Functional characterization of minimal residual disease for P-glycoprotein and multidrug resistance protein activity in acute myeloid leukemia

Pol, MA van der; Pater, JM; Feller, N; Westra, AH; Stijn, Amber van; Ossenkoppele, GJ; Broxterman, H. J.; Schuurhuis, G.J.

doi:10.1038/sj.leu.2402245

Cited by 27 publications

(34 citation statements)

References 40 publications

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“…The treatment scheme is shown in Figure 1a. In the HOVON 29 protocol, the first cycle included a standard dose of cytarabine (200 mg/m 2 /day, days 1-7) and idarubicin (12 mg/m 2 /day, days [6][7][8] and in the second cycle, an intermediate dose of cytarabine (1000 mg/m 2 /day, days 1-6) and amsacrine (120 mg/m 2 /day, days [4][5][6]. In the HOVON 42 protocol patients were first randomized to either chemotherapy identical to HOVON 29 or to an intermediate dose of cytarabine (1000 mg/m 2 day, days 1-7) plus idarubicin (12 mg/m 2 /day, days 1-3) followed by a high-dose cytarabine (2000 mg/m 2 /day, days 1, 3, 5 and 7) plus amsacrine (120 mg/m 2 /day, days 2, 4 and 6).…”

Section: Patient Treatmentmentioning

confidence: 99%

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MRD parameters using immunophenotypic detection methods are highly reliable in predicting survival in acute myeloid leukaemia

et al. 2004

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Outgrowth of minimal residual disease (MRD) in acute myeloid leukaemia (AML) is responsible for the occurrence of relapses. MRD can be quantified by immunophenotyping on a flow cytometer using the expression of leukaemia-associated phenotypes. MRD was monitored in follow-up samples taken from bone marrow (BM) of 72 patients after three different cycles of chemotherapy and from autologous peripheral blood stem cell (PBSC) products. The MRD% in BM after the first cycle (n ¼ 51), second cycle (n ¼ 52) and third cycle (n ¼ 30), as well as in PBSC products (n ¼ 39) strongly correlated with relapse-free survival. At a cutoff level of 1% after the first cycle and median cutoff levels of 0.14% after the second, 0.11% after the third cycle and 0.13% for PBSC products, the relative risk of relapse was a factor 6.1, 3.4, 7.2 and 5.7, respectively, higher for patients in the high MRD group. Also, absolute MRD cell number/ml was highly predictive of the clinical outcome. After the treatment has ended, an increase of MRD% predicted forthcoming relapses, with MRD assessment intervals of p3 months. In conclusion, MRD parameter assessment at different stages of disease is highly reliable in predicting survival and forthcoming relapses in AML. Leukemia (2004) IntroductionMinimal residual disease (MRD) cells, present in the bone marrow (BM) of patients with acute myeloid leukaemia (AML) after chemotherapy, are thought to be responsible for the emergence of a relapse. Monitoring of MRD cells by molecular biologic or immunophenotypic detection methods are thought to be of crucial value for defining individualized treatment of patients. Based on MRD frequencies patient-tailored postremission chemotherapy could be designed, which might reduce either the risk of relapse or, in case of withholding therapy to the patient, diminish morbidity and mortality. Also, when impending relapses can be predicted, these might be prevented by early therapeutic interventions. Furthermore, MRD frequency assessment can be used to quantify the leukaemia cell contamination of an autologous peripheral blood stem cell (PBSC) product in order to guide the decision of whether or not to purge, and in the latter case to establish the efficacy of leukaemia cell eradication.Immunophenotypic detection of MRD by flow cytometry can be performed by defining aberrant marker expressions, the socalled leukaemia-associated phenotypes (LAPs), on malignant cells at diagnosis. 1-6 These LAPs are not, or only in very low frequencies, present on normal blood or BM cells. This technique can be successfully applied in up to 80% of the AML patients in contrast to molecular biological techniques, which were until now applicable only in 20-30% of the cases. 7-9 So far, in AML the predictive value of MRD frequency detection using immunophenotyping has been reported by a few groups only. 6,[10][11][12] In adult AML patients, San Miguel et al 6,10 demonstrated the prognostic value of MRD after induction and intensification therapy, while Venditti et al 11 have found this after con...

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Section: Patient Treatmentmentioning

confidence: 99%

“…[1][2][3][4][5][6] These LAPs are not, or only in very low frequencies, present on normal blood or BM cells. This technique can be successfully applied in up to 80% of the AML patients in contrast to molecular biological techniques, which were until now applicable only in 20-30% of the cases.…”

Section: Introductionmentioning

confidence: 99%

MRD parameters using immunophenotypic detection methods are highly reliable in predicting survival in acute myeloid leukaemia

et al. 2004

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“…Extensive validation of the use of cryopreserved material showed no selective loss of subgroups of cells. 6 About 0.5-1 x 10 6 cells were incubated with the MoAbs for 15 min at room temperature, lysed, washed twice and measured by flow cytometry. Data analyses were performed on blasts with dim CD45 staining and low SSC properties 14 and corresponding isotype controls were included for every LAP.…”

Section: Detection and Quantification Of Mrd Frequencies In Pbsc Prodmentioning

confidence: 99%

“…[2][3][4] These studies were performed using combinations of cell surface markers present on leukemia cells, but not or in very low frequencies on normal cells, which allows MRD detection in up to 80% of the patients. [5][6][7] In studying the contamination of MRD cells in PBSC products with these leukemia associated phenotypes (LAPs) using flow cytometry, we found that a high percentage of CD34-positive cells in PBSC products was associated with a high relapse rate. This obser- vation shed another light on the general view that high CD34-positive cell doses are, in terms of hematological recovery, beneficial for patients treated with high-dose chemotherapy followed by PBSC transplantation.…”

Section: Introductionmentioning

confidence: 99%

High percentage of CD34-positive cells in autologous AML peripheral blood stem cell products reflects inadequate in vivo purging and low chemotherapeutic toxicity in a subgroup of patients with poor clinical outcome

et al. 2003

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In this study, a high CD34% in autologous peripheral blood stem cell (PBSC) products from 71 AML patients was associated directly with a high relapse rate (P = 0.006) and inversely with disease-free survival (P = 0.003), irrespective whether patients were transplanted or not. The relapse rate at 12 months was 67% in a group with Ͼ0.8% CD34 + cells and 34% in a group with р0.8% CD34 + cells. Although the percentage of malignant CD34 + cells in the CD34 + compartment in the relapses of the first group was not high (median 8%), the total number of malignant cells as a percentage of WBC was about 13 times higher than for the patients remaining Ͼ12 months in remission. When all patients evaluable were taken together, this frequency of malignant cells correlated strongly with disease-free survival (P Ͻ 0.001). Both this massive mobilization of normal CD34 + cells and high frequency of malignant cells in the subgroup of patients with CD34 Ͼ0.8% and relapse within 12 months indicate an insufficient in vivo purging, as well as low chemotherapeutic bone marrow toxicity. This was confirmed by an inverse correlation between hypoplasia period after the induction therapy and CD34% in PBSC products (P Ͻ 0.002). It is concluded that a subgroup of patients has been identified that might benefit from a more intensive chemotherapeutic treatment.

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“…With the conventional Western blot method, this would not be possible because of the limited number of MRD cells available. We have recently developed a technique that allows the detection of ABC transporter activity in MRD (van der Pol et al, 21 Haematologica, 2003, 88, in press). The present approach was aimed to characterize the role of apoptosisrelated proteins in MRD in AML.…”

Section: Introductionmentioning

confidence: 99%

A flow cytometric method to detect apoptosis-related protein expression in minimal residual disease in acute myeloid leukemia

et al. 2003

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Minimal residual disease (MRD) cells are thought to be responsible for the persistence and relapse of acute myeloid leukemia (AML). Flow cytometric MRD detection by the establishment of a leukemia-associated phenotype (LAP) at diagnosis can be used in 80% of AML patients, allowing detection and functional characterization of MRD in follow-up bone marrow. One of the mechanisms contributing to inefficient chemotherapy is apoptosis resistance. Measuring apoptosis parameters in MRD cells will help to unravel the importance of apoptosis resistance in AML. We therefore developed a fourcolor flow cytometry method that enables establishment of apoptosis-related protein expression such as Bcl-2, Bcl-x L , Mcl-1 and Bax at diagnosis and in MRD. Firstly, validation of this assay using Western blot analysis in five leukemia cell lines showed a significant correlation (R = 0.70: Po0.0001). Secondly, the influence of the permeabilization procedure on LAP expression was investigated in 38 AML samples at diagnosis and in 42 MRD samples. Quantification of the frequency of LAP+ cells with and without permeabilization showed no significant differences (diagnosis: P = 0.57, follow-up: P = 0.43). The flow cytometric protocol thus enables analysis of apoptosis-related proteins at different stages of the disease, which will lead to a better understanding of the role of apoptosis resistance in the emergence of MRD in AML.

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Functional characterization of minimal residual disease for P-glycoprotein and multidrug resistance protein activity in acute myeloid leukemia

Cited by 27 publications

References 40 publications

MRD parameters using immunophenotypic detection methods are highly reliable in predicting survival in acute myeloid leukaemia

MRD parameters using immunophenotypic detection methods are highly reliable in predicting survival in acute myeloid leukaemia

High percentage of CD34-positive cells in autologous AML peripheral blood stem cell products reflects inadequate in vivo purging and low chemotherapeutic toxicity in a subgroup of patients with poor clinical outcome

A flow cytometric method to detect apoptosis-related protein expression in minimal residual disease in acute myeloid leukemia

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