scholarly journals Database-Guided Analysis for Immunophenotypic Diagnosis and Follow-Up of Acute Myeloid Leukemia With Recurrent Genetic Abnormalities

2021 ◽  
Vol 11 ◽  
Author(s):  
Carmen-Mariana Aanei ◽  
Richard Veyrat-Masson ◽  
Cristina Selicean ◽  
Mirela Marian ◽  
Lauren Rigollet ◽  
...  
Keyword(s):  
Myeloid Leukemia ◽  
Residual Disease ◽  
Hematologic Malignancies ◽  
Multicolor Flow Cytometry ◽  
Phenotypic Differences ◽  
Molecular Tests ◽  
Genetic Abnormalities ◽  
Myeloid Leukemias ◽  
Acute Myeloid

Acute myeloid leukemias (AMLs) are hematologic malignancies with varied molecular and immunophenotypic profiles, making them difficult to diagnose and classify. High-dimensional analysis algorithms might increase the utility of multicolor flow cytometry for AML diagnosis and follow-up. The objective of the present study was to assess whether a Compass database-guided analysis can be used to achieve rapid and accurate diagnoses. We conducted this study to determine whether this method could be employed to pilote the genetic and molecular tests and to objectively identify different-from-normal (DfN) patterns to improve measurable residual disease follow-up in AML. Three Compass databases were built using Infinicyt 2.0 software, including normal myeloid-committed hematopoietic precursors (n = 20) and AML blasts harboring the most frequent recurrent genetic abnormalities (n = 50). The diagnostic accuracy of the Compass database-guided analysis was evaluated in a prospective validation study (125 suspected AML patients). This method excluded AML associated with the following genetic abnormalities: t(8;21), t(15;17), inv(16), and KMT2A translocation, with 92% sensitivity [95% confidence interval (CI): 78.6%–98.3%] and a 98.5% negative predictive value (95% CI: 90.6%–99.8%). Our data showed that the Compass database-guided analysis could identify phenotypic differences between AML groups, representing a useful tool for the identification of DfN patterns.

scholarly journals Immunophenotyping Investigation of Minimal Residual Disease Is a Useful Approach for Predicting Relapse in Acute Myeloid Leukemia Patients

Blood ◽  
1997 ◽  
Vol 90 (6) ◽  
pp. 2465-2470 ◽  
Author(s):  
J.F. San Miguel ◽  
A. Martı́nez ◽  
A. Macedo ◽  
M.B. Vidriales ◽  
C. López-Berges ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Multidrug Resistance ◽  
Minimal Residual Disease ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Leukemic Cells ◽  
Rhodamine 123 ◽  
Minimal Residual ◽  
Acute Myeloid

Abstract A high complete remission rate is currently achieved in patients with acute myeloid leukemia (AML). However, many patients eventually relapse due to the persistence of low numbers of residual leukemic cells that are undetectable by conventional cytomorphologic criteria (minimal residual disease [MRD]). Using immunophenotypic multiparametric flow cytometry, we have investigated in sequential studies (diagnosis and follow-up) the impact of MRD detection on the outcome of 53 AML patients that had achieved morphologic remission with standard AML protocols and displayed at diagnosis an aberrant phenotype. Patients were studied at diagnosis with a panel of 35 monoclonal antibodies in triple staining combinations for detection of aberrant or uncommon phenotypic features. According to these features, a patient's probe was custom-built at diagnosis for the identification of possible residual leukemic cells during follow-up. The level of MRD at the end of induction and intensification therapy correlated with the number of relapses and relapse-free survival (RFS). Thus, patients with more than 5 × 10−3 residual cells (5 residual cells among 1,000 normal bone marrow [BM] cells) identified as leukemic by immunophenotyping in the first remission BM showed a significant higher rate of relapse (67% v 20% for patients with less than 5 × 10−3 residual cells; P = .002) and a lower median RFS (17 months v not reached; P = .01). At the end of intensification, with a cut-off value of 2 × 10−3 leukemic cells, AML patients also separated into two distinct groups with relapse rates of 69% versus 32% (P = .02), respectively, and median RFS of 16 months versus not reached (P = .04). In addition, overall survival was also significantly related to the level of residual cells in the marrow obtained at the end of induction and particularly after intensification therapy (P = .008). Furthermore, we have explored whether residual disease was related with the functional expression of multidrug resistance (MDR-1) at diagnosis as assessed by the rhodamine-123 assay. Patients with ≥5 × 10−3 residual leukemic cells at the end of induction therapy had a significantly higher rhodamine-123 efflux (mean, 56% ± 24%) than those with less than 5 × 10−3 residual cells (mean, 32% ± 31%; P = .04). Finally, multivariate analysis showed that the number of residual cells at the end of induction or intensification therapy was the most important prognostic factor for prediction of RFS. Overall, our results show that immunophenotypical investigation of MRD strongly predicts outcome in patients with AML and that the number of residual leukemic cells correlates with multidrug resistance.

scholarly journals Measurable residual disease monitoring in acute myeloid leukemia with t(8;21)(q22;q22.1): results from the AML Study Group

Blood ◽  
2019 ◽  
Vol 134 (19) ◽  
pp. 1608-1618 ◽  
Author(s):  
Frank G. Rücker ◽  
Mridul Agrawal ◽  
Andrea Corbacioglu ◽  
Daniela Weber ◽  
Silke Kapp-Schwoerer ◽  
...  
Keyword(s):  
Myeloid Leukemia ◽  
Residual Disease ◽  
Disease Monitoring ◽  
Study Group ◽  
Relapse Risk ◽  
Time Points ◽  
Key Points ◽  
Measurable Residual Disease ◽  
Acute Myeloid

Key Points MRD assessment in t(8;21) AML allows identification of patients at high relapse risk at defined time points during treatment and follow-up. MRD− after treatment is the most favorable factor for relapse risk and survival, and serial MRD analyses define cutoffs predicting relapse.

Significance of Minimal Residual Disease Monitored by Quantitative Assessment of WT1gene in Patients in First Remission of Acute Myeloid Leukemia Before Allogeneic Stem Cell Transplantation

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4485-4485
Author(s):  
Veronika Válková ◽  
Jaroslav Polak ◽  
Marketa Markova ◽  
Hana Hájková ◽  
Antonin Vitek ◽  
...  
Keyword(s):  
Gene Expression ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Conditioning Regimen ◽  
Wt1 Gene ◽  
Significant Difference ◽  
Induction Failure ◽  
Minimal Residual ◽  
Acute Myeloid

Abstract Abstract 4485 Purpose Thanks to the development of knowledge in the field of molecular biology, the great progress has been done in risk stratification of patients with acute myeloid leukemia (AML) at diagnosis, in recent years. Based on the recommendations of international expert groups there were identified the patients who may benefit from the allogeneic stem cell transplantation (allo-SCT) as a consolidation of first complete remission (CR). In the absence of an universal marker for minimal residual disease (MRD) measurements, there is still little information about the importance of MRD prior to allo-SCT. Our department has a very good experience with quantitative monitoring of WT1 gene expression as a marker of MRD during treatment of AML. The aim was to retrospectively evaluate the significance of MRD in patients indicated for allo-SCT in 1.CR. Patients and methods Overall 35 patients (pts) in the first morphological CR were transplanted from April 2005 - July 2011. Median age was 46 years (range; 20–63), mens 14, women 21, three good risk, intermediate risk 23, high risk 7 (NA 3). A total of 19 pts achieved CR after second induction (salvage), 11 pts were in 1st iCR. Induction 3+7 was given to 31 pts (4x other), as consolidation has been used HIDAC in 28 pts (7x other). As the graft, peripheral blood stem cells were used in 27 pts, bone marrow in 8 pts. The donor was identical sibling in 15 pts (1x mismatched sibling), matched unrelated donor (MUD) in 10 pts and mismatched UD in 9 pts. Conditioning regimen was myeloablative in 29 pts, reduced-intensity in 6 pts. Median follow-up was 18 months (range; 2–56). The expression of WT1 gene was measured by real-time polymerase chain reaction in peripheral blood according to the European Leukemia Net recommendations. The WT1 expression was related to the expression of a reference gene and the results were calculated with a number of WT1 copies related to 104 copies of ABL gene. The upper limit of normal WT1 expression was set as 50 copies of WT1 to 104 copies of ABL. Before allo-SCT, 25 pts were WT1-negative, ten pts were WT1-positive. Results When comparing the two groups according the MRD status, there was not significant difference in terms of age, risk groups, first induction failure, number of iCR, induction or consolidation type. Also, type of graft, conditioning regimen, or HSCT-CI was not significantly different. The group of WT1-positive pts had more unrelated donors, more aGVHD and shorter follow-up. In terms of cGVHD, the groups were comparable. When comparing the overall survival (OS) and cumulative relapse incidence (RI) of the entire group in terms of: risk group, first induction failure, iCR, consolidations number and incidence of aGVHD, we found no significant difference. Pts with cGVHD had a better OS, lower RI with comparable non-relapse mortality (NRM). In contrast, the MRD status measured by WT1 gene expression appears as clearly significant factor. The outcome of WT1-positive pts is significantly worse in terms of OS (55% vs 83% at 3 years, p = 0.03), RI (50% vs 11% at 3 years, p = 0.008), and there is a trend toward higher NRM (23% vs 5% in 3 years, p = 0.08). Conclusion Our results show that MRD status measured by WT1 gene expression in patients with AML in 1.CR significantly affects their future prognosis. Opportunities to influence the unfavorable prognosis of MRD-positive patients may be more intensive pre-transplantation therapy or earlier immunomodulatory intervention after allo-SCT (pre-emptive DLI). The larger prospective studies are necessary to confirm this hypothesis. The study was supported by scientific project MZ 00023736 granted by the Ministry of Health, Czech Republic. Disclosures: No relevant conflicts of interest to declare.

Impact Of Anthracycline Dose Intensification On Minimal Residual Disease and Outcome Of Core Binding Factors Acute Myeloid Leukemias

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2681-2681 ◽  
Author(s):  
Thomas Prebet ◽  
Sarah Bertoli ◽  
Eric Delabesse ◽  
Marie-Joelle Mozziconacci ◽  
Anne Etienne ◽  
...  
Keyword(s):  
Minimal Residual Disease ◽  
Induction Chemotherapy ◽  
Residual Disease ◽  
Lower Probability ◽  
Molecular Response ◽  
Dose Intensification ◽  
Myeloid Leukemias ◽  
Minimal Residual ◽  
Acute Myeloid

Abstract Introduction Acute myeloid leukemias (AML) harboring core binding factor (CBF) alterations (namely t(8;21), inv(16), t(16;16) or variants) are associated with a favorable outcome and are highly sensitive to repeated cycles of high dose cytarabine (HD-Cy, Byrd Blood 2002). Evaluation of minimal residual disease (MRD) by RQ-PCR after first consolidation was recently shown as an important prognosis indicator in CBF AML patients (pts) treated with induction chemotherapy followed by repeated cycles of HD-Cy (Jourdan et al Blood 2013). In the recent years, several studies showed that dose intensification of anthracyclins (Daunomycin 90mg/m2 3 days vs. 45mg/m2) improved survival of younger AML patients (Fernandez NEJM 2009 Lowenberg NEJM 2009). However, there is only limited data for CBF AML. In the present report, we analyzed the MRD levels and the outcome of patients treated either with standard dose or intensified daunomycin (DNR) induction regimens. Patients and Methods This is a retrospective multicenter study. Patients were included consecutively. CBF AML was defined by presence of CBF alteration either by karyotyping, FISH, or RQ-PCR. All pts were treated by induction chemotherapy associating cytarabine 200mg/m2/d day 1-7 CIV with DNR 60mg/m2/d 3 days for patients before 2010 or DNR 90mg/m2/d 3 days between 2010 and 2012. 2 to 3 cycles of HD-Cy were planned for consolidation therapy. For pts after 2005, CBF transcript was assessed by RQ-PCR before induction, after induction (MRD1), cycle 1 (MRD2), and cycle 2 (MRD3) of consolidation. Optimal molecular response was defined as a 1000 fold reduction (3 log) of normalized CBF transcript compared to diagnosis level (Jourdan Blood 2013). Patients with less than 3 log reduction at MRD2 were eligible for allogeneic transplantation. Results 111 patients were evaluated. It included 87 pts treated with DNR60 (37 with molecular follow-up) and 25 pts treated with DNR90 (24 with molecular follow-up). Median age was 42 years for both cohorts. Presence of t(8;21) or inv(16) was detected in 40 and 47 pts of DNR60 cohort, and 6 and 19 pts of DNR90 cohort. Median WBC and platelet counts at diagnosis were 17G/L, 47G/l in the DNR60 cohort and 10G/l, 88G/l in the DNR90 cohort. Median bone marrow blast counts were 70% and 59% respectively. CBF transcript levels at diagnosis were comparable between the 2 groups. All but one patient achieved CR in DNR60 cohort and all patients achieved CR in the DNR90 cohort. 2-years probability of overall survival were 78% and 100% respectively (p=NS). 2-years cumulative incidence of hematologic relapse were 41% and 13% respectively (p=0.04) with 2/3 of the relapses within the first year of follow-up. For patients with molecular follow-up, median CBF transcript level reduction after induction (MRD1) was 2.5 log for DNR60 and 3.7 log for DNR90 (p=0.002). After the first (MRD2) and second cycles (MRD3) of HD-Cy, difference was no longer significant 4.3 log vs. 5 (p=0.12) and 5 log vs.5 log respectively. Percentage of patients achieving optimal molecular response after induction, consolidation 1 and consolidation 2 were respectively: 36% vs. 63% (p=0.02); 75% vs. 91% (p=0.11); 83% vs. 100% (p=0.04). Achievement of molecular response after induction (MRD1) was associated with a lower probability of relapse (18% vs. 45%, p=0.04) whereas no difference was observed when MRD was evaluated after consolidation 1 (MRD2). Of note, OS and RFS of patients treated with DNR60 and with or without molecular follow-up were similar and in line with the recent publication of the French group. Conclusion This retrospective study suggests a benefit of dose-intensification of DNR90 over DNR60 in patients with CBF-AML in terms of early molecular responses and relapse free survival. This shows that molecular remission is a suitable surrogate for RFS in this population. Finally, it provides further evidence that the chemosensitivity of CBF AML is not restricted to HD-Cy. Disclosures: No relevant conflicts of interest to declare.

WT1 Monitoring of Minimal Residual Disease (MRD) in Patients with Acute Myeloid Leukemia

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 3695-3695 ◽  
Author(s):  
Michele Malagola ◽  
Crisitina Skert ◽  
Enrico Morello ◽  
Francesca Antoniazzi ◽  
Erika Borlenghi ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Stem Cell ◽  
Myeloid Leukemia ◽  
Research Funding ◽  
Residual Disease ◽  
Newly Diagnosed ◽  
End Of Treatment ◽  
Minimal Residual ◽  
Acute Myeloid

Abstract Background: Although a complete remission (CR) can be achieved in 70-80% of newly diagnosed acute myeloid leukemia (AML) patients, relapses occur in up to the 50% of cases. Thus, minimal residual disease (MRD) monitoring is a major issue for early detection of patients at high-risk of treatment failure and relapse. Aim: to dynamically evaluate WT1 pan-leukemic molecular marker of MRD in patients with AML. Matherial and methods: 107 newly diagnosed AML patients consecutively treated between 2010 and 2013 were monitored with quantitative WT-1 from bone marrow (BM) and peripheral blood (PB) at baseline, after induction, after the first consolidation course, before allogeneic stem cell transplantation (allo-SCT), at the 3rd and the 6th month after transplantation Results: At diagnosis, 104/107 (97%) had increased PB and BM WT1 levels assessed according to the ELN assay. Eighty-eight out of 107 patients (82%) achieved a complete remission (CR) after induction, 30/88 (34%) relapsed during follow up and 24/107 (22%) were addressed to allogeneic stem cell transplantation (allo-SCT). By univariate analysis, PB-WT > 50x10^4/ABL and BM-WT1 > 250x10^4/ABL after induction (PB: p=0.02; BM: p=0.04), after consolidation (PB: p=0.003), at the end of treatment (PB and BM: p=0.001), at 3rd month of follow up (PB and BM: p=0.005) and at 6th month of follow up (PB: p=0.005) were associated with a reduced overall survival (OS). By multivariate analysis, a BM-WT1 > 250 x 10^4/ABL at the end of treatment was significantly associated with a reduced OS. In order to adapt the cut-off of WT1 in our series of patients, we considered WT1 levels as continuous variables and categorized them at approximately the 25th, 50th, and 75th percentile. A cut-off of PB-WT1 > 25x10^4/ABL and BM-WT1 > 125x10^4/ABL at the end of the treatment program was identified as correlated with reduced leukemia-free survival (LFS) and OS (p=0.001). Similarly, and restricting the analysis on the 24 patients allo-transplanted in CR, 8/11 (73%) with pre-transplant PB-WT1 ≥ 5 and 4/13 (31%) with PB-WT1 < 5 relapsed, respectively (p=0.04). The incidence of relapse was higher in AML patients with PB-WT1 ≥ 5 measured at 3rd (56% vs 38%; p=0.43) and 6th month (71% vs 20%; p=0.03) after allo-SCT. Interestingly, 5/5 (100%) patients with pre-transplant PB-WT1 ≥ 5 who never reduced this level at 3rd or 6th month after allo-SCT experienced a disease recurrence. Conclusions: our data, although retrospectively collected, show that WT1 monitoring may be useful to predict the relapse in AML patients. Acknowledgments: This work was supported in part by Banca di Credito Cooperativo di Pompiano e Franciacorta and Lions Club Bassa Bresciana Association. Disclosures Russo: Celgene: Research Funding; Gilead: Research Funding; Novartis: Consultancy.

Activity of venetoclax-based therapy in TP53-mutated acute myeloid leukemia.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. 7034-7034
Author(s):  
Mahran Shoukier ◽  
Marina Konopleva ◽  
Courtney Denton Dinardo ◽  
Farhad Ravandi ◽  
Michael Andreeff ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Patient Characteristics ◽  
Cytogenetic Response ◽  
Effective Treatment Option ◽  
Whole Exome ◽  
Poor Outcomes ◽  
Acute Myeloid

7034 Background: Mutations in TP53 are associated with low response rates to standard therapy and poor outcomes in patients (pts) with acute myeloid leukemia (AML). Combination therapy with the BCL2 inhibitor venetoclax (VEN) has emerged as an effective treatment option for pts with AML. Methods: We reviewed pts with TP53-mutated AML treated with VEN-based therapy between 2014-2018. Mutation testing was performed using a whole-exome next-generation sequencing panel. We analyzed the characteristics of these pts, responses to therapy, and outcomes. Results: Sixty nine pts with TP53-mutated AML treated with VEN were identified, 36 (52%) in frontline & 33 (48%) in the salvage (R/R) setting (Table). The median follow up was 4.5 [0.5 - 48.5] and 8 [1 - 46.5] months for frontline & R/R pts, respectively. Karyotype was complex in 32 (88%) and 29 (88%) pts in the frontline & R/R cohorts, respectively. In the R/R cohort, the number of median prior treatments was 2 [0 – 8]. VEN was given in combination with: 1) Hypomethylating agents (HMA) (87%), 2) FLAG-Ida (3%), 3) Low dose Ara-C (4%), or 4) CPX-351 (6%). The overall response rate (ORR) was 47% & 24% in frontline and R/R pts, respectively. All 6 pts with negative minimal residual disease (MRD) achieved complete cytogenetic response after taking VEN % remain in complete remission (CR) with a median of 3.4 [1.7-4.7] months. Two pts (both R/R) underwent allogeneic stem cell transplantation. Conclusions: VEN based therapy was associated with similar ORR, but higher CR rates in TP53 mutated AML compared with HMAs alone. Larger studies with longer follow up are needed to determine the role of VEN-based therapy in this difficult subset. Patient characteristics and outcome. [Table: see text]

scholarly journals Prognostic Value of Minimal Residual Disease before Allogeneic Hematopoietic Stem Cell Transplantation in Patients with Acute Myeloid Leukemia

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 5733-5733
Author(s):  
Olga Pérez-López ◽  
Teresa Caballero-Velázquez ◽  
Enrique Colado ◽  
Sara Alonso ◽  
José González-Campos ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Prognostic Value ◽  
Residual Disease ◽  
Hematopoietic Stem ◽  
Multiparametric Flow Cytometry ◽  
Minimal Residual ◽  
Acute Myeloid

Abstract Introduction Several studies have shown that the minimal residual disease (MRD) in acute myeloid leukemia (AML) patients has a prognostic value after induction and consolidation therapy. Nevertheless the relapse is the most important cause of treatment failure in these patients, although they achieved a negative MRD, and even after an allogeneic hematopoietic stem cell transplantation (allo-HSCT). Nowadays, the value of the MRD before allogeneic BMT is still controversial. Method Multicentric study where we have studied correlative AML patients who went under an allo-HSCT in a situation of complete response, between 2012 and April'18. The MRD was analyzed by 8-coloured multiparametric flow cytometry, at least with 2 tubes per patient and 1,000,000 events per tube. We evaluated the prognostic value of the MRD before allo-HSCT. Results Between January'12 and April'18 we have gathered 90 allogeneic BMT in AML patients who were in CR, with a median age of 45 years old (17 - 66). The pre-HSCT situation was 1st complete remission (CR) in 75 patients and 2nd CR in 15. In 45 patients the conditioning regimen was myeoablative. In the group of patients (67) where we could know the risk group at diagnosis, the distribution was: low risk 18%, intermediate risk 59.7% and high risk 22.4%. The 46.7% of the donors were not related. In the last follow-up after allo-HSCT 24 patients have suffered a relapse (26.7%) and 41 (45.5%) have died (17 cases of mortality related to the transplant and 24 not related). In the global analysis the median follow-up of the overall survival (OS) was 37.5 months. Among the 90 patients, MRD was valuable in 86. Ten of 59 patients (16.9%) with negative MRD relapsed vs 12/27 (44.4%) with positive MRD, p= 0.016. If we consider only patients in 1st CR, 9/50 (18%) patients with negative MRD relapsed vs 10/22 (45.5%) with positive MRD, p= 0.02. This statistically significant difference does not exist if we consider only patients in 2nd CR. The median follow-up of OS and event free survival (EFS) was not reached in the negative MRD group and 571 days and 299 days in the positive MRD group. OS and EFS at 2 years after transplantation were 65% and 64% in the negative MRD group and 42% and 37% in the positive MRD group, p= 0.03 and p= 0.008 respectively (figure 1). Conclusions The detected MRD by 8-colour multiparametric flow cytometry previous an allo-HSCT in patients with AML in 1st CR is a prognostic factor in terms of relapse. Patients with a positive MRD before the allo-HSCT have a poorer OS and EFS than the patients with a negative MRD. Figure 1. Figure 1. Disclosures No relevant conflicts of interest to declare.

scholarly journals Immunophenotyping Investigation of Minimal Residual Disease Is a Useful Approach for Predicting Relapse in Acute Myeloid Leukemia Patients

Blood ◽  
1997 ◽  
Vol 90 (6) ◽  
pp. 2465-2470 ◽  
Author(s):  
J.F. San Miguel ◽  
A. Martı́nez ◽  
A. Macedo ◽  
M.B. Vidriales ◽  
C. López-Berges ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Multidrug Resistance ◽  
Minimal Residual Disease ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Leukemic Cells ◽  
Rhodamine 123 ◽  
Minimal Residual ◽  
Acute Myeloid

A high complete remission rate is currently achieved in patients with acute myeloid leukemia (AML). However, many patients eventually relapse due to the persistence of low numbers of residual leukemic cells that are undetectable by conventional cytomorphologic criteria (minimal residual disease [MRD]). Using immunophenotypic multiparametric flow cytometry, we have investigated in sequential studies (diagnosis and follow-up) the impact of MRD detection on the outcome of 53 AML patients that had achieved morphologic remission with standard AML protocols and displayed at diagnosis an aberrant phenotype. Patients were studied at diagnosis with a panel of 35 monoclonal antibodies in triple staining combinations for detection of aberrant or uncommon phenotypic features. According to these features, a patient's probe was custom-built at diagnosis for the identification of possible residual leukemic cells during follow-up. The level of MRD at the end of induction and intensification therapy correlated with the number of relapses and relapse-free survival (RFS). Thus, patients with more than 5 × 10−3 residual cells (5 residual cells among 1,000 normal bone marrow [BM] cells) identified as leukemic by immunophenotyping in the first remission BM showed a significant higher rate of relapse (67% v 20% for patients with less than 5 × 10−3 residual cells; P = .002) and a lower median RFS (17 months v not reached; P = .01). At the end of intensification, with a cut-off value of 2 × 10−3 leukemic cells, AML patients also separated into two distinct groups with relapse rates of 69% versus 32% (P = .02), respectively, and median RFS of 16 months versus not reached (P = .04). In addition, overall survival was also significantly related to the level of residual cells in the marrow obtained at the end of induction and particularly after intensification therapy (P = .008). Furthermore, we have explored whether residual disease was related with the functional expression of multidrug resistance (MDR-1) at diagnosis as assessed by the rhodamine-123 assay. Patients with ≥5 × 10−3 residual leukemic cells at the end of induction therapy had a significantly higher rhodamine-123 efflux (mean, 56% ± 24%) than those with less than 5 × 10−3 residual cells (mean, 32% ± 31%; P = .04). Finally, multivariate analysis showed that the number of residual cells at the end of induction or intensification therapy was the most important prognostic factor for prediction of RFS. Overall, our results show that immunophenotypical investigation of MRD strongly predicts outcome in patients with AML and that the number of residual leukemic cells correlates with multidrug resistance.

Identification of a Set of Seven Genes for the Monitoring of Minimal Residual Disease in Pediatric Acute Myeloid Leukemia.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1461-1461
Author(s):  
Daniel Steinbach ◽  
Alexander Schramm ◽  
Angelika Eggert ◽  
Susann Wittig ◽  
Nadine Pfaffendorf ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Minimal Residual Disease ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Wilms Tumor ◽  
Leukemic Cells ◽  
Genome Wide ◽  
Minimal Residual ◽  
Acute Myeloid

Abstract A stepwise approach which combined genome wide expression profiling and a TaqMan realtime PCR based screening was used to identify new markers for the monitoring of minimal residual disease (MRD) in acute myeloid leukemia (AML). Leukemic cells from 52 children with AML were analyzed. Seven genes were identified which are vastly over-expressed in many patients with AML compared to healthy bone marrow: CCL23, GAGED2, MSLN, SPAG6, and ST18 as well as the previously described markers WT1 and PRAME. This set of genes was analyzed in 141 follow-up samples from 25 patients. The expression of all genes decreased to normal levels in patients who achieved a continuous complete remission. Elevated levels of MRD markers were found prior to relapse in 7 out of 10 patients who relapsed. This set of genes should allow a sensitive and specific monitoring of MRD in AML. Notably, some of these markers could also serve as therapeutic targets or might be involved in leukemogenesis. MSLN is already used as a target for immunotherapy in clinical trials in other malignancies. GAGED2 and SPAG6 belong to the family of cancer testis genes which are also studied intensively as targets for immunotherapy. ST18 is a recently discovered tumor suppressor which was not yet described in hematological malignancies. CCL23 is a chemokine that inhibits the proliferation of healthy hematological stem cells. Names, symbols, and geneID of seven MRD markers Gene Symbol Gene Name GeneID CCL23 chemokine (C–C motif) ligand 23 6368 GAGED2 G antigen, family D, 2 9503 MSLN Mesothelin 10232 SPAG6 sperm associated antigen 6 9576 ST18 suppression of tumorigenicity 18 9705 WT1 Wilms tumor 1 7490 PRAME preferentially expressed antigen in melanoma 23532

Simple, Gel-Based Determination of Nucleophosmin Mutations in Acute Myeloid Leukemias.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4268-4268
Author(s):  
Kim P. Ahrens
Keyword(s):  
Myeloid Leukemia ◽  
Residual Disease ◽  
Size Difference ◽  
Myeloid Leukemias ◽  
The Individual ◽  
Mutant Genes ◽  
Small Product ◽  
Minimal Residual ◽  
Acute Myeloid

Abstract Mutations in the Nucleophosmin, member 1 (NPM1) gene, have been shown to be significant for prognosis and treatment of cytogenetically normal (CN) patients with acute myeloid leukemia (AML). To increase sensitivity for minimal residual disease (MRD) detection, especially in AMLs without a distinct phenotype or genetic marker, some assays have specifically targeted the individual mutation, which has led to an increased complexity of these tests. Other assays use expensive equipment such as capillary electrophoresis to differentiate between the usual 4bp size difference in the mutated gene and the wildtype(wt) gene. We developed a simple PCR-based procedure that amplifies the exon 12 region of the NPM1 gene and produces a small product (156bp for the wt and 160 for the mutant genes) using RNA that can be separated on a 4% metaphor agarose gel. This approach is clinically relevant since it can easily distinguish greater than 95% of the mutations. Eight of 24 AMLs of various types were positive for the mutation using our technique and those that were cloned and sequenced yielded the most common insertion TCTG, confirming its accuracy.

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