Normal karyotype acute myeloid leukemia with the CD7+ CD15+ CD34+ HLA-DR + immunophenotype is a clinically distinct entity with a favorable outcome

Abstract The transcription factor CCAAT/enhancer binding protein alpha (C/EBPa) encoded by the CEBPA gene, is crucial for the differentiation of immature granulocytes. Diminished or abnormal C/EBPa activity resulting from CEBPA gene mutations is widely known to contribute to the transformation of myeloid progenitors via reduction of their differentiation potential. The CEBPA mutations have been detected in approximately 7% of total acute myeloid leukemia (AML) and in 15% of those with intermediate-risk cytogenetics or those with normal karyotype. However, the age distribution of the patients with the CEBPA mutations and the immunophenotype of their leukemic cells are not known. Sequential studies of the CEBPA gene in AML patients are also limited. In this study, 104 patients with de novo acute myeloid leukemia (AML) were evaluated for the CEBPA mutation by direct sequencing. Excluding the silent mutations, 16 (15%) of the total 104 AML patients, 15 (25%) of the 61 patients with intermediate-risk cytogenetics and 11 (35%) of the 31 patients with normal karyotype showed CEBPA mutations, frequencies higher than those reported in the West. Further cloning and subsequent nucleotide sequence analysis revealed that 14 patients had heterozygous biallelic mutations: 11 had mutations involving both the N-terminal transactivation domain (TAD) and the C-terminal basic leucine zipper domain (bZIP) and three, in either the TAD region or the bZIP region. The remaining two patients had only one allele mutation in the TAD1 region. Most mutations in TAD region were repeat-number changes of simple sequence repeats and those in bZIP region were internal tandem duplications. Sequence analysis revealed that in the region spanning the bZIP mutations, there was hot spot for concensus topoisomerase II sites, which has also been shown in other AML-related mutations FLT3-ITD and MLL duplication. All but one patient with CEBPA mutations had M1 or M2 subtype of AML. The patients with CEBPA mutations had significantly higher incidences of CD7 (73%), CD15 (100%), CD34 (93%) and HLA-DR (93%) expression than others and the majority of them showed a distinct immunophenotype of the leukemic cells: HLA-DR+ CD7+ CD13+ CD14− CD15+ CD33+ CD34+. The incidence of the CEBPA mutation in children with AML was similar to that in adults. The CEBPA mutation was serially analyzed in 27 patients; the mutations disappeared at CR, but reappeared at relapse. No one developed novel mutation during the follow-up period. In conclusion, the CEBPA mutation may play an important role in the development, but not progression, of AML. The patients with the CEBPA mutations showed a distinct immunophenotype of the leukemic cells. Potential topoisomerase II cleavage sites locating in the bZIP region were first reported and we propose that this is relevant to the process of illegitimate recombination generating the internal tandem duplication pattern of bZIP mutations.

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Correlation of Next-Generation Sequencing Based Genotypic Profiles with MICM Characteristics in NPM1 Mutated Acute Myeloid Leukemia

10.21203/rs.3.rs-41514/v1 ◽

2020 ◽

Author(s):

Biao Wang ◽

Bin Yang ◽

Wei Wu ◽

Xuan Liu ◽

Haiqian Li

Keyword(s):

Acute Myeloid Leukemia ◽

Next Generation Sequencing ◽

Myeloid Leukemia ◽

Normal Karyotype ◽

Next Generation ◽

Ras Pathway ◽

Logistic Analysis ◽

Hla Dr ◽

Generation Sequencing ◽

Acute Myeloid

Abstract The purpose of this study was to analyze association between next-generation sequencing (NGS) genomic profile and conventional MICM characteristics in patients with acute myeloid leukemia (AML) with NPM1 mutation (NPM1mut). We selected 238 NPM1mut patients with available NGS information on 112 genes related to blood diseases, using χ2 and Mann-Whitney U test to analyze the distribution correlation between genomic alterations and MICM parameters. Compared with NPM1mut/FLT3-ITD(−), the NPM1mut/FLT3-ITD(+) group presented a slightly common M5 [78/143 (54.5%) vs. 64/95 (67.4%); P=0.048], more higher CD34 and CD7 expression rates (CD34: 20.6% vs. 47.9%, P<0.001; CD7: 29.9% vs. 61.5%, P<0.001), and lack of favorable- and adverse-risk karyotypes (6.4% vs. 0%; P=0.031). In entire NPM1mut cohort, totaling 240 NPM1 mutation events were identified, of whom 10 (10/240, 4.2%) were missense types. When confining analysis to the 205 cases with NPM1mut insertions/deletions type and normal karyotype, multivariable logistic analysis showed that FLT3-ITD was positively correlated with CD34 and CD7 expression (HR=5.29 [95% CI 2.64-10.60], P<0.001; HR=3.47 [95% CI 1.79-6.73], P<0.001; respectively). Ras-pathway mutation was positively correlated with HLA-DR expression (HR=4.05 [95% CI 1.70-9.63], P=0.002), and KRAS mutation negatively with MPO expression (HR=0.18 [95% CI 0.05-0.62], P=0.007). DNMT3A-R882 was positively correlated with CD7 and HLA-DR expression (HR=3.59 [95% CI 1.80-7.16], P<0.001; HR=13.41 [95% CI 4.56-39.45], P<0.001; respectively). DNMT3A mutation was negatively correlated with MPO expression (HR=0.35 [95% CI 1.48-8.38], P=0.004). TET2/IDH1 mutations were negatively correlated with CD34 and CD7 expression (HR=0.26 [95% CI 0.11-0.62], P=0.002; HR=0.30 [95% CI 0.14-0.62], P=0.001; respectively), and positively with MPO expression (HR=3.52 [95% CI 1.48-8.38], P=0.004). NPM1mut co-existing mutations in signaling pathways (FLT3-ITD and Ras-pathway) and methylation modifiers (DNMT3A and TET2/IDH1) are linked with the expression of CD34, CD7, HLA-DR and MPO, thereby providing a mechanism explanation for the immunophenotypic heterogeneity of this AML entity.

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Favorable outcome of patients with normal karyotype acute myeloid leukemia harboring FLT3-ITD and treated with cladribine added to daunorubicin and cytarabine induction

Acta Haematologica Polonica ◽

10.1016/j.achaem.2015.07.048 ◽

2015 ◽

Vol 46 ◽

pp. 27

Author(s):

M. Libura ◽

S. Giebel ◽

B. Piątkowska-Jakubas ◽

M. Przestrzelska Pawełczyk ◽

I. Florek ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Normal Karyotype ◽

Favorable Outcome ◽

Acute Myeloid

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Molecular and cytogenetic abnormalities in acute myeloid leukemia: review and case studies

Einstein (São Paulo) ◽

10.1590/s1679-45082011ao2041 ◽

2011 ◽

Vol 9 (2) ◽

pp. 184-189 ◽

Cited By ~ 2

Author(s):

Elvira Deolinda Rodrigues Pereira Velloso ◽

Carlos Henrique Ares Silveira da Motta ◽

Juliana Braga Furtado ◽

Nydia Strachman Bacal ◽

Paulo Augusto Achucarro Silveira ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Normal Karyotype ◽

Good Prognosis ◽

Correct Classification ◽

Npm1 Mutation ◽

Small Series ◽

Hla Dr ◽

Acute Myeloid

Objective: To study the frequency of mutations that may lead to a good or bad prognosis, as well as their relation with the karyotype and immunophenotype in patients with acute myeloid leukemia. Methods: Thirty samples of patients with acute myeloid leukemia were studied, in which FLT3-ITD, FLT3-TKD and NPM1 mutations were investigated. All samples were submitted to immunophenotyping and 25 to karyotyping. Results: An occurrence of 33.3% NPM1 mutation and an equal number of FLT3-ITD mutation were observed. When only the cases with normal karyotype were studied, this figures increased to 50 and 40%, respectively. Eight percent of cases with normal karyotype and genotype NPM1+/FLT3- were included in the group of acute myeloid leukemia with good prognosis. The typical phenotype of acute myeloid leukemia with normal karyotype and mutated NPM1 (HLA-DR and CD34 negative) was not observed in this small series. Conclusion: Good prognosis cases were identified in this series, emphasizing the need to include new genetic markers in the diagnostic routine for the correct classification of acute myeloid leukemia, to more properly estimate prognosis and determine treatment.

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The favorable impact of CEBPA mutations in patients with acute myeloid leukemia is only observed in the absence of associated cytogenetic abnormalities and FLT3 internal duplication

Blood ◽

10.1182/blood-2008-12-194704 ◽

2009 ◽

Vol 113 (21) ◽

pp. 5090-5093 ◽

Cited By ~ 58

Author(s):

Aline Renneville ◽

Nicolas Boissel ◽

Nathalie Gachard ◽

Dina Naguib ◽

Christian Bastard ◽

...

Keyword(s):

Overall Survival ◽

Acute Myeloid Leukemia ◽

Confidence Interval ◽

Myeloid Leukemia ◽

Normal Karyotype ◽

Favorable Outcome ◽

Abnormal Karyotype ◽

Cytogenetic Abnormalities ◽

The Impact ◽

Acute Myeloid

AbstractMutations of the CCAAT/enhancer binding protein alpha (CEBPA) gene have been associated with a favorable outcome in patients with acute myeloid leukemia (AML), but mainly in those with a normal karyotype. Here, we analyzed the impact of associated cytogenetic abnormalities or bad-prognosis fms-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD) in 53 patients with CEBPA+ de novo AML treated in the Acute Leukemia French Association trials. We found that only those with a normal karyotype and no FLT3-ITD displayed the expected favorable outcome. In this context, relapse-free, disease-free, and overall survival were significantly longer than in corresponding patients without the CEBPA mutation (P = .035, .016, and .047, respectively). This was not observed in the context of an abnormal karyotype or associated FLT3-ITD. Furthermore, after adjustment on age, trial, and mutation type, these features were independently predictive of shorter overall survival in the subset of patients with CEBPA+ AML (multivariate hazard ratio = 2.7; 95% confidence interval, 1.08-6.7; and 2.9; 95% confidence interval, 1.01-8.2; with P = .034 and .05, for abnormal karyotype and FLT3-ITD, respectively).

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FLT3 Internal Tandem Duplication in Acute Myeloid Leukemia with Normal Karyotype

The Korean Journal of Hematology ◽

10.5045/kjh.2007.42.3.250 ◽

2007 ◽

Vol 42 (3) ◽

pp. 250 ◽

Cited By ~ 1

Author(s):

Sang-Ho Kim ◽

Yeo-Kyeoung Kim ◽

Il-Kwon Lee ◽

Deog-Yeon Jo ◽

Jong-Ho Won ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Tandem Duplication ◽

Normal Karyotype ◽

Internal Tandem Duplication ◽

Flt3 Internal Tandem Duplication ◽

Acute Myeloid

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FLT3 mutations have no prognostic impact in elderly patients with acute myeloid leukemia and normal karyotype

American Journal of Hematology ◽

10.1002/ajh.21458 ◽

2009 ◽

Vol 84 (8) ◽

pp. 532-534 ◽

Cited By ~ 11

Author(s):

Felicetto Ferrara ◽

Clelia Criscuolo ◽

Cira Riccardi ◽

Tiziana Izzo ◽

Mariangela Pedata ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Elderly Patients ◽

Myeloid Leukemia ◽

Normal Karyotype ◽

Prognostic Impact ◽

Flt3 Mutations ◽

Acute Myeloid

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BCOR Mutations in Acute Myeloid Leukemia: Clonal Evolution and Prognosis

Blood ◽

10.1182/blood-2020-137127 ◽

2020 ◽

Vol 136 (Supplement 1) ◽

pp. 4-4

Author(s):

Ashley Zhang ◽

Yuntao Liu ◽

Shuning Wei ◽

Benfa Gong ◽

Chunlin Zhou ◽

...

Keyword(s):

Bone Marrow ◽

Acute Myeloid Leukemia ◽

Risk Stratification ◽

Myeloid Leukemia ◽

Statistical Difference ◽

Tp53 Mutation ◽

Clonal Evolution ◽

Normal Karyotype ◽

Abnormal Karyotype ◽

Acute Myeloid

Background BCOR gene is a transcription repressor that may influence normal hematopoiesis and is associated with poor prognosis in acute myeloid leukemia (AML) with normal karyotype. However, due to the rare mutation frequency in AML (3.8%-5%), clinical characteristics and prognosis of AML patients with BCOR mutation including abnormal karyotype are still unknown. In addition, the clonal evolution of AML patients with BCOR mutation has not been fully investigated. Methods By means of next generation of sequencing, we performed sequencing of 114 genes related to hematological diseases including BCOR on 509 newly diagnosed AML patients (except for acute promyelocytic leukemia) from March 2017 to April 2019. The 2017 European Leukemia Net (ELN) genetic risk stratification was used to evaluate prognosis. Overall survival (OS) was defined as the time from diagnosis to death or last follow-up. Relapse-free survival (RFS) was measured from remission to relapse or death. Clonal evolution was investigated through analyzing bone marrow samples at diagnosis, complete remission (CR) and relapse from the same patient. Result Among 509 AML patients, we found BCOR mutations in 23 patients (4.5%). BCOR mutations were enriched in patients with mutations of RUNX1 (p = 0.008) and BCORL1 (p = 0.0003). Patients with BCOR mutation were more at adverse ELN risk category compared to patients without BCOR mutation (p = 0.007). Besides, there was a larger proportion of patients with normal karyotype in BCOR mutation group but it had not reached statistical difference (62.5% vs 45.5%, p = 0.064). The abnormal karyotype in patients with BCOR mutations included trisomy 8, t(9;11), inv(3), -7 and complex karyotype.There were no significant differences in age, sex, white blood cell count, hemoglobin or platelet count between the two groups. More patients died during induction (13.0% vs 3.5%, p = 0.56) and fewer patients achieved CR after 2 cycles of chemotherapy when patients had BCOR mutations (69.6% vs 82.5%, p = 0.115) but the difference had not reached statistical difference . Patients with BCOR mutations had inferior 2-year OS (52.1% vs 70.7%, p = 0.0094) and 2-year RFS (29.8% vs 61.1%, p = 0.0090). After adjustment for ELN risk stratification, BCOR mutation was still remain a poor prognostic factor. However, the adverse prognostic impact of BCOR mutation is overcome by hematopoietic stem cell transplantation (HSCT), in which there was no difference between BCOR mutation group and wild type group (p = 0.474) (Figure 1). Through analysis of paired bone marrow sample at diagnosis, remission and relapse, we revealed the clonal evolution that BCOR mutation was only detected at diagnosis sample as a subclone and diminished at CR and relapse while TP53 mutation was only detected at relapse with a variant allele frequency (VAF) of 25.5%. We also found BCOR mutation at another patient's diagnosis and relapse sample while TP53 mutation was detected at relapse with VAF of 11.8%. Conclusion BCOR is associated with RUNX1 mutation and higher ELN risk. AML patients with BCOR mutation including normal and abnormal karyotype conferred a worse impact on OS that can be overcome by HSCT. BCOR mutation is a subclone at diagnosis or relapse in some patients, in which TP53 mutation clone occurred at relapse. Disclosures No relevant conflicts of interest to declare.

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