scholarly journals H3K27me3 Level of the HIST1 Cluster Defines an Epigenetic Marker of Acute Myeloid Leukemia with Prognostic Value

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 2326-2326
Author(s):  
Guillaume Tiberi ◽  
Aleksandra Pekowska ◽  
Claire Oudin ◽  
Adam Ivey ◽  
Thomas Prebet ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Prognostic Value ◽  
Molecular Characteristics ◽  
Significant Heterogeneity ◽  
Npm1 Mutation ◽  
Clinical Prognosis ◽  
H3k27me3 Level ◽  
The Impact ◽  
Acute Myeloid

Abstract Acute myeloid leukemia (AML) is the most common acute leukemia diagnosed in adults, characterized by significant heterogeneity in terms of biology and clinical outcome. Improvements in sequencing technologies have led to the discovery of frequent somatic mutations in epigenetic modifiers, placing epigenetic deregulation in the center of AML. Yet, the global view and the impact of this deregulation on disease characteristics are under investigation. To gain a more comprehensive understanding of epigenetic deregulation in AML, particularly the heterogeneous subset with normal karyotype (CN-AML), associated with intermediate clinical prognosis, we performed H3K27me3 profiling on CN-AML patient samples. Primary bone marrow or peripheral blood samples containing more than 80% of blasts were selected from the Institut Paoli-Calmettes Biological Resources Center inventory for the purpose of genetic and epigenetic studies. We initially analyzed 35 CN-AML samples by ChIP coupled with hybridization on oligonucleotide promoter arrays (Chip-chip) for genomic H3K27me3 distribution. Clustering analysis revealed 586 highly H3K27me3-variable genomic regions across patients corresponding to 461 genes mostly involved in chromatin organization. The heterogeneity in the H3K27me3 profile was characterized by a remarkable H3K27me3 enrichment at the chromosome 6 p22.2-22 region that encompasses 70 kbp within the major HIST1 cluster. This striking H3K27me3 enrichment was covering 11 histone genes and was partially overlapping with the focal deletion at 6p22 found in acute lymphoblastic leukemia. The HIST1 H3K27me3 enrichment profile clearly distinguished 2 groups of CN-AML patients based on their HIST1 H3K27me3 level. In order to independently extend this observation, we analyzed the H3K27me3 status by using ChIP followed by qPCR (ChIP-qPCR) at the same HIST1 genomic locations, in an independent cohort of 51 CN-AML patients. This revealed the presence of this abnormal epigenetic profile in about 50% of the patients. CN-AML samples were split in two groups, according to the median H3K27me3 enrichment levels at the HIST1 cluster genes. These two groups were analyzed for clinical and molecular characteristics. Patients with high HIST1 H3K27me3 level had a markedly higher incidence of NPM1 mutation (89% vs. 40%; p= 1.75x10-5) and a lower incidence of WT1 mutation (0% vs. 20%; p=0.028). No significant association was observed with FLT3 (ITD and TKD), IDH1/2, DNMT3A nor ASXL1 mutations. Patients with high HIST1 H3K27me3 level had a significant longer leukemia free survival at 5 years (allo-grafted patients censored, LFS-allo; 13.33 vs. 8.92 months p=0.0053). Moreover, multivariate analysis showed that HIST1 H3K27me3 status provided a more powerful prognostic indicator than the NPM1mut/FLT3-ITDneg and NPM1/FLT3-ITD genotypes. In conclusion,using epigenetic profiling, our analysis has enabled the discovery of a new epigenetic alteration that affects CN-AML and impacts prognosis. We demonstrate that the HIST1 cluster is targeted by epigenetic events that lead to high H3K27me3 level and predicts a good prognosis. This may help refine risk stratification in AML, identifying a further group of patients unlikely to benefit from allogeneic transplantation in first remission. Overall, our data provide a proof of concept that epigenetic profiling could be used to discover new biomarkers with prognostic value. Disclosures No relevant conflicts of interest to declare.

scholarly journals The LincRNA HOTAIRM1, Located in the HOXA genomic Region, impacts Prognosis in Acute Myeloid Leukemia and Is Associated with a Distinctive microRNA Signature

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 1003-1003
Author(s):  
Marina Díaz-Beyá ◽  
Alfons Navarro ◽  
Salut Brunet ◽  
Josep F Nomdedeu ◽  
Anna Cordeiro ◽  
...  
Keyword(s):  
Gene Expression ◽  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Prognostic Value ◽  
Hox Genes ◽  
Genomic Region ◽  
Expression Level ◽  
Molecular Characteristics ◽  
Npm1 Mutation ◽  
Acute Myeloid

Abstract Background: Non-coding RNAs (ncRNAs) have recently emerged as key regulators of diverse cellular processes, including leukemia. ncRNAs are classified according to their size as short (eg, microRNAs) or long ncRNAs. lincRNAs are long ncRNAs located in intergenic regions and have multiple regulatory functions, including gene expression regulation. Interestingly, active crosstalk between microRNAs and lincRNAs has been observed. lincRNAs are known to be deregulated in some cancers but their importance in acute myeloid leukemia (AML) is so far unknown. HOX genes play an important role in hematopoiesis and are deregulated in AML. lincRNAs are especially abundant in the clusters of HOX genes. HOTAIRM1, a myeloid lineage-specific lincRNA, is located at the 3’end of the HOXA cluster and seems to play a regulatory role in myelopoiesis. However, to date the potential prognostic role of HOTAIRM1 expression in AML has not been examined. Aims: To investigate first whether the expression of the lincRNA HOTAIRM1 is associated with the clinical, cytogenetic and molecular characteristics and microRNA expression in AML patients. Secondly, since intermediate risk (IR) AML patients have a highly diverse prognosis, we analyzed the potential prognostic value of HOTAIRM1 expression in IR-AML patients. Methods: To explore the expression level of HOTAIRM1 among different AML subtypes, we analyzed samples from 244 AML patients including CBF-rearranged AML (n=5), APL (n=4), MLL-rearranged AML (n=3), EVI1-rearranged AML (n=3), t(6;9) AML (n=9), AML with monosomal karyotype (n=3), and a large cohort of IR-AML (described below). For the analysis of prognostic value of HOTAIRM1, we analyzed specifically the outcome of 217 IR-AML patients (median age, 52; 51% males) sequentially included in CETLAM trials during the period 1995-2009. Molecular genotyping of this group identified NPM1 mutation (NPM1mut), FLT3-ITD, and biallelic CEBPA mutation (CEBPA mut) in 99 (45%), 79 (36%) and 17 (11%), respectively. The expression of HOTAIRM1 was analyzed using TaqMan® Gene Expression Assays (Applied Biosystems). microRNA and mRNA expression data were obtained in previous studies (Díaz-Beyá, Leukemia 2013). Statistical analyses were performed with BRB Array Tools, SPSS v20 and R v3.0. MaxStat package from R software was used to determine the optimal cutoff point of HOTAIRM1 expression. Results: Among all 244 patients, HOTAIRM1 expression was significantly different among the 7 included genetic subgroups (ANOVA p=0.0024), with the lowest levels observed in APL-AML patients and the highest in the t(6;9)AML patients. Within the IR-AML group, HOTAIRM1 overexpression was observed in NPM1mut patients (p<0.001). The prognostic study showed that high HOTAIRM1 expression was associated with shorter 5-year overall survival (OS) (27+11% vs.47+8%; p=0.009) shorter 5-year disease-free survival (LFS) (22+12% vs. 53+9%; p<0.001), and a higher cumulative incidence of relapse (CIR) at 5 years (55+15% vs. 34+8%; p=0.004). The effect on outcome was maintained within the subgroup with favorable molecular features (i.e., NPM1mut and CEBPAmut without FLT3-ITD) (OS: 75+11% vs. 39+29%; p=0.026). In the multivariate analysis including age, sex, WBC, NPM1mut, FLT3-ITD and number of treatment cycles for CR achievement as covariates, HOTAIRM1 expression emerged as an independent prognostic marker in OS (HR=2.44; 95% CI: 1.51-3.93; p<0.0001), LFS (HR=2.07; 95% CI: 1.31-3.24; p=0.002) and CIR (HR=2.05; 95% CI: 1.18-3.55; p=0.01). Supervised analysis by means of t-test based on multiple permutations revealed a distinctive 33-microRNA signature which correlated with HOTAIRM1 expression including miR-196b (p<0.001) located in the HOXA genomic region. Moreover, we correlated the expression of HOX genes and HOTAIRM1 and observed a positive correlation with HOXA4 gene expression (R2= 0.6; p=0.001). Conclusion: The expression level of the lincRNA HOTAIRM1 varied among different molecularly-defined AML. Interestingly, HOTAIRM1 expression level showed independent prognostic value within the IR-AML group. Moreover, HOTAIRM1 expression strongly correlates with its neighboring HOXA4 gene and harbors a distinctive microRNA signature. Our findings can pave the way for further studies of HOX-related lincRNAs and microRNAs regulatory networks and their influence on clinical outcome. Acknowledgments: ISCIII RH13/00205, SEHH, FIS13/00999 Disclosures No relevant conflicts of interest to declare.

Characterization of Extramedullary Acute Myeloid Leukemia – Results of the AML96 Trial

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2154-2154
Author(s):  
Friedrich Stölzel ◽  
Christoph Röllig ◽  
Michael Kramer ◽  
Brigitte Mohr ◽  
Uta Oelschlägel ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Acute Myeloid Leukemia ◽  
Induction Chemotherapy ◽  
Myeloid Leukemia ◽  
The Body ◽  
Hematopoietic Stem ◽  
Npm1 Mutation ◽  
Mutation Status ◽  
The Impact ◽  
Acute Myeloid

Abstract Abstract 2154 Background: Myeloid Sarcoma (MS) is defined as an extramedullary mass composed of myeloid blasts occurring at an anatomical site other than the bone marrow. Furthermore, the term extramedullary manifestation (EM) is applied if it accompanies overt acute myeloid leukemia (AML) and represents non-effacing tissue infiltration. EM is reported to correspond often to the skin but can affect almost every site of the body. The prognosis of MS or EM has been discussed controversially in the past. EM at diagnosis of AML is generally thought to be a rare event. However, data defining the prevalence of EM at diagnosis of AML and its prognostic value are missing. The aim of this analysis was to provide data for estimating the prevalence of EM at diagnosis of AML and to determine its relevance by including clinical and laboratory data from patients being treated in the prospective AML96 trial of the Study Alliance Leukemia (SAL) study group. Patients and Methods: A total of 326 patients with AML (age 17 – 83 years) and EM were treated within the AML96 trial with a median follow up of 8.8 years (95% CI, 8.4 to 9.3 years). All patients received double induction chemotherapy. Consolidation therapy contained high-dose cytosine arabinoside and for patients ≤ 60 years of age the option of autologous or allogeneic hematopoietic stem cell transplantation (HSCT). Logistic regression analyses were used to identify prognostic variables for CR rates. The method of Kaplan-Meier was used to estimate OS and EFS. Confidence interval (CI) estimation for the survival curves was based on the cumulative hazard function using the Greenwood's formula for the SE estimation. Survival distributions were compared using the log rank test. Results: 17% of the AML patients entered into the AML96 trial were diagnosed with EM. In 313 of the 326 patients (96%) EM was evident at diagnosis. The majority of patients with EM were diagnosed with de novo AML (84%, n=273), whereas gingival infiltration (51%, n=166) displayed the main EM of AML with CNS involvement being less common (4%, n=14). The majority of patients had a cytogenetic intermediate risk profile (71%, n=221) with a total of 172 patients (56%) harboring a normal karyotype. Patients with EM had a statistically significant lower median CD34-positivity of bone marrow blasts, higher percentage of FAB subtypes M4 and M5, higher WBC counts and LDH at diagnosis and higher percentage of NPM1 mutations compared to those patients without EM (all p<.001). When comparing achievement of CR between patients with EM to patients without EM, no statistical difference between these two groups was observed. Analysis according to the NPM1/FLT3-ITD mutation status revealed highest 5-year-OS (37%, 95% CI: .24 - .508) and 5-year-EFS (36%, 95% CI: .224 - .448) in the NPM1-mut/FLT3-wt group and lowest 5-year-OS (12%, 95% CI: 0 - .261) and 5-year-EFS (4%, 95% CI: 0 - .124) in the NPM1-wt/FLT3-ITD group, p=.007 and p=.001, respectively. Of the 49 relapsed patients with EM who had a NPM1-mutation at diagnosis 48 deceased despite of intensified relapse therapies. Conclusions: This analysis represents the largest study so far investigating the impact of EM AML. Patients with EM AML have distinct differences from AML patients without EM regarding their clinical and molecular characteristics at diagnosis. However these differences do not translate into differences in response to induction chemotherapy. Compared to patients without EM, survival analysis revealed differences according to the NPM1/FLT3-ITD mutation status which is also described for patients without EM AML. However, the prognosis for patients with EM who harbor a mutated NPM1 the prognosis at relapse seems to be dismal. Disclosures: No relevant conflicts of interest to declare.

Prognostic Value of Immunophenotyping in Elderly Patients with Acute Myeloid Leukemia: A Single Institution Experience.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4353-4353
Author(s):  
Claudiu Plesa ◽  
Youcef Chelghoum ◽  
Adriana Plesa ◽  
Mohamed Elhamri ◽  
Isabelle Tigaud ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Prognostic Factor ◽  
Elderly Patients ◽  
Myeloid Leukemia ◽  
Prognostic Value ◽  
Antigen Expression ◽  
Risk Groups ◽  
Significant Prognostic Factor ◽  
The Impact ◽  
Acute Myeloid

Abstract The poor prognosis of elderly patients with acute myeloid leukemia (AML) raises the question about the benefit of intensive chemotherapy in these patients. The impact of initial characteristics on prognosis has previously been addressed in elderly paitents, but very few data are available regarding the prognostic value of immunophenotypic characteristics in this setting. We investigated the expression of the membrane antigens CD13, CD15, CD33, and CD34 by flow cytometry in elderly patients with newly diagnosed AML, and analyzed whether these parameters possessed clinical or prognostic relevance, in order to help physicians in their choice of therapy. Immunophenotyping was performed in 273 patients aged 60 years or more (median 69 years). CD13 was expressed in 73%, CD15 in 43%, CD33 in 64%, and CD34 in 66% of cases. Complete remission was obtained in 157 cases (58%). The median overall survival was 8.1 months with a 3-year survival rate of 14%. Three risk groups were defined based on CD34 and CD33 antigen expression: Poor risk in patients with CD34+ CD33+ or CD34− CD33− disease, intermediate risk in patients with CD34+ CD33− disease, and favorable risk in patients CD34− CD33+ disease. Immunophenotype was, after cytogenetics, the most significant prognostic factor in terms of survival in a multivariate analysis (p = 0.03 and p < 0.0001 respectively). When combining immunophenotypic and cytogenetic parameters, patients were classified into four prognostic groups: Group A (3-year survival: 33%) including favorable and normal karyotypes with a favorable immunophenotype; Group B (3-year survival: 28%) including normal karyotypes with an intermediate immunophenotype; Group C (3-year survival: 8%) including intermediate or normal karyotypes with an unfavorable immunophenotype; and Group D (3-year survival: 2%) including all unfavorable cytogenetics. Immunophenotypic characteristics appeared to be a major prognostic factor in this patient population. Using two simple parameters assessed at time of diagnosis, we devised a prognostic system of immediate clinical utility for prognostic stratification and risk-adapted therapeutic choices.

The impact of FLT3 internal tandem duplication mutant level, number, size, and interaction with NPM1 mutations in a large cohort of young adult patients with acute myeloid leukemia

Blood ◽  
2008 ◽  
Vol 111 (5) ◽  
pp. 2776-2784 ◽  
Author(s):  
Rosemary E. Gale ◽  
Claire Green ◽  
Christopher Allen ◽  
Adam J. Mead ◽  
Alan K. Burnett ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Young Adult ◽  
Myeloid Leukemia ◽  
Tandem Duplication ◽  
Large Cohort ◽  
Internal Tandem Duplication ◽  
Npm1 Mutation ◽  
Beneficial Impact ◽  
The Impact ◽  
Acute Myeloid

An internal tandem duplication in the fms-like tyrosine kinase 3 gene (FLT3/ITD) is associated with poor prognosis in acute myeloid leukemia (AML), but the impact of mutant level, size, and interaction with nucleophosmin 1 (NPM1) mutations remains controversial. We evaluated these characteristics in a large cohort of young adult AML patients. There was a highly significant trend for worsening in relapse risk (RR) and overall survival (OS) with increasing FLT3/ITD mutant level (P < .001 for both), and even in the low level mutant group (1%-24% of total FLT3 alleles), RR was significantly worse than in the FLT3 wild-type (WT) group (P < .001). In multivariate analysis, mutant level was the most powerful prognostic factor for RR. Mutant size and number had no significant impact on outcome. The beneficial impact of an NPM1 mutation on RR and OS was seen in FLT3/ITD+ as well as FLT3/WT patients; both markers were highly significant independent predictors of outcome (P < .001). Stratification using both markers identified 3 prognostic groups: good (FLT3/ITD−NPM1+), intermediate (FLT3/ITD−NPM1− or FLT3/ITD+NPM1+), and poor (FLT3/ITD+NPM1−). Patients with high FLT3/ITD mutant level (greater than 50%) or FLT3/ITD+ in the absence of an NPM1 mutation may be good candidates for more experimental therapeutic approaches.

Prospective Evaluation of Immunophenotype In Correlation to Genotype In Previously Untreated Adult Patients with Acute Myeloid Leukemia In the Context of the WHO Classification

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1693-1693
Author(s):  
Michela Tassara ◽  
Konstanze Döhner ◽  
Christine Schöpflin ◽  
Anke Schmid ◽  
Heinz A. Horst ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Research Funding ◽  
Specific Treatment ◽  
Phase Iii ◽  
Molecular Characteristics ◽  
Acute Leukemias ◽  
Npm1 Mutation ◽  
Acute Myeloid ◽  
Who 2008

Abstract Abstract 1693 Background: The WHO 2008 classification of hematological malignancies defines distinct acute myeloid leukemia (AML) entities based on cytogenetic and molecular characteristics. However, immunophenotypotyping and morphology are still essential for rapid and correct diagnosis and for guidance of initial treatment, which in the future will be more and more genotype-specific. Although associations between phenotype and genotype have been described, correlations were evaluated mostly in retrospective studies not considering more recently identified gene mutations. Aims: To identify correlations between genotype and phenotype in adult AML. Methods: Immunophenotyping, cytogenetics and molecular analyses were performed centrally within the diagnostic screening procedure of a prospective multicenter phase III treatment trial for patients with AML aged 18 to 59 years. A total of 270 samples were analysed. The antibody panel for immunophenotyping was based on the recommendations of the German network of competence chronic and acute leukemias and the WHO-2008 classification. Results: AML with NPM1 mutation (NPM1mut) were characterized by higher expression of membrane-bound (m) CD33 (98% vs. 83%, p<0.0001), mCD11b (90% vs. 36%, p<0.0001) and mCD184 (90% vs. 37%, p<0.0001), whereas mCD34 (14% vs. 82%, p<0.0001), cytoplasmatic (c) CD34 (25% vs. 85%, p<0.0001), mHLA-DR (70% vs. 85%, p=0.001), mCD117 (66% vs. 86%, p<0.0001), and mCD133 (8% vs. 40%, p<0.0001) were significantly lower expressed compared with NPMwt AML. In AML with NPM1mut, the presence of an activating FLT3 mutation was associated with higher mCD34 (22% vs. 3% p=0.014), cCD34 (33% vs. 13% p=0.03) and lower mCD61 (11% vs. 31% p=0.03) and mCD42b (0% vs. 14% p=0.04) expression. AML carrying a CEBPA double-allelic mutation (double-CEBPAmut) was characterized by higher expression of mCD34 (100% vs. 56%, p<0.0001), cCD34 (100% vs. 62%, p=0.001), mCD117 (100% vs. 79%, p=0.003) and of the lymphoid-associated antigen mCD7 (93% vs. 27% p<0.0001) compared to single-CEBPAmut and CEBPAwt. This phenotype was strongly related to double-CEBPAmut (chi square test p<0.0001) but not to AML with single-CEBPAmut. AML with t(8;21) presented with a characteristic phenotype consisting of high cCD34 (100% vs. 62%, p=0.0009), mCD34 (100% vs.56% p=0.0001), mHLA-DR (100% vs.63% p=0.02), mCD117 (100 vs. 78%, p=0.03), and lymphoid-associated antigens mCD19 (76% vs. 2% p<0.00001) and mCD56 (64% vs.11% p< 0.00001) expression. AML with inv(16) or t(16;16) had higher expression of cMPO (100% vs. 88%, p=0.02), mCD34 (90% vs 60%, p=0.0002), cCD34 (86% vs. 65%, p=0.01) and mHLA-DR (96% vs. 81%, p=0.01), whereas lymphoid antigens and platelet-associated antigens were significantly lower expressed (mCD56, 0% vs. 17%, p=0.01; mCD7 3% vs. 35%, p=0.0002; mCD61, 0% vs. 20%, p=0.003; mCD41, 9% vs. 38%, p=0.008; mCD42b, 0% vs. 9%, p=0.04). The associations of specific phenotypes to genotypes were as follows: i) cMPOpos/mCD19pos/cCD34pos/mCD34pos/mCD117pos/mHLA-DRpos to t(8;21) (p<0.0001; sensitivity, 76%; specificity, 99%); ii) cMPOpos/mCD7pos/mCD34pos/cCD34pos/mCD117pos to double-CEBPAmut (p<0.0001; sensitivity, 94%; specificity, 90%); iii) cMPOpos/mCD33pos/mCD34neg/mHLA-DRneg to NPM1mut or acute promyelocytic leukemia exhibiting a t(15;17) (p<0.0001; sensitivity, 20%; specificity, 98%); iv) cMPOpos/cCD34pos/mCD34pos/mHLA-DRpos/mCD56neg/mCD7neg/mCD61neg/mCD41neg to inv(16) (p<0.0001; sensitivity, 75%; specificity, 91%). Conclusions: The present study shows strong associations between specific immunophenotype patterns and genotypes in AML, which might be useful for daily clinical practice and future genotype-specific treatment. Disclosures: Salih: Pfizer: Research Funding. Döhner:Pfizer: Research Funding. Schlenk:Celgene, Pfizer, Novartis, Amgen, Cephalon: Research Funding.

scholarly journals CD97 Is Associated with Poor Overall Survival in Acute Myeloid Leukemia

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 2794-2794
Author(s):  
Vijaya Pooja Vaikari ◽  
Sharon Wu ◽  
Houda Alachkar
Keyword(s):  
Overall Survival ◽  
Acute Myeloid Leukemia ◽  
Median Survival ◽  
Older Patients ◽  
Myeloid Leukemia ◽  
High Expression ◽  
Molecular Characteristics ◽  
Wild Type ◽  
Npm1 Mutation ◽  
Acute Myeloid

Abstract Introduction: Acute myeloid leukemia (AML) is a heterogeneous, hematologic malignancy characterized by clonal proliferation of myeloid precursors. Overall survival for patients with AML remains dismal (<50% for younger patients and <10% for older patients) due to high relapse rates, necessitating the identification of novel therapeutic targets. CD97 is a glycoprotein belonging to the EGF-TM7 molecule family, a subgroup of adhesion G-protein coupled receptors. CD97 was shown to play a role in cell migration, adhesion and interaction with cell surface proteins and components of the extracellular matrix. In several solid cancers, CD97 association with integrin was shown to regulate invasion, migration, angiogenesis, and poor survival outcome. Even though CD97 is normally expressed on myeloids and leukocytes, there is sparse data related to its expression on myeloid leukemia cells. In this study, we characterized CD97 expression in patients with AML using publically available data sets to determine its association with patient's clinical and molecular characteristics and clinical outcome. Methods: CD97 mRNA (RNA Seq V2 RSEM) expression and Z-score data was downloaded from TCGA. We generated Kaplan-Meier survival curves to compare overall (OS) and event-free (EFS) survival between patients with CD97 high (Z≥1) and CD97 low (Z<1) expression after stratification by age, cytogenetic status, transplant status and NPM1 mutation status. Association between CD97 expression and patient clinical and molecular characteristics was performed by Mann-Whitney U's non-parametric t-test and Fisher's exact test. STATA 12.0 SE was used to perform a Cox Proportional Hazards Model. Results: Patients with cytogenetically normal AML (CN-AML) had significantly higher CD97 expression than cytogenetically abnormal AML (CA-AML) (1.31 fold, p=0.023). Patients with high CD97 expression had significantly higher WBC count (median: 56.1 vs 13.1, p=0.004) and % bone marrow blasts (median: 83 vs 71%, p=0.019). CD97 was significantly higher in patients with NPM1 mutation (n=48) compared with patients with NPM1 wild type (n=125) (1.56 fold, p<0.000). CD97 was also significantly higher in patients with FLT3 mutation (ITD and point mutations) (n=49) compared with patients with FLT3 wild type (n=124) (1.4 fold, p=0.0008). Additionally, CD97 was significantly lower in the patients with RUNX1 mutation (n=17) compared with wildtype gene (n=156) (42.1% lower; p=0.0002). The OS of the CD97 high was significantly shorter than that of the CD97 low patients (median: 7.35 vs. 24.1 months; p=0.0015). Patients with CD97 high expression had significantly shorter EFS than that of CD97 low (median: 5.35 vs. 12 months; p=0.0015). Furthermore, in CN-AML CD97 high patients had shorter OS than CD97 low patients in CN-AML (median: 7.5 vs 20.5 months; p=0.0045; Figure 3B). In multivariate survival analysis, CD97 high expression was associated with shorter OS when adjusted for age, cytogenetic risk and transplant status (HR= 1.96; 95% Cl: 1.19-3.24; p=0.009). Because CD97 expression was associated with NPM1 mutation in AML, we stratified patients according to NPM1 mutational status and found that in patients with wildtype NPM1, CD97 high expression was associated with significantly shorter OS (Median survival: 6.35 vs 22.3 months; P= 0.0081) and EFS (Median survival: 5.35 vs 13.4 months; P=0.0006). In patients with NPM1 mutation, a similar but not significant association was observed with CD97 high expression and shorter OS (median survival: 7.5 vs 24.1 months; P=0.1) and EFS (median survival: 5.8 vs 11.1 months; P=0.27). Furthermore, in patients ≥ 60; CD97 high expression was associated with shorter OS compared with CD97 low (median survival 3.3 vs 11 months; p=0.0019) and EFS (Median survival 2.9 vs 9.2 months; p=0.0092). Similar but not significant association was observed in in patients<60 (Median OS: 27.5 vs 53.9; p=0.2) and (Median EFS 8.5 vs 53.9; p=0.2). Conclusion: Patients with high CD97 expression had shorter OS and EFS. CD97 was higher in CN-AML compared with CA-AML patients. Additionally, high CD97 was also associated with NPM1 mutation. Our findings demonstrate that CD97 expression contributes to the clinical outcome of patients with AML, particularly in older patients. This study provides rationale for further functional and mechanistic studies aiming to understand the role of CD97 in AML. Disclosures No relevant conflicts of interest to declare.

Association of TET2 Alterations with NPM1 Mutations and Prognostic Value in De Novo Acute Myeloid Leukemia (AML).

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 163-163 ◽  
Author(s):  
Olivier Nibourel ◽  
Olivier Kosmider ◽  
Meyling Cheok ◽  
Nicolas Boissel ◽  
Aline Renneville ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Research Funding ◽  
Prognostic Value ◽  
De Novo ◽  
Missense Mutations ◽  
Npm1 Mutation ◽  
Tet2 Mutation ◽  
De Novo Aml ◽  
Acute Myeloid

Abstract Abstract 163 In acute myeloid leukemia (AML), both cytogenetic and molecular abnormalities are strongly associated with prognosis. In particular, in cytogenetically normal AML (CN-AML), FLT3-ITD (internal tandem duplication) carries adverse prognostic factor whereas NPM1 or CEBPA mutations are associated with favorable outcome. Recently, mutations of the ten eleven translocation 2 gene (TET2) have been reported myeloid neoplasms. We evaluated the frequency and prognostic value of TET2 alterations, in a cohort of 111 de novo AML patients. We studied 111 patients aged between 15 years and 69 years with previously untreated de novo AML who had reached complete remission (CR) using intensive chemotherapy. 28 of them also received an allogenic bone marrow transplantation in first CR. Analysis of TET2 sequence variation was performed by direct sequencing of PCR products from 111 genomic DNA samples obtained at diagnosis. Frameshift and nonsense variations were all scored as mutation whereas missense mutations were retained when observed at diagnostic but absent in the CR paired sample obtained. Previously identified single nucleotide polymorphisms (SNP) were not considered. TET2 anomalies were numbered according to Genebank reference FM992369. Paired diagnosis and CR genomic DNAs were analyzed using Affymetrix Genome-Wide Human SNP Array 6.0 (Affymetrix, Santa Clara, CA). Data were analyzed using Gene Chip Genotyping Console 3.0.2 and Partek Genomics Suite (www.partek.com/). Comparisons were made by Fisher's exact test for binary variables and the Mann-Whitney‘s test for continuous variables. Disease Free Survival (DFS) and overall survival (OS) were calculated according to the Kaplan-Meier method. Comparisons regarding DFS and OS were performed with the log-rank test. 24 acquired TET2 mutations were observed in 19 of the 111 (17%) de novo AML patients, suggesting the alteration of the two TET2 alleles in 5 patients. They included 21 different events: 6 frameshift, 7 non-sense and 11 missense mutations. Four of the missense mutations were located in conserved regions and 7 outside. All of them were detected in the diagnostic sample but were absent in the paired remission sample. Except for two missense mutations (S282F, T492S) both detected in two patients, no recurrent TET2 mutation was observed. Acquired mutations were spread over all exons. No case of uniparental disomy (UPD) was observed and only one patient presented a small deletion of 60Kb in the TET2 gene locus without TET2 mutation. No significant difference was observed between patients with or without TET2 alterations for gender, age, hemoglobin level, platelet count, FAB subtypes distribution and cytogenetics according to MRC classification, but there was a trend for higher WBC count in patients with TET2 alteration. No significant association was observed between TET2 mutations and FLT3 or CEBPA alterations. However, TET2 alterations were significantly associated with NPM1 mutations (p=0.032). In the entire patient cohort, no difference in DFS or OS was seen between patients with and without TET2 alteration. However, a significantly worse DFS was observed for patients presenting TET2 mutations within the subgroup of patients with NPM1 mutations (3y-DFS: 0% vs 66.4%, 95% CI [45.6–87.2], p=0.008) Considering both the favorable prognosis of NPM1 mutations without FLT3-ITD in CN-AML and the absence of clear association between FLT3-ITD and TET2 alterations in this study, prognostic value of the genotype characterized by NPM1 mutation without FLT3-ITD or TET2 alteration (NPM1+FLT3-ITD-TET2-) was compared to other patients within CN-AML group (N=54). NPM1+FLT3-ITD-TET2- patients showed a significantly better DFS and OS compared to other patients in CN-AML group (3y-DFS: 82.1%, 95% CI [59.1–100] vs 37.3%, 95% CI [20.2–54.3], p=0.01; 3y-OS: 80.8%, 95% CI [56.1–100] vs 42.3%, 95% CI [23.3–61.3], p=0.04). In conclusion, we observed point mutations of TET2 in 17% of patients, whereas TET2 deletion or UPD are very rare. In our study, TET2 mutations were clearly associated with NPM1 mutations and carried a negative prognostic impact in this subgroup. Screening for TET2 mutations may improve the characterization of CN-AML and help to identify within the low-risk subgroup with NPM1 mutation and without FLT3-ITD, patients at high risk of relapse. Disclosures: Fenaux: Celgene: Honoraria, Research Funding; Roche: Honoraria, Research Funding; Ortho Biotech: Honoraria, Research Funding; Amgen: Honoraria, Research Funding; Cephalon: Honoraria, Research Funding; Merck: Honoraria, Research Funding; Novartis: Honoraria, Research Funding.

In Acute Myeloid Leukemia, the Use in Induction of Standard Dose Arac Is Associated with a Better Quality of Response as Compared to An Induction Regimen Containing High Dose Arac.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1584-1584
Author(s):  
Luca Maurillo ◽  
Francesco Buccisano ◽  
Alessandra Spagnoli ◽  
Maria Ilaria Del Principe ◽  
Benedetta Neri ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Standard Dose ◽  
Lower Probability ◽  
High Dose ◽  
Npm1 Mutation ◽  
Induction Regimen ◽  
The Impact ◽  
Acute Myeloid

Abstract Abstract 1584 Poster Board I-610 The clinical advantage of using high-dose cytarabine (HDARAc) in induction chemotherapy for acute myeloid leukemia (AML) is still controversial. The purpose of our study was to explore the impact on the “quality of response” of an induction regimen containing standard dose (SD) ARAc versus HDARAc, by measuring minimal residual disease (MRD) once CR was achieved. MRD was determined by multiparametric flow cytometry on bone marrow samples collected at the end of induction and consolidation therapy. The threshold for MRD negativity was set below the number of 3.5×10-4 residual leukemic cells. We evaluated 111 patients with de novo AML, enrolled sequentially in DCE arm of AML10 (n=40) and in AML12 (n=71) EORTC/GIMEMA randomized trials between 1995 and 2007. In DCE arm of AML10, induction treatment combined ARAc (100 mg/m2 day 1-10), etoposide (50 mg/m2 day 1-5), and on days 1,3,5, daunorubicin (50 mg/m2). In AML12 trial, patients received the same treatment of DCE arm except for ARAc dose that was 100 mg/m2 day 1-10 or 3000 mg/m2/q12 hrs on days 1,3,5, and 7 according to randomization. As consolidation, all patients received ARAc (500 mg/m2/q12 hrs day 1-6) and daunorubicin (50 mg/m2, day 4-6). Median age was 45 yrs (range 18-60), 65 males and 46 females. Seventy-five patients were treated on SDARAc regimen and 36 on HDARAc regimen. The two groups were well balanced in terms of FAB classes, WBC count, cytogenetics, Pgp-170 expression, FLT3 and NPM1 mutation and post-remission transplant procedure (autologous or allogeneic) delivered. After induction, we observed a significantly higher frequency of MRD negativity in SDARAc arm vs HDARAc arm (82.% vs 18%, p=0.02). After consolidation, this figure was confirmed (84% vs 16%, p=0.01). At this stage, 3 further patients treated on the SDARAc arm, became MRD negative, whereas none in the HDARAc arm did so. Overall, median level of MRD was significantly lower in SDARAc group both after induction (1.1×10-2 vs 5.4×10-2, p=0.015) and consolidation (9×10-3 vs 3×10-2, p=0.02). Based on the combination of MRD status after consolidation and ARA-C schedule delivered, we identified 4 different groups of patients. Five years OS for SDARAc-MRDneg, HDARAc-MRDneg, SDARAc-MRDpos and HDARAc-MRDpos was 67%, 33%, 21% and 24%, respectively (p<0.0001). Similarly, 5 years DFS for SDARAc-MRDneg, HDARAc-MRDneg, SDARAc-MRDpos and HDARAc-MRDpos was 67%, 33%, 13% and 26%, respectively (p<0.0001). Since calculation of survival estimates is influenced by the occurrence of toxic events, 3 patients in the HDARAc-MRDneg group died because of complications, cumulative incidence of relapse (CIR) was also evaluated. Five years CIR for SDARAc-MRDneg, HDARAc-MRDneg, SDARAc-MRDpos and HDARAc-MRDpos was 20% (95% CI, 19-21), 17% (95% CI, 12-24), 75%(95% CI, 74-76) and 57% (95% CI, 55-60), respectively (p<0.0001); analysis of CIR confirmed the prognostic importance to achieve a MRD negative status at the end of consolidation, whatever the regimen used. In conclusion, delivery of an induction regimen containing daunorubicin, etoposide and SDARAc given for 10 days, results in a more efficient clearance of leukemic burden compared to a similar regimen HDARAc based. Such superior efficiency translates into a better “quality” of response, as demonstrated by the more frequent achievement of a MRD negative status, and then into a more favorable outcome. In addition, it should also be considered that the lower probability to gain a condition of MRD negativity by giving HDARAc, might further be jeopardized by fatal toxicities. Disclosures No relevant conflicts of interest to declare.

Incidence and Prognostic Value of FLT-3 and NPM1 Genes Abnormalities in Acute Myeloid Leukemia in the Côte D’or Population, France

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4873-4873
Author(s):  
Marc Maynadié ◽  
Martine Courtois ◽  
Morgane Mounier ◽  
Ines Janoray-Manivet ◽  
Ingrid Lafon ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Prognostic Value ◽  
De Novo ◽  
Relative Survival ◽  
Normal Karyotype ◽  
Npm1 Mutation ◽  
Clinical Series ◽  
De Novo Aml ◽  
Acute Myeloid

Abstract Context: In acute myeloid leukemia (AML), the recently described FLT-3 and NPM1 genes abnormalities were found to have a prognostic value in AML with normal karyotype and a specific therapeutic strategy was proposed according to these abnormalities. We look for the incidence and prognostic value of these abnormalities in cases diagnosed on a well defined population. Material and Methods: AML diagnosed according to WHO classification between 01/01/2001 and 31/12/2006 in the population of the Côte d’Or department, were included. Karyotype analyses were performed in 81% of the cases. The FLT3 D835 mutation, the FLT3 internal duplication (ITD) and the NPM1 mutation were systematically studied on the biological material of the diagnosis cryopreserved in the Ferdinand Cabanne Biobank of Burgundy, by PCR and DNA sequencing techniques. The vital status of the patients was update on 31/10/2007. The relative survival was calculated with the STATA (V9) software. Results: 100 de novo AML and 47 secondary AML (sAML) were registered (72 females and 75 males). The world standardized incidence rate was 2.4 in men and 1.5 in women for de novo AML instead of what it was respectively of 1.1 and 0.6 in sAML. The urban predominance was present in both type of AML. The karyotype was normal in 38% (45/119) of cases (35% of de novo AML and 21% of sAML). It was abnormal in 62% of cases (74/119)(51% of de novo AML and 49% of sAML). Molecular analyses were performed in 78 de novo AML and in 24 sAML. FLT3 ITD was present in 19% (15/78) de novo AMl and in any sAML. The FLT3 D835 mutation was present in 6.5% of de novo AML and in 8% of sAML. NPM1 was mutated respectively in 26% and 4% of the cases. There was a significantly higher WBC count and proportion of blast cells in peripheral blood in FLT3 ITD cases. Overall and relative survivals of FLT3 ITD cases were decreased compared to FLT3 wild type cases. No difference according to NPM1 status was found. Conclusions: These data confirm the bad prognostic value of FLT3 ITD status in AML observed in clinical series. Furthermore their particular interest lies in the fact that they are the first molecular data in AML produced on a population-based series indicating the feasibility of such epidemiological studies.

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