ABC Proteins Activity Remains An Independent Prognostic Factor In 206 AML When Compared to Other Molecular Markers, FLT3, NPM1, CEBPα, and BAALC

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1698-1698
Author(s):  
Pierre Hirsch ◽  
Ruoping Tang ◽  
Christophe Marzac ◽  
Jean-Yves Perrot ◽  
Fanny Fava ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Conflicts Of Interest ◽  
Normal Karyotype ◽  
Functional Assay ◽  
Prognosis Factor ◽  
Prognosis Factors ◽  
Abc Proteins ◽  
Npm1 Mutation ◽  
Molecular Alterations ◽  
Mutational Status

Abstract Abstract 1698 Background: Many clinical and biological features are commonly used in order to predict the probability of response to standard treatment in adult acute myeloid leukaemia (AML). Although cytogenetic analysis provides the major information for prognosis, many molecular alterations like FLT3/ITD, mutations of NPM1, mutations of CEBPα, or BAALC over expression have been described these last years, in order to guide therapeutic choices, especially in patients with normal karyotype. In the other hand, prognostic implications of ABC proteins' expression and functionality, like ABCB1 (Pgp) and others, have been known for years but relationships between ABC proteins and those molecular markers haven't been fully studied. Material and methods: In this retrospective study, we explore the relationships between ABC proteins and these molecular markers, and evaluate whether ABC proteins' activity is an independent prognosis factor in 206 AML, homogenously treated in EORTC protocols. ABC proteins' activity has been assessed with JC1 functional test performed at the time of diagnosis, and all patients were screened from frozen bone marrow or peripheral blood samples for FLT3/ITD, NPM1 mutations, CEBPα mutations, and expression of BAALC. 206 patients aged from 16 to 81 years and treated either in EORTC AML 10, AML 12 or AML 13 from 1996 to 2008 have been included, with a median follow up time for alive patients of five years. Results: In the whole population, 42 patients (20%) showed a high ABC functional assay (JC1+). High ABC activity was associated with NPM1 wild type (p=0.03) and higher BAALC expression (p=0.007). FLT3-ITD was detected in only 2 patients with high ABC functional assay. There was no relationship between high ABC functionality and CEBPα mutational status. In multivariate analysis including age, leukocyte count, cytogenetic, existence of a pre leukemic phase, NPM1, FLT3/ITD, CEBPα and BAALC status and JC1 assay, high ABC proteins' activity was an independent prognosis factor for OS in the whole population (p=0.03). Others independent prognosis factors for OS were age (p=0.0004), cytogenetic (p=0,045), FLT3/ITD (p=0.0096), and NPM1 mutation (p= 0.07). When interesting to normal cytogenetic population (112 patients), high ABC proteins' activity was an independent prognosis factor for DFS (p=0.0107) and OS (p=0.0038). Age (p=0.0012), and FLT3/ITD (p=0.0173) were the only other prognosis factors for OS. In patients with normal cytogenetic, and both NPM1 WT and no FLT3/ITD, only age (p=0.0008) and high ABC proteins' activity (p=0.004) were independent prognosis factors for OS. Conclusion: ABC proteins' activity remains an independent prognosis factor in adult AML, when compared to these molecular factors. Because of its relatively high frequency (around 20 % of patients), and the easiness to perform this test, we think JC1 probe should be used in every AML patients in order to refine the treatment strategy at the time of diagnosis. Disclosures: No relevant conflicts of interest to declare.

Prognostic Impact of Clinical and Molecular Markers in Normal Karyotype AML-Results from the AMLCG 2000 Trial.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3482-3482
Author(s):  
Friederike Schneider ◽  
Eva Hoster ◽  
Marietta Rottenkolber ◽  
Stephanie Schneider ◽  
Annika Dufour ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Risk Stratification ◽  
Multivariate Analyses ◽  
Normal Karyotype ◽  
Molecular Techniques ◽  
Prognostic Impact ◽  
Clinical Parameters ◽  
Single Mutation ◽  
Free Survival ◽  
Molecular Alterations

Abstract Background: Prognosis of AML is influenced by different clinical and molecular alterations. We performed a multivariate analysis including five molecular markers NPM1, FLT-ITD, CEBPA, FLT-TKD and MLL-PTD combined with clinical parameters at initial diagnosis to refine risk stratification. Patients and methods: Prognostic impact of clinical and molecular parameters in respect to OS, EFS, RFS and CR was assessed in 803 patients with normal karyotype included in the AMLCG (German AML Cooperative Group) 2000 trial until 01/2006. Patients were randomly assigned to treatment with TAD (thioguanine, conventional-dose AraC, daunorubicin) followed by HAM (high-dose AraC, mitoxantrone) or with the double-induction regimen consisting of two courses of HAM (quotation Buechner JCO 2006). Patient age ranged from 17 to 85 years (median: 60 yrs). 51% of patients were male, 49% female. 81% of patients had de novo AML. Performance status was normal or slightly impaired in the majority of patients (71% ECOG 0/1). Median blood counts at diagnosis were: Hb: 9.2 g/dl (4.2–16.4 g/dl); WBC: 16.0 G/l (0.1–798.2 G/l); platelets: 58 G/l (0.02–643 G/l), LDH: 410 U/l (8–14332 U/l) and bone marrow (BM) blasts: 80% (6–100%). Molecular markers’ mutation status and all mentioned clinical parameters were included in univariate analyses. In multivariate analyses only univariate significant parameters were used. Results: In 560 patients with all five molecular markers analyzed by routine molecular techniques at diagnosis the frequency of mutations were the following: 52.7% NPM1+, 29.3% FLT3-ITD+, 6.1% FLT3-TKD+, 7.5% MLL-PTD+ and 7.5% CEBPA+. The majority of analyzed patients (44.1%) showed one single mutation only. About one quarter of patients displayed either none (27.5%) or two (26.2%) mutations. A minority of 2.1% had 3 mutations, whereas the combination of four or all five molecular alterations was not found. The most frequent single mutation was NPM1 (28.4%), followed by FLT3-ITD (5.4%), CEBPA (4.8%), MLL-PTD (4.6%) and FLT3-TKD (0.9%). The combination of FLT3-ITD and NPM1 was detected in 18.8% of patients. Complete remission (CR) rate was 65.1%. Median overall survival (OS), event-free survival (EFS) and relapse-free survival (RFS) were 19.3, 7.7 and 17.2 months. Multivariate analyses identified the following parameters to have significant impact on prognosis. OS: NPM1, FLT3-ITD, WBC, age (p<0.0001 each) and CEBPA (p=0.003); EFS/RFS: NPM1, FLT3-ITD and age (p<0.0001 each / p<0.0001 each) and LDH (p=0.020 / p=0.040); CR: NPM1 and age (p=0.001 each). Conclusions: Our data show in a large cohort of 560 patients that at least one molecular marker can be identified in 72.5% of patients with NK-AML. The NPM1 mutation and age are the only parameters with an independent impact on all outcome parameters (OS, EFS, RFS, CR). These data provide the basis for a prognostic model in NK-AML that can be used for risk stratification and selection of patients that will benefit from allogeneic stem cell transplantation.

A Combined Molecular and Clinical Prognostic Index For Relapse and Survival In Cytogenetically Normal AML (PINA)

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1303-1303
Author(s):  
Friederike Pastore ◽  
Annika Dufour ◽  
Tobias Benthaus ◽  
Klaus H. Metzeler ◽  
Kati Maharry ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Conflicts Of Interest ◽  
Clinical Care ◽  
Performance Status ◽  
Prognostic Index ◽  
Genetic Group ◽  
White Blood Count ◽  
Molecular Characteristics ◽  
Npm1 Mutation ◽  
Prognostic Indices

Abstract Background Cytogenetically normal acute myeloid leukemia (CN-AML) is the largest and most heterogeneous cytogenetic AML subgroup. For the practicing clinician it is difficult to know how to use the prognostic information of the growing number of clinical and molecular markers. Our purpose was to develop a widely applicable prognostic model by combining well-established pre-treatment patient and molecular characteristics. Patients and methods Two prognostic indices for CN-AML, one with regard to overall survival (PINAOS) and the other regarding relapse-free survival (PINARFS) were derived based on a cohort of 669 CN-AML patients treated within the AML Cooperative Group 99 (AMLCG99) study. Results Based on age (median: 60 years [range: 17-85 years]), performance status, white blood count, and presence or absence of NPM1 mutation, biallelic CEBPA mutation, and FLT3-ITD, patients were classified into three risk groups according to PINAOS and PINARFS: 29% of all and 32% of responding patients had low risk (5-year OS 72%; 5-year RFS 55%), 56% and 39% intermediate risk (5-year OS 28%; 5-year RFS 27%), and 15% and 29% high risk disease (5-year OS 3%; 5-year RFS 8%) (Figure 1). PINAOS and PINARFS further subdivided the European LeukemiaNet (ELN) favorable-genetic group as well as the ELN intermediate-I-genetic group. Both, PINAOS and PINARFS were confirmed in a large, independent, and comparable CN-AML cohort of 529 patients from the Cancer and Leukemia Group B (CALGB/Alliance) trials (Figure 2). Conclusions We have developed and validated the first prognostic indices specifically designed for CN-AML patients of all ages combining well-established molecular and clinical variables easily applicable in routine clinical care. The integration of both clinical and molecular markers could provide a basis for individualized patient care by risk-adapted therapy of CN-AML. Disclosures: No relevant conflicts of interest to declare.

Identification of Specific Genes Expression Signatures for CEBPa, NPM1 and FLT3 Genes Mutations in AML. A Study Based of the MILE Data Issued from 79 French Samples.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3481-3481
Author(s):  
Christophe Roumier ◽  
Olivier Nibourel ◽  
John Devos ◽  
Pacale Cornillet-Lefebvre ◽  
Sylvia Merk ◽  
...  
Keyword(s):  
Phase 1 ◽  
Gene Expression Signature ◽  
Normal Karyotype ◽  
Gene Signature ◽  
Molecular Data ◽  
P Value ◽  
Npm1 Mutation ◽  
Molecular Alterations ◽  
Flt3 Mutations

Abstract The MILE (Microarray innovations in Leukemia) project has recently reported on the efficiency and accuracy of micro-array use in the diagnosis and sub-classification of leukemia. The MILE algorithm classifier of AML recognized 6 subgroups according to major cytogenetic findings:t(15;17), t(8;21),inv(16), MLL rearrangement, complex karyotype and a last group including normal cytogenetics and other patients. In the latter group, beside age, leucocytosis and antecedent of previous hemopathy, molecular alterations, especially of CEBPa, NPM1 and FLT3 mutations have been shown to be frequent and to have a major prognosis value. In this work, we have attempted to enhance the MILE classification of AML by examining data for a gene signature specific of each of these mutations. The study was made using the data generated from HG-U133 plus 2.0 Affymetrix arrays from the 79 AML samples included from the French study centers of MILE phase 1 trial. Among these patients with AML, according to the gold standard classification used in the MILE project were 2 t(8;21) AML, 2 inv(16) AML and the others belonging to intermediate cytogenetic group class MILE 13 (mainly patients with normal karyotype). Into the cohort, 7 samples harbored CEBPa mutations, 33 samples have NPM1 mutation and 26 samples have FLT3-ITD. Data generated from GEP profiles were combined to these molecular data to obtain predictor of mutation status. Predictors were based on differential gene expression signature using SVM methods. Differentially expressed genes sets were obtained using volcano plot with p value of 0.05 and fold change of 2. The CEBPa predictor shows a sensitivity of 100% in the detection of CEBPa mutation with a specificity of 81%, only six sample were misclassified among them 3 were non informative and 3 were called as CEBPa mutated (2 t(8;21) samples and one AML with MLL alteration), the efficiency of the predictor was 83%. The NPM1 predictor shows a sensitivity of 87.9% in the detection of NPM1 mutation and a specificity of 75%, 7 samples were misclassified, among them 3 NPM1 mutated samples (2 were called as wt and one assigned as non informative) and at the other hand 4 NPM1 wt samples were called as mutated), efficiency of this predictor was 81%. To note using the set of genes used for this predictor, non supervised hierarchical tree clearly shows the dichotomy of NPM1 mutated samples into two sub-clusters according to the FLT3 status. Results were weaker for the FLT3-ITD predictor with a sensitivity of 57% and a specificity of 70% with 29 samples misclassified (9 miscalled and 20 without answer).These results shows that the use of the data generated by the MILE in the diagnosis of leukemia can be used in the detection of CEBPa, NPM1 and FLT3 mutations in AML that have a major value in the prognosis of patients. The use of this 3 new predictors, in second line after the use of MILE algorithm to discriminate AML belonging to the intermediate cytogenetic groups clearly improve the use of microarrays in the diagnosis of AML.

Microarray Analysis Detects Unique Expression Pattern for NPM1-Mutated AML with Normal Karyotype and Reveals Pathobiological Insights.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2382-2382
Author(s):  
Torsten Haferlach ◽  
Claudia Haferlach ◽  
Alexander Kohlmann ◽  
Lothar Wieczorek ◽  
Martin Dugas ◽  
...  
Keyword(s):  
Gene Expression ◽  
Melting Curve ◽  
Normal Karyotype ◽  
Support Vector ◽  
Melting Curve Analysis ◽  
Study Cohort ◽  
Data Set ◽  
Npm1 Mutation ◽  
Mutation Status ◽  
Mutational Status

Abstract Recent data indicate that mutations in exon 12 of the nucleophosmin (NPM1) gene characterize a distinct subgroup of adult and pediatric acute myeloid leukemia (AML). AML carrying NPM1 mutations account for about one-third of all adult AML, exhibit distinctive biological and clinical features and show a strong association to AML with normal karyotype (55% mutated). However, the role of NPM1 in leukemogenesis still remains elusive. Here we present data on a cohort of n=66 AML cases with normal karyotype analyzed by high-density whole genome expression microarrays (Affymetrix HG-U133 Plus 2.0). In parallel melting curve analysis was used to assess NPM1 mutational status: 41 cases were characterized as mutated (NPM1+) and 25 cases were unmutated (NPM1−). We first investigated the gene signature that discriminated NPM1+ from NPM1− cases. Genes that were significantly overexpressed comparing NPM1+ against NPM1– cases included a strong homeobox genes signature (HOXA1, HOXA5, HOXA7, HOXA9, HOXA10, HOXA11, HOXB2, HOXB4, HOXB5, HOXB6, HOXB7, MEIS1, and PBX3). A functional analysis (Gene Ontology) revealed a clear association of the group of overexpressed genes with the cell components nucleosome, chromatin, and the nuclear envelope-endoplasmatic reticulum network as well as involvement in the biological processes of nucleosome and chromatin assembly, establishment of protein transport and localization, and Notch signaling pathway. In contrast, the cellular processes completely differed when genes were investigated that were significantly underexpressed in NPM1+ cases compared to NPM1− cases. This group of genes encoded membrane-related proteins (gap junction, intercellular junction, signalosome complex) and proteins involved in cellular morphogenesis and cell communication. The differences in gene expression signatures between NPM1+ and NPM1− cases permit a robust classification approach by gene expression profiling. Support Vector Machine analysis resulted in &gt;92% prediction accuracy of NPM1 mutation status (10-fold cross-validation). The sensitivity was very high for the positive detection of NPM1+ cases (&gt;97%). Using a 100-fold re-sampling approach and splitting the complete data set into a training set (n=44) and testing set (n=22) the following genes were most frequently selected as top discriminatory genes: HOXA5, HOXB4, HOXB5, HOXB6, MEIS1, PBX3, FGFR1, ADAM17, PRICKLE1, and TMPO. Interestingly, the classification was less accurate when also FLT3 internal tandem duplication mutation status was taken into account. The study cohort (n=66) then was distributed as follows: 19 NPM1+/FLT3+, 22 NPM1+/FLT3−, 4 NPM1−/FLT3+, and 21 NPM1−/FLT3− negative cases. Only 14 of 22 (64%) NPM1+/FLT3– cases were correctly predicted, with miscalls falling both into the group of NPM1+/FLT3+ and NPM1−/FLT3− cases. In conclusion, NPM1 mutations are the most frequent mutations in adult AML to date and their central prognostic role is increasingly recognized. Given the fact that they are nearly mutually exclusive with major recurrent genetic abnormalities and that they can be characterized by a distinctive gene expression program these data especially for of NPM1+/FLT3− AML with better outcome may support to classify this as a separate biological subgroup of AML with normal karyotype.

IDH1/2, TET2 and DNMT3A Mutations Are Not Mutually Exclusive in Secondary Acute Myeloid Leukemias,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3556-3556
Author(s):  
Olivier Kosmider ◽  
Olivier LaRochelle ◽  
Marie-Magdelaine Coude ◽  
Veronique Mansat-De Mas ◽  
Eric Delabesse ◽  
...  
Keyword(s):  
Bone Marrow ◽  
De Novo ◽  
Conflicts Of Interest ◽  
Remission Rate ◽  
Normal Karyotype ◽  
Prognostic Impact ◽  
Mutational Status ◽  
Myeloid Leukemias ◽  
Melting Analysis ◽  
De Novo Aml

Abstract Abstract 3556 IDH1/2, TET2 and DNMT3A mutations have been reported in myeloid malignancies including de novo AML. In this study, we have analyzed the frequency and prognostic impact of these mutations in a large retrospective cohort of patients (pts) with secondary AML (SA) which encompass myelodysplasia-related changes (MRC) AML and therapy-related (TR) AML according to the WHO classification. Bone marrow samples were collected from 247 pts at diagnosis with SA and the mutational status of IDH1/2, TET2 and DNMT3A genes together with other genes frequently mutated in AML (NPM1, FLT-3, N and K-RAS, WT1) was determined by Sanger sequencing or high resolution melting analysis. The cohort of 247 pts consisted in 201 MRC AML and in 46 TR AML, 39.5% of which with a normal karyotype (NK). The frequency of IDH1/2, TET2 and DNMT3A mutations was 12.6, 19.8 and 4.5%, respectively. Two pts had both TET2 and IDH1/2 mutations, 2 pts had TET2 and DNMT3A mutations and 5 pts had both IDH1/2 and DNMT3A mutations showing that these mutations were not mutually exclusive in SA. IDH1/2 and TET2 mutations were significantly more frequent in MRC AML (14.1 and 22.3%) than in TR AML (6.4 and 8.7%) (P =0.04 and P =0.03) while the frequency of DNMT3A mutations was identical in the two subgroups. The SA pts harbouring at least one IDH1/2 or TET2 or DNMT3A mutation were significantly older (P <0.0001) and presented higher leukocyte count and lower MCV (P <0.05) than unmutated pts. Percentage of blasts in the bone marrow was similar in the two groups. Karyotype was normal in 48% of the IDH1/2 or TET2 or DNMT3A mutated pts and 18% of the unmutated patients, indicating that these mutations were strongly associated with NK (P < 0.001). A statistically significant link was found between TET2 or IDH1/2 or DNMT3A mutations and NPM1 mutations, but not with FLT-3, N/K-RAS or WT1 mutations. Complete remission rate and overall survival were evaluated in a group of 158 pts which had received intensive chemotherapy at diagnosis, and were identical in the IDH1/2 or TET2 or DNMT3A mutated and unmutated groups. These mutations did significantly influence survival neither in the subgroup of pts with normal karyotype, nor in the subgroup of MRC-AML, or TR-AML which were of very poor prognosis. These data show that IDH1/2, TET2 or DNMT3A mutations could modify the clinical presentation without impact on prognosis. Disclosures: No relevant conflicts of interest to declare.

A novel hierarchical prognostic model of AML solely based on molecular mutations

Blood ◽  
2012 ◽  
Vol 120 (15) ◽  
pp. 2963-2972 ◽  
Author(s):  
Vera Grossmann ◽  
Susanne Schnittger ◽  
Alexander Kohlmann ◽  
Christiane Eder ◽  
Andreas Roller ◽  
...  
Keyword(s):  
Myeloid Leukemia ◽  
Prognostic Model ◽  
Tp53 Mutation ◽  
Cox Regression ◽  
Normal Karyotype ◽  
Npm1 Mutation ◽  
Molecular Alterations ◽  
Kaplan Meier ◽  
Runx1 Mutation ◽  
Acute Myeloid

Abstract The karyotype is so far the most important prognostic parameter in acute myeloid leukemia (AML). Molecular mutations have been analyzed to subdivide AML with normal karyotype into prognostic subsets. The aim of this study was to develop a prognostic model for the entire AML cohort solely based on molecular markers. One thousand patients with cytogenetic data were investigated for the following molecular alterations: PML-RARA, RUNX1-RUNX1T1, CBFB-MYH11, FLT3-ITD, and MLL-PTD, as well as mutations in NPM1, CEPBA, RUNX1, ASXL1, and TP53. Clinical data were available in 841 patients. Based on Cox regression and Kaplan-Meier analyses, 5 distinct prognostic subgroups were identified: (1) very favorable: PML-RARA rearrangement (n = 29) or CEPBA double mutations (n = 42; overall survival [OS] at 3 years: 82.9%); (2) favorable: RUNX1-RUNX1T1 (n = 35), CBFB-MYH11 (n = 31), or NPM1 mutation without FLT3-ITD (n = 186; OS at 3 years: 62.6%); (3) intermediate: none of the mutations leading to assignment into groups 1, 2, 4, or 5 (n = 235; OS at 3 years: 44.2%); (4) unfavorable: MLL-PTD and/or RUNX1 mutation and/or ASXL1 mutation (n = 203; OS at 3 years: 21.9%); and (5) very unfavorable: TP53 mutation (n = 80; OS at 3 years: 0%; P < .001). This comprehensive molecular characterization provides a more powerful model for prognostication than cytogenetics.

The Impact of FLT3-ITD and NPM1 Mutational Status on the Outcome of ATRA Therapy in Patients with Non-APL AML: Results of the UK MRC AML12 Trial

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 554-554 ◽  
Author(s):  
Rosemary Gale ◽  
Robert Hills ◽  
Claire Green ◽  
Yashma Patel ◽  
Amanda Gilkes ◽  
...  
Keyword(s):  
Overall Survival ◽  
Normal Karyotype ◽  
Atra Treatment ◽  
Npm1 Mutation ◽  
Mutational Status ◽  
Hazard Ratios ◽  
The Uk ◽  
The Impact ◽  
To Receive

Abstract Background: The prognostic value of mutations in NPM1 and FLT3-ITD is well known. Previously, Schlenk et al. (ASH 2007, Abstract 297) have reported that survival was significantly improved in a group of older patients who were NPM1 mutant/FLT3-ITD Wild Type when treated with ATRA; there was no significant improvement in overall survival in patients who were either NPM1 WT or FLT3-ITD mutant. We report here on a subset of the UK MRC AML12 trial who have been characterised for FLT3-ITD and NPM1 and who were randomised between ATRA in induction and no ATRA. Patients and Methods: A total of 393 patients were identified and characterised. The median age was 46 years (range 16–68); NPM1 and FLT3-ITD status were determined using methods previously reported (Gale et al. Blood 2008). Median follow-up for survival is 7.1 years. The overall results of the ATRA randomisation have previously been reported (Burnett et al. ASH 2002 Abstract 529) and show no benefit for ATRA treatment. All patients were treated with Daunorubicin, Ara-C and Thioguanine (DAT) with a randomisation between two doses of Ara-C, and were randomised to receive, or not, ATRA 45mg/m2/d during courses 1 and 2 of chemotherapy. ATRA was given for a median of 56 days. Results: A total of 143 (36%) patients had an NPM1 mutation and 93 (24%) had a FLT3-ITD mutation. No significant interactions were seen between either NPM1 status, or FLT3-ITD status and ATRA treatment with respect to complete remission, overall survival or relapse free survival (see Table). Estimates of the hazard ratios (HR) for the interaction between FLT3-ITD and ATRA, and NPM1 and ATRA for overall survival were 0.75 (95% CI 0.42–1.32 p=0.3) and 0.66 (95% CI 0.38–1.12 p=0.13), where an HR&lt;1 indicates greater benefit for ATRA in the mutant group. Looking at patients stratified by both FLT3-ITD and NPM1 status (84 NPM1+ITD−, 34 NPM1−ITD+, 59 NPM1+ITD+, 216 NPM1−ITD−) showed no significant interaction (p=0.4 for heterogeneity of ATRA effect between the four groups, p=0.5 for difference in treatment effect between FLT3 WT/NPM1 mutant and others). The results were not significantly different if restricted to patients with a normal karyotype only. Conclusions: In this randomised comparison of ATRA therapy in younger patients with AML there were no significant interactions. Any impact of NPM1 or FLT3-ITD status on treatment with ATRA is likely to be relatively small or non-existent. CR OS at 5years RFS at 5 years ATRA No ATRA OR, 95% CI ATRA No ATRA HR, 95% CI ATRA No ATRA HR, 95% CI NPM1 WT 80% 86% 1.59 (0.82–3.09) 36% 35% 1.07 (0.79–1.45) 33% 29% 1.05 (0.76–1.46) NPM1 Mutant 91% 89% 0.81 (0.27–2.43) 57% 44% 0.70 (0.45–1.11) 49% 39% 0.83 (0.63–1.31) Interaction with ATRA p=0.3 0.1 0.4 FLT3 WT 83% 87% 1.46 (0.77–2.74) 45% 42% 0.97 (0.73–1.30) 42% 35% 0.87 (0.64–1.18) FLT3 ITD 89% 88% 0.89 (0.25–3.17) 40% 25% 0.73 (0.44–1.21) 32% 26% 0.89 (0.53–1.50) Interaction with ATRA p=0.5 0.4 0.9

Expression of KDR Plus Detection of Flt3-ITD and NPM1 Mutation as a Prognostic Marker in AML Patients with Normal Karyotype.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4715-4715
Author(s):  
Aibin Liang ◽  
Bing Xiu ◽  
Binbin Huang ◽  
Ying Han ◽  
Lanjun Bo
Keyword(s):  
Fungal Infection ◽  
Outcome Prediction ◽  
Conflicts Of Interest ◽  
Normal Karyotype ◽  
High Dose ◽  
Npm1 Mutation ◽  
Positive Expression ◽  
High Dose Cytarabine ◽  
Cytogenetic Analyses

Abstract Abstract 4715 From July 2006 to February 2009, total of 146 patients with non-APL AML were admitted in our department. Cytogenetic analyses, detection of Flt3-ITD, NPM1 gene mutation and VEGF and its receptors (Flt-1, KDR) were performed on all patients. Forty-nine patients less than 60 years of old and with normal karyotype were selected for prognostic analyses. Flt3-ITD was positive detected in 15 cases (30.61%), NPM1 mutation in 18 cases (36.73%), VEGF in 46 cases (93.88%), Flt-1 in 41 patients (84.04%) and KDR in 38 cases (77.55%). After one or two courses of induction therapy with IDA regimen (Idarubicin + cytarabine: 3+7) in all 49 patients, total CR rate was 67.43% (33/49). One patient died because of severe invasive fungal infection. Among the remaining 15 non-CR patients, 10 were Flt3-ITD positive and NPM1 negative, and all with higher expression of VEGF and KDR. All the CR patients were treated with consolidation regimen with high dose cytarabine (3g/m2, q 12h iv for 3 consecutive days) for 6 courses. After follow-up at least for 6 months, only 12 patients are alive up to now. In these 12 patients, Flt3-ITD were all negative expressed, 6 patients were NPM1 positive, 2 patients VEGF negative, 3 patients both KDR and Flt-1 negative. There are no patients alive with positive expressed of Flt3-ITD, KDR and negative expressed of NPM1. Therefore, we supposed that negative expression of Flt3-ITD and KDR plus positive expression of NPM1 could be a favorable parameter for outcome prediction in AML patients with normal karyotype. Disclosures: No relevant conflicts of interest to declare.

Costimulatory Expression Profiling of AML Blasts Identifies Surface Markers with High Correlation to Isolated NPM1 Mutation

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 2511-2511
Author(s):  
Felix S Lichtenegger ◽  
Michael Krempasky ◽  
Karsten Spiekermann ◽  
Jan Braess ◽  
Wolfgang Hiddemann ◽  
...  
Keyword(s):  
Stem Cell ◽  
Molecular Markers ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Expression Pattern ◽  
Allogeneic Stem Cell Transplantation ◽  
Risk Group ◽  
Npm1 Mutation ◽  
Mutational Status ◽  
Blast Cells

Abstract Abstract 2511 Acute myeloid leukemia (AML) is a heterogeneous disease. In recent years, numerous cytogenetic and molecular markers providing prognostic information have been established. The success of allogeneic stem cell transplantation and some autologous immunotherapeutic strategies has proven that immunological effects play an important role for treatment of AML, especially for the eradication of minimal residual disease. However, the effect is dampened by immunosuppressive factors provided by AML blasts. One of the possible immunomodulatory mechanisms is the expression pattern of costimulatory and coinhibitory molecules on AML cells, which influences their interaction with specific T cells. In order to elucidate the potential significance of this mechanism in AML, we analyzed the surface expression of a broad panel of costimulatory and coinhibitory molecules (CD80, CD86, CD273, CD274, CD275, CD276, B7-H4, HVEM) on blast cells of 102 AML patients at initial diagnosis by flow cytometry. The mean fluorescence intensity of these markers on the CD33 positive blast cell population was correlated with morphologic, cytogenetic and molecular characteristics of the disease. Statistical significance of differences in expression levels was tested with the Mann-Whitney-U test for independent samples. First, we could show that the AMLs morphologically classified as M4 or M5 according to FAB (n = 35) expressed higher levels of costimulatory markers, especially CD86 (p < 0.001) and CD276 (p = 0.016), compared to the other morphologic subgroups. For CD86, this is in concordance with the literature (e.g. Maeda et al., Br J Haematol 1998). Correlation with CD276 has not been published before, but is well in line with the monocytic lineage of these AML cases, inclining them to good antigen presentation capacities. In contrast to morphologic characteristics, correlations of the AML costimulatory profile with cytogenetic and molecular markers of the disease have never been studied before. AML patients were classified according to ELN prognostic groups, which are based on cytogenetic and molecular markers (Döhner et al., Blood 2010). Interestingly, we found that the AML blast cells of patients allocated to the favorable risk group (n = 23) showed higher expression of CD86 (p = 0.049), CD274 (p = 0.010), CD276 (p = 0.003), and B7-H4 (p = 0.001), compared to patients of the non-favorable risk groups. In particular, the cases of leukemia with normal karyotype and an isolated NPM1 mutation (without accompanying FLT3-ITD mutation), which constituted about half of the favorable risk group (n = 11), were responsible for this pattern. In these cases, the balance of costimulatory and coinhibitory molecules was clearly shifted toward expression of positive costimulatory molecules, reflected by a significantly higher ratio of the positive costimulatory molecule CD86 to various coinhibitory molecules (e.g. p = 0.003 for the ratio CD86/CD274). To further dissect the influence of NPM1 mutation on the costimulatory expression profile, all patients with an NPM1 mutation irrespective of FLT3 mutational status and karyotype were analyzed. In this cohort of 33 patients, a similar trend to higher expression of these molecules was observed, but correlations were less pronounced (p = 0.012 for CD86) or not significant (CD274, CD276, B7-H4). The karyotype and the FLT3 mutational status by itself, however, showed no significant correlation with the costimulatory profile. We hypothesize that the expression pattern of costimulatory molecules contributes to prognosis of blast clearance and especially relapse. If this is the case, we are interested to analyze whether the marker profile constitutes a surrogate for molecular markers or an independent prognostic marker. Potentially, this could be highly relevant for application in immunotherapy, e.g. allogeneic stem cell transplantation. A preliminary analysis of the clinical data will be presented. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Frequent Mutations and Their Influence on Prognosis in Patients with Acute Myeloid Leukemia

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 4949-4949
Author(s):  
Ekaterina Petrova ◽  
Irina Martynkevich ◽  
Luybov Polushkina ◽  
Luydmila Martynenko ◽  
Marina Ivanova ◽  
...  
Keyword(s):  
De Novo ◽  
Conflicts Of Interest ◽  
Prognostic Significance ◽  
Melting Curve ◽  
Normal Karyotype ◽  
Genetic Alterations ◽  
Melting Curve Analysis ◽  
Prognostic Impact ◽  
Age Related ◽  
Mutational Status

Abstract Background. Several genetic alterations such as translocations, gene mutations and deletions play an important role in myeloid leukemogenesis. The cytogenetic information is a very significant tool to classify pts at their initial diagnosis into prognostic categories. For pts with cytogenetically normal AML, prognosis can be specified by mutational status of the genes NPM1, FLT3, CKIT, NRAS and DNMT3A. The aim of the research was to investigate the frequency and prognostic impact of FLT3, NPM1, CKIT, NRAS and DNMT3A mutations in AML pts and to analyze their interaction with other prognostic markers. Methods. This study was performed in 200 adult pts (190 pts with de novo and 10 pts with secondary AML), previously untreated, median age 55 years (18-86). According to the results of cytogenetic analyses pts were separated in four groups: with favourable (9,0%), unfavourable (14,0%) prognosis, with normal karyotype (NK) (48,5%) and other aberrations (28,5%). Mutations in FLT3, CKIT and NPM1 were analysed by PCR and in NRAS by sequencing. Mutation analysis of DNMT3A R882 was performed by high-resolution melting curve analysis. Cytogenetic studies were analysed on bone marrow samples using standard GTG-method. Results. Mutations in FLT3, CKIT, NRAS and NPM1 genes were detected in 105/200 (52,5%) pts. A total of 128 mutations were revealed in this group: 24,0% - FLT3-ITD, 6,5% - FLT3-TKD, 20,5% - in NPM1, 10,0% - in NRAS and 3,0% - in CKIT. 82 pts had single mutations and in 23 pts mutations occurred simultaneously: 17 with FLT3-ITD and in NPM1, 2 with FLT3-ITD and FLT3-TKD, 1 with FLT3-TKD and in NPM1, 3 with NPM1 and NRAS mutations. We found that mutations with the significantly higher incidence (p=0,001) were observed in the group of pts with NK (80/97), whereas there were only 8/28 pts with mutations in the group with complex karyotype. When analyzing the age-related features, it was shown that the majority of mutations were detected in the group of pts at the age from 60 to 69 years. Mutations FLT3-ITD and FLT3-TKD were associated with higher WBC count comparing with pts without mutations (p=0,001 and p=0,014, respectively). The median follow-up for overall (OS) and relapse-free (RFS) survival for pts with FLT3-ITD against ptswith FLT3-ITD- was: 5,4 vs 12,8 months and 4,9 vs 10,0 months (p=0,001 and p=0,001), respectively. Mutation FLT3-TKD was also found to be prognostically unfavorable, but only comparing with pts with FLT3-ITD- genotype. As the result of OS and RFS analyses in pts with and without NPM1 mutations we revealed the significant favorable influence of NPM1 mut on the prognosis (p=0,002 and p=0,020, respectively). However pts with genotype FLT3-ITD+/NPM1+ were found to get to the group with an intermediate risk. We detected the significant adverse influence of CKIT mut on RFS (p=0,041). Mutations in NRAS didn't impact on prognosis; we only showed the tendency towards worsening of OS and RFS in group of pts with favorable cytogenetics (p=0,214 and p=0,160, respectively). Mutations DNMT3A R882 were investigated in group of 143 AML pts and were detected in 23 (16,1%) pts. Pts with DNMT3A R882 had higher WBC (p=0,001) and platelets (p=0,020) count at diagnosis and more frequently belonged to FAB groups M5 (p=0,003), as compared with DNMT3A wt. Of 23 pts who had AML with DNMT3A mutations, 17 had tumors with NK profiles (24,3% of a total of 70 cytogenetically normal samples) (p=0,009). Pts with isolated DNMT3A mutations were seen in 4 cases, whereas in the rest of pts they were detected simultaneously with mutations in genes FLT3, NPM1, NRAS and CKIT. DNTM3A mutations were significantly more prevalent in NPM1 mut (p=0,005) and FLT3-ITD (p=0,005) positive cases than wild type. DNMT3A mutations were associated with negative influence on pts OS and risk of relapse, compared with DNMT3A wt (р = 0,031 and р = 0,045, respectively). Summary. Mutations in FLT3 and NPM1 had a significantly higher incidence in the group of pts with a normal karyotype. FLT3 mutations showed the adverse prognostic value. Insertions in NPM1 were shownto be the favorable factor, correlating with prolonged RFS in all pts excepting pts with FLT3-ITD+/ NPM1+ genotype. CKIT mut was associated with higher relapse incidence in AML pts, while NRAS mut showed lack of prognostic significance. AML with DNMT3A mut represent the group, homogeneous on a number of clinical and laboratory parameters, associated with adverse prognosis and a high risk of the relapse. Disclosures No relevant conflicts of interest to declare.

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