Multiparameter Flow Cytometry in the Diagnosis and Management of Acute Leukemia

2011 ◽  
Vol 135 (1) ◽  
pp. 44-54 ◽  
Author(s):  
John M. Peters ◽  
M. Qasim Ansari
Keyword(s):  
Flow Cytometry ◽  
Acute Leukemia ◽  
Minimal Residual Disease ◽  
Residual Disease ◽  
Accurate Diagnosis ◽  
Response To Therapy ◽  
Multiparameter Flow Cytometry ◽  
Acute Leukemias ◽  
Diagnosis And Management ◽  
Minimal Residual

Abstract Context—Timely and accurate diagnosis of hematologic malignancies is crucial to appropriate clinical management. Acute leukemias are a diverse group of malignancies with a range of clinical presentations, prognoses, and preferred treatment protocols. Historical classification systems relied predominantly on morphologic and cytochemical features, but currently, immunophenotypic, cytogenetic, and molecular data are incorporated to define clinically relevant diagnostic categories. Multiparameter flow cytometry provides rapid and detailed determination of antigen expression profiles in acute leukemias which, in conjunction with morphologic assessment, often suggests a definitive diagnosis or a narrow differential. Many recurrent molecular or cytogenetic aberrations are associated with distinct immunophenotypic features, and therefore flow cytometry is an important tool to direct further testing. In addition, detection of specific antigens may have prognostic or therapeutic implications even within a single acute leukemia subtype. After initial diagnosis, a leukemia's immunophenotypic fingerprint provides a useful reference to monitor response to therapy, minimal residual disease, and recurrence. Objective—To provide an overview of the application of flow cytometric immunophenotyping to the diagnosis and management of acute leukemias, including salient features of those entities described in the 2008 World Health Organization classification. Data Sources—Published articles pertaining to flow cytometry, acute leukemia classification, and experiences of a reference flow cytometry laboratory. Conclusion—Immunophenotypic evaluation is essential to accurate diagnosis and classification of acute leukemia. Multiparameter flow cytometry provides a rapid and effective means to collect this information, as well as providing prognostic information and a modality for minimal residual disease evaluation.

scholarly journals Unsupervised Flow Cytometry Analysis Allows for an Accurate Identification of Minimal Residual Disease Assessment in Acute Myeloid Leukemia

Cancers ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 629
Author(s):  
Jean Philippe Vial ◽  
Nicolas Lechevalier ◽  
Francis Lacombe ◽  
Pierre-Yves Dumas ◽  
Audrey Bidet ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Minimal Residual Disease ◽  
Residual Disease ◽  
Response To Therapy ◽  
Disease Assessment ◽  
Multiparameter Flow Cytometry ◽  
Accurate Identification ◽  
Self Organizing Maps ◽  
Minimal Residual

The assessment of minimal residual disease (MRD) is increasingly considered to monitor response to therapy in hematological malignancies. In acute myeloblastic leukemia (AML), molecular MRD (mMRD) is possible for about half the patients while multiparameter flow cytometry (MFC) is more broadly available. However, MFC analysis strategies are highly operator-dependent. Recently, new tools have been designed for unsupervised MFC analysis, segregating cell-clusters with the same immunophenotypic characteristics. Here, the Flow-Self-Organizing-Maps (FlowSOM) tool was applied to assess MFC-MRD in 96 bone marrow (BM) follow-up (FU) time-points from 40 AML patients with available mMRD. A reference FlowSOM display was built from 19 healthy/normal BM samples (NBM), then simultaneously compared to the patient’s diagnosis and FU samples at each time-point. MRD clusters were characterized individually in terms of cell numbers and immunophenotype. This strategy disclosed subclones with varying immunophenotype within single diagnosis and FU samples including populations absent from NBM. Detectable MRD was as low as 0.09% in MFC and 0.051% for mMRD. The concordance between mMRD and MFC-MRD was 80.2%. MFC yielded 85% specificity and 69% sensitivity compared to mMRD. Unsupervised MFC is shown here to allow for an easy and robust assessment of MRD, applicable also to AML patients without molecular markers.

Disease Response Guiding Therapy in Acute Lymphocytic Leukemia: Pivotal Role of Minimal Residual Disease

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. SCI-34-SCI-34
Author(s):  
Michael A. Pulsipher
Keyword(s):  
Flow Cytometry ◽  
High Risk ◽  
Minimal Residual Disease ◽  
Lower Risk ◽  
Residual Disease ◽  
Response To Therapy ◽  
Risk Patients ◽  
Very High ◽  
Minimal Residual ◽  
Risk Of Relapse

In spite of an explosion of data regarding mutations associated with childhood ALL, to date these key genetic changes rarely have been the driver of therapy. Clinical parameters at presentation (WBC, age, T- vs. B-lineage, etc.) have dictated initial risk stratification and induction approaches, followed by risk-adapted therapy based upon leukemic response measured by minimal residual disease (MRD, either PCR- or flow cytometry-based). With minor variations, rapid disappearance of peripheral MRD, followed by significant clearance from the marrow after induction, and most importantly, the level of MRD after consolidation have allowed clear distinctions in outcomes that have driven intensification or de-intensification of therapy resulting in improved outcomes. Although specific gene mutations have been associated with risk, MRD has further identified better risk patients within genetic subgroups. For patients noted to be very high risk who are candidates for hematopoietic cell transplantation (HCT), the presence of MRD both pre- and post-transplant has been associated with increased risk of relapse; the risk being modified by level of MRD, whether or not GVHD occurs after HCT, and timing after HCT when MRD is measured. In lower risk patients being treated with chemotherapy and higher risk patients eligible for HCT, more sensitive approaches to flow cytometry and PCR, as well as next-generation sequencing (NGS) MRD approaches (sensitive to 1/10^7 cells) are currently being tested. It is not clear yet whether NGS-MRD offers substantial improvements in patients treated with chemotherapy, as broad-based testing is underway; the latest comparative outcomes will be presented. There is evidence of a striking improvement in our ability to define patients who will do very will after transplant (not relapse), and preliminary evidence that post-HCT NGS MRD testing is more sensitive that other methodologies in defining risk of relapse after transplant. As the latest information about the ability of different approaches to MRD is shown in this session, we will also present how response to therapy based upon MRD interacts with various genetic subtypes (Ph+ ALL, extreme hypodiploidy, etc.). Even in subclasses that are considered very high risk based solely upon genetics, measurement of MRD can define higher and lower risk groups. Going forward, as more and different types of patients are subcategorized and treated with targeted agents based upon specific mutations, it is likely MRD response will continue to be important in mapping intensity of approach and defining children at highest risk of relapse who might benefit from HCT or other cellular therapeutic approaches. Disclosures Pulsipher: Novartis: Membership on an entity's Board of Directors or advisory committees.

Minimal Residual Disease Assessed by Multiparameter Flow Cytometry in Multiple Myeloma: Impact on Outcome in the Medical Research Council Myeloma IX Study

2013 ◽  
Vol 31 (20) ◽  
pp. 2540-2547 ◽  
Author(s):  
Andy C. Rawstron ◽  
J. Anthony Child ◽  
Ruth M. de Tute ◽  
Faith E. Davies ◽  
Walter M. Gregory ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Flow Cytometry ◽  
Medical Research ◽  
Minimal Residual Disease ◽  
Induction Therapy ◽  
Research Council ◽  
Medical Research Council ◽  
Residual Disease ◽  
Multiparameter Flow Cytometry ◽  
Minimal Residual

Purpose To investigate the prognostic value of minimal residual disease (MRD) assessment in patients with multiple myeloma treated in the MRC (Medical Research Council) Myeloma IX trial. Patients and Methods Multiparameter flow cytometry (MFC) was used to assess MRD after induction therapy (n = 378) and at day 100 after autologous stem-cell transplantation (ASCT; n = 397) in intensive-pathway patients and at the end of induction therapy in non–intensive-pathway patients (n = 245). Results In intensive-pathway patients, absence of MRD at day 100 after ASCT was highly predictive of a favorable outcome (PFS, P < .001; OS, P = .0183). This outcome advantage was demonstrable in patients with favorable and adverse cytogenetics (PFS, P = .014 and P < .001, respectively) and in patients achieving immunofixation-negative complete response (CR; PFS, P = .0068). The effect of maintenance thalidomide was assessed, with the shortest PFS demonstrable in those MRD-positive patients who did not receive maintenance and longest in those who were MRD negative and did receive thalidomide (P < .001). Further analysis demonstrated that 28% of MRD-positive patients who received maintenance thalidomide became MRD negative. MRD assessment after induction therapy in the non–intensive-pathway patients did not seem to be predictive of outcome (PFS, P = .1). Conclusion MRD assessment by MFC was predictive of overall outcome in patients with myeloma undergoing ASCT. This predictive value was seen in patients achieving conventional CR as well as patients with favorable and adverse cytogenetics. The effects of maintenance strategies can also be evaluated, and our data suggest that maintenance thalidomide can eradicate MRD in some patients.

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