients with AML in CR. However, relapse after RIC allo-HCT may indicate heterogeneity in the stringency of CR. Strict definition of CR requires no evidence of leukemia by both morphologic and flow cytometric criteria. We re-evaluated 85 AML patients receiving RIC allo-HCT in CR to test if a strict definition of CR had direct implications for the outcome. These patients had leukemia immunophenotype documented at diagnosis and analyzed at allo-HCT. Eight (9.4%) had persistent leukemia by flow cytometric criteria at allo-HCT. The patients with immunophenotypic persistent leukemia had a significantly increased relapse (hazard ratio (HR): 3.7; 95% confidence interval (CI): 1.3-10.3, P ¼ 0.01) and decreased survival (HR: 2.9; 95% CI: 1.3-6.4, Po0.01) versus 77 patients in CR by both morphology and flow cytometry. However, the pre-allo-HCT bone marrow (BM) blast count (that is, 0-4%) was not significantly associated with risks of relapse or survival. These data indicate the presence of leukemic cells, but not the BM blast count affects survival. A strict morphologic and clinical lab flow cytometric definition of CR predicts outcomes after RIC allo-HCT, and therefore is critical to achieve at transplantation. Keywords: allogeneic hematopoietic cell transplantation; reduced-intensity conditioning regimen; AML; relapse; minimal residual disease; CR INTRODUCTIONAllogeneic hematopoietic SCT (allo-HCT) remains the most effective treatment for most patients with AML. 1 The two main mechanisms by which allo-HCT can cure AML are through the immunologically based GVL effect and leukemic cell cytoreduction by HCT conditioning. 2-5 Reduced-intensity conditioning (RIC) was introduced to allo-HCT more than a decade ago for patients who are older, had significant co-morbid conditions or poorer performance status. [6][7][8] The anti-neoplastic potency of these RIC HCT regimens relies primarily on the GVL effect rather than ablating all residual leukemic disease. 9 It is clear that patients with active leukemia had more relapse and worse OS after allo-HCT regardless of donor type or patient age. [10][11][12][13][14][15] Following allo-HCT, 40-60% of AML patients in CR1 enjoy long-term OS, whereas o20% of refractory or relapsed AML patients survive. 1,11 Yet even for allo-HCT during CR, relapse remains the most frequent complication of allo-HCT. 16 As relapse has been reported more frequently after RIC allo-HCT compared with myeloablative conditioning (MAC) allo-HCT in patients with AML, this suggests heterogeneity in the interpretation of a CR, which may be more important after RIC allo-HCT. [17][18][19][20][21][22][23] The CR definition updated in 2003 24 clearly requires no evidence of leukemia by flow cytometry in addition to the morphologic remission as reported: 'The presence of a unique phenotype (by flow cytometry) identical to what was found in the pretreatment specimen (for example, CD34, CD7 coexpression) should be viewed ...

show abstract

Section: Discussionmentioning

confidence: 40%

Achieving stringent CR is essential before reduced-intensity conditioning allogeneic hematopoietic cell transplantation in AML

Üstün

Wiseman

DeFor

et al. 2013

Bone Marrow Transplant

View full text Add to dashboard Cite

show abstract

“…The detection of abnormal cell populations based on “Difference from Normal” requires a precise knowledge of gene product expression during hematopoiesis 3. Defining normal gene product expression patterns is predicated on both consistent gene product expression from individual to individual and a stable analytic platform in which to detect cellular characteristics on stressed cells.…”

Section: Discussionmentioning

confidence: 99%

“…This approach patterns the normal regenerating bone marrow cells in multidimensional space in order to identify populations or clusters of events that are at least 0.5 log units disparate from the nearest normal counterpart 3, 4. Low levels of leukemia cells can thus be identified based on surface gene product expression patterns that are different from those observed in normal hematopoiesis.…”

Section: Introductionmentioning

confidence: 99%

Consistent quantitative gene product expression: #2. Antigen intensities on bone marrow cells are invariant between individuals

Loken¹,

Voigt²,

Brodersen³

et al. 2016

Cytometry Pt A

Self Cite

View full text Add to dashboard Cite

Five reference populations in bone marrow specimens were identified by flow cytometry using specific combinations of reagents in order define the variation of gene product expression intensities both within and between individuals. Mature lymphocytes, uncommitted progenitor cells, promyelocytes, mature monocytes and mature neutrophils can be reproducibly identified as distinct clusters of events in heterogeneous, maturing bone marrow specimens. Support Vector Machines were used to identify the reference populations in order to reduce subjective bias in manually defining boundaries of these populations since they were not discretely separated from the remainder of the cells. Reference populations were identified in 50 randomly selected bone marrow aspirates obtained over a period spanning 3 years and 6 months from pediatric patients following chemotherapy for acute myeloid leukemia (AML). The quantitative expression of gene products (cell surface antigens) and light scattering characteristics on these stressed specimens were demonstrated to be tightly regulated both within individuals and between individuals. Within an individual most gene products (CD45, CD34, CD14, CD16, CD64, CD33) demonstrated limited variability with a standard deviation of <0.20 log units while CD13 and CD36 exhibited broader variation >0.25 log units. Surprisingly, with the exception of CD33, the variation of the mean intensities of each antigen between individuals was even less than the variation within an individual. These data confirm that the amounts of gene products expressed on normal developing cells are highly regulated but differ in intensities between different lineages and during the maturational pathway of those lineages. The amounts of gene products expressed at specific stages of development of each lineage are a biologic constant with minimal variation within or between individuals. © 2016 The Authors. Cytometry Part A Published by Wiley Periodicals, Inc. on behalf of ISAC.

show abstract

“…For three separate Children's Oncology Group AML trials over a period of 13 years, standardized four‐color panels were used to pattern early stages of hematopoiesis 1, 2. Using the “difference from normal” flow cytometric approach, low levels of neoplastic leukemia were distinguished from normal regenerating myeloid cells based on the detection of small populations of viable cells exhibiting aberrant antigen expression.…”

Section: Introductionmentioning

confidence: 99%

Consistent quantitative gene product expression: #1. Automated identification of regenerating bone marrow cell populations using support vector machines

Voigt¹,

Brodersen²,

Pardo

et al. 2016

Cytometry Pt A

Self Cite

View full text Add to dashboard Cite

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 Children's 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.

show abstract

Residual disease detected by multidimensional flow cytometry signifies high relapse risk in patients with de novo acute myeloid leukemia: a report from Children's Oncology Group

Cited by 266 publications

References 20 publications

Achieving stringent CR is essential before reduced-intensity conditioning allogeneic hematopoietic cell transplantation in AML

Achieving stringent CR is essential before reduced-intensity conditioning allogeneic hematopoietic cell transplantation in AML

Consistent quantitative gene product expression: #2. Antigen intensities on bone marrow cells are invariant between individuals

Consistent quantitative gene product expression: #1. Automated identification of regenerating bone marrow cell populations using support vector machines

Contact Info

Product

Resources

About