1.The clinical characteristics of newly diagnosed acute myeloid leukemia patients with NPM1 mutation.
Hong-Hu ZHU ; Bin JIANG ; Xi-Jing LU ; Qian JIANG ; Hao JIANG ; Li BAO ; Xiao-Hui ZHANG ; Jin LU ; Guo-Rui RUAN ; Yue-Yun LAI ; Yan-Rong LIU ; Xiao-Jun HUANG
Chinese Journal of Hematology 2010;31(5):315-318
OBJECTIVETo investigate the clinical characteristics of newly diagnosed acute myeloid leukemia (AML) with NPM1 mutation.
METHODSNPM1 mutation (including A, B, D mutation type) was detected in 206 patients with newly diagnosed AML by real-time quantitative RT-PCR.
RESULTSThe incidence of NPM1 mutation was 15.5% in total AML patients and 32.5% in normal karyotypes AML patients. The characteristics of 174 NPM1 wild type patients v.s. that of 32 NPM1 mutation patients was as follow, median age (46 vs 35 years old, P < 0.01), WBC counts (27 × 10(9)/L vs 8 × 10(9)/L, P < 0.01), BPC (82 × 10(9)/L vs 36 × 10(9)/L, P < 0.01), proportion of AML-M(5) (31.2% vs 5.8%, P = 0.01), incidence of normal karyotypes (92.6% vs 40.8%, P < 0.01), incidence of FLT3-ITD-positive (25.0% vs 7.5%, P < 0.01), CD34-positvie (23.3% vs 69.5%, P < 0.01), cases with fusion gene (0 vs 47.1%, P < 0.01). No statistic difference was found in sex, percentage of blasts in bone marrow, complete remission rate, overall survival between the two groups. Relapse-free survival in AML patients with NPM1-mutation and FLT3-ITD-negative tended to be higher than in those with NPM1-mutation and FLT3-ITD-positive.
CONCLUSIONIt is necessary to detect NPM1 mutation and FLT3-ITD in newly diagnosed AML patients, especially in patients with high WBC and BPC, CD34-negative, normal karyotype, which might help to molecular classification and treatment.
Humans ; Leukemia, Myeloid, Acute ; genetics ; Mutation ; Nuclear Proteins ; genetics ; Prognosis ; fms-Like Tyrosine Kinase 3 ; genetics
2.ITD mutation burden for the prognosis in FLT3-ITD positive acute myeloid leukemia patients.
Shasha DING ; Hongjie SHEN ; Zixing CHEN ; Suning CHEN ; Jiannong CEN ; Zixuan DING ; Jun HE
Chinese Journal of Hematology 2015;36(6):449-454
OBJECTIVETo explore the impact of ITD mutation characteristics on the overall survival (OS) and complete remission duration (CRD) in FLT3-ITD positive non-M3 acute myeloid leukemia (AML).
METHODSCapillary electrophoresis was used to detect the FLT3-ITD characteristics after PCR amplication. Single or multiple mutations were identified by the numbers of peak. FLT3-ITD mutation burden was calculated by the peak area of mutant divided by the wild-type and mutant peak areas. Clinical data was collected and followed up in the FLT3-ITD mutation patients.
RESULTSMultiple ITD mutations were common in patients aged 60 and above. Patients with single ITD mutation had higher percentage of blasts in bone marrow than multiple ITD mutations (0.758 vs 0.638, P=0.028). The numbers and length of FLT3-ITD mutation had no impact on prognosis. Patients with less than 10% of ITD mutation burden showed no difference with the intermediate-risk c-kit group in OS and CRD, but the two groups had longer OS and CRD than ITD mutation burden above 10% (OS: undefined, undefined, 9.9 months, P<0.05; CRD: undefined, undefined, 6.7 months, P<0.05). In patients with ITD mutation burden above 10%, cases with NPM1 or CEBPA mutation alone had markedly longer CRD than ITD mutation alone (25.0 vs 5.1 months, P=0.003), while OS were similar (11.4 vs 8.0 months, P>0.05).
CONCLUSIONNon-M3 AML patients with less than 10% FLT3-ITD mutation burden had a better prognosis than those above 10%.
Genotype ; Humans ; Leukemia, Myeloid, Acute ; Mutation ; Prognosis ; Remission Induction ; fms-Like Tyrosine Kinase 3
4.Analysis of Clinical and Laboratory Features of Patients with Acute Myeloid Leukemia(AML)-M1/M2 with Cuplike Nuclei Morphology.
Yan WANG ; Chun QIAO ; Rui GUO ; Hui WANG ; Hui YANG ; Hai-Rong QIU ; Yu-Jie WU ; Jian-Fu ZHANG ; Rong WANG ; Si-Xuan QIAN ; Jian-Yong LI
Journal of Experimental Hematology 2018;26(4):958-963
OBJECTIVETo analyze the clinical and laboratory features of acute myeloid leukemia (AML) with cuplike nuclei morphology.
METHODSOne hundred and seventy patients diagnosed with AML (M1andM2) between December 2009 and December 2016 were included in the study. Bone marrow smears were prepared for morphologic alanalysis, the immunophenotype was analyzed by flow cytometry and the RHG-banding was for conventional cytogenetic assay (CCA) ,gene mutation was detected by sequencing.
RESULTSAmong the 170 AMLpatients, 67 were diagnosed as M1 and 103 patients was diagnosed as M2, 43 patients(25.3%) defined as cuplike nuclei-positive, among them 38patients (88.4%) were M1 while only 5 patients (11.6%) were with M2(P<0.01). No significant value about sex(P> 0.05) between cuplike nuclei-positive and -negative group, while older patients were found in cuplike nuclei-positive group (P<0.05). Higher peroxydas (POX) ratio (P<0.05) and integration (P<0.05) were found in cuplike nuclei- positive group. Furthermore, the patients with cuplike nuclei-positive lack the expressions of CD34 (P<0.01) and HLA-DR(P<0.01) while no other immunophenotype markers were found. Among the 152 patients (89.4%) for genetic analysis ,83.8% karyotype of the cuplike nuclei-positive group were normal while only 54.8 of negative group was normal by CCA. Molecular biology analysis showed that the patients in cuplike nuclei-positive group have significantly highe rNMP1 (P<0.01) and FLT3(P<0.01) mutations as compared with the negative group. Furthermore, the relationship of the ratio o fcuplike nuclei and the type of gene mutations were investigated, and no significant associations were found. However, it was found that the patients with FLT3 mutation displayed more biological nuclear invagination than the patients with NPM1 mutations (P<0/01).
CONCLUSIONAML patients with positive cuplike nuclei have characteristic morphological changes, typical immunophenotype with HLA-DR- and CD34, normal karyotype accompanied by NPM1 and FLT3 mutations.
Cell Nucleus ; Humans ; Leukemia, Myeloid, Acute ; Mutation ; Nuclear Proteins ; Prognosis ; fms-Like Tyrosine Kinase 3
5.Clinical Characteristics and Prognosis of Acute Myeloid Leukemia with FLT3-ITD Mutation.
Yan LIU ; Xiao-Yan KE ; Jing WANG ; Ji-Jun WANG ; Hong-Mei JING ; Fei DONG
Journal of Experimental Hematology 2018;26(2):354-358
OBJECTIVETo investigate the clinical characteristics of acute myeloid leukemia(AML) patients with FLT3-ITD(Fms-like tyrosine kinase3, intenal tandem duplication) mutation and their response to treatment.
METHODSRetrospective analysis of 128 newly diagnosed AML (except type M3) patients was performed between January 2014 and July 2017. Patients were divided into FLT3-ITD mutated group and non-mutated group. Mutation detection was carried out by using polymerase chain reaction(PCR) and gene sequencing analysis. Standard 3 + 7 or CAG regimen were taken as the first induction chemotherapy, 4 cases received sorafenib, overall survival (OS) was calculated by Kaplan-Meier.
RESULTSNinety-seven patients can be evaluable for clinical data available; 4 patients were FLT3-TKD mutated, which accounted for 4.1%; 19 patients were FLT3-ITD mutated, which accounted for 19.59%(19/97). Median white blood cell count (WBC), percentage of peripheral blasts and LDH value were significantly higher in FLT3-ITD group than those in non-mutated group [64.65(1.07-587.92)×10/L vs 39.68 (0.45-203.81) ×10/L](P<0.01), [69.62(16-99)% vs 36.35(0-92) %](P<0.01 ) and [LDH 526(124-2 729)U/L vs 265(20-1977)U/L](P<0.05), respectively. The frequency of coexisting NPM1 mutation was higher in FLT3-ITD group than that in non-mutated group [36.8(7/19)% vs 6.8 (5/74) %](P<0.01). The CR+PR was lower in FLT3-ITD group than that in non-mutated group [31.6(6/19)% vs 64.9 (48/74)%](P<0.05). OS in FLT3-ITD group was significantly shorter than that in non-mutated group (5 vs 18 months)(P<0.05). There is no significant difference in OS between FLT3-ITD concomitant with and without NPM1 mutation groups(5 vs 5 months)(P>0.05). The median OS was 13 months for the FLT3-ITD patients taking sorafenib.
CONCLUSIONThe FLT3-ITD is a common mutation in AML, FLT3-ITD mutated AML is more likely concomitant with NPM1 mutation with higher number of WBC, percentage of peripheral blasts and LDH value, thus CR is low after the 1st treatment and survival is poor.
Humans ; Leukemia, Myeloid, Acute ; Mutation ; Prognosis ; Retrospective Studies ; Tandem Repeat Sequences ; fms-Like Tyrosine Kinase 3
6.NPM1 High Mutant Allele Burden is an Adverse Prognostic Factor for AML Patients with Mutated NPM1.
Jiang-Xue HOU ; Shu-Juan WANG ; Yan-Fang LIU ; Qian-Qian HAO ; Chong WANG ; Tao LI ; Jun-Jun BAI ; Lin-Xiao LIAO ; Cheng-Yu GUO ; Yin-Yin CHANG ; Meng WANG ; Hui SUN ; Xin-Sheng XIE ; Zhong-Xing JIANG
Journal of Experimental Hematology 2019;27(2):365-372
OBJECTIVE:
To investigate the clinical features, accompanying gene mutation characteristics and prognostic factors of adult patients with acute myeloid leukemia with mutated NPM1 (NPM1AML).
METHODS:
Seventy-three patients with newly diagnosed adult NPM1AML were selected. The mutations of 22 genes were detected by second generation sequencing and 43 fusion genes of AML were detected by real-time fluorescent quantitative PCR. The Kaplan-Meier survival curve and Cox multivariate regression analysis were used to study the prognostic factors.
RESULTS:
A total of 74 NPM1 site mutations were detected in 73 patients with NPM1AML. The incidence rates were 92.0% L287fs, 2.7% Q289fs and W288fs, 1.4% L258fs and Q289H, among which 1 patient had 2 NPM1 mutations; the different mutation sites had no effect on the prognosis of NPM1AML. The median value of NPM1 variant allele frequency (VAF) was 35.4% (1.8%-56.6%). Based on the uppermost quartile of 38.4%, the patients were classified as NPM1 VAF>38.4% (NPM1AML) and NPM1 VAF≤38.4% (NPM1AML). Compared with NPM1AML, the early mortality rate was statistically significantly higher (33.3% vs 7.3%, P<0.05), and median EFS (148 d,95%CI 58-238 d vs 372 d,95%CI 264-480 d) (P<0.01) and median OS (179 d 95%CI 6-352 d vs 444 d) (P<0.01) were significantly shorter in NPM1 AML. A total of 126 accompanying gene mutation sites were detected in 87.7% of patients with NPM1AML. The patients with NRAS gene mutation displayed a higher rate of complete remission (100% vs 58%) (P<0.05) and longer median OS (not reached to 320 d, 95%CI 150-490 d) (P<0.05). The 43 fusion genes were examined in 65 out of 73 cases of NPM1AML, and in all the patients the fusion gene test was negative. Multivariate analysis showed that NPM1 VAF>38.4% was an independent prognostic factor for EFS (HR=3.1, 95% CI 1.6-6.4, P<0.01) and OS (HR=3.0, 95% CI 1.4-6.2, P<0.01).
CONCLUSION
The NPM1 gene mutation in AML patients often is accompanied by other gene mutations, while the coexistence of fusion genes is rare; high NPM1 mutant allele burden is an independent prognostic factor for adult AML patients with mutated NPM1.
Alleles
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Humans
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Leukemia, Myeloid, Acute
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genetics
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Mutation
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Nuclear Proteins
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genetics
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Prognosis
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fms-Like Tyrosine Kinase 3
7.Detection of Genetic Mutations in Primary Hypereosinophilia Patients.
Jie ZHOU ; Hao WU ; Bing LI ; Ai-Bin LIANG ; Jian-Fei FU
Journal of Experimental Hematology 2019;27(2):504-508
OBJECTIVE:
To explore the potential pathogenetic mutations of primary hypereosinophilia(HEN)by sequencing FGFR1 FLT3, MPL and JAK2 genes, and to clarify their effect on clinical manifestation and prognosis of HEN patients.
METHODS:
The direct DNA sequencing was employed to detect the gene mutations of FGFR1, FLT3, MPL and JAK2 in HEN patients.
RESULTS:
One deletion mutation (2654_2753del) within tyrosine kinase domain of FLT3 gene was found in a patient suffered from severe symptoms and ended with dismal outcome, which induced a premature stop codon (G885fsX888). For FGFR1, a new variation described as 1014_1019del AACAGT for nucleotide change was found in 19 cases, resulting in T339_V340del at the protein level.
CONCLUSION
The deletion of 6 bases in the FGFR1 gene (1014_1019del AACAGT) is first reported as non-synonymous SNP (nsSNP) site in the patients with primary hypereosinophilia. Deletion mutations in the FLT3 gene may be related with malignant clinical features and poor prognosis.
Base Sequence
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Humans
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Hypereosinophilic Syndrome
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genetics
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Mutation
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Receptors, Thrombopoietin
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Sequence Deletion
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fms-Like Tyrosine Kinase 3
8.Prognostic Significance of FLT3 Internal Tandem Duplication in Acute Myeloid Leukemia with Normal Karyotype.
Ja Young LEE ; Young Don JOO ; Seung Hwan OH ; Jeong Hwan SHIN ; Hye Ran KIM ; Sang Min LEE ; Jeong Nyeo LEE
Korean Journal of Hematology 2009;44(2):74-81
BACKGROUND: Activating mutations of the fms-like tyrosine kinase 3 gene (FLT3) by internal tandem duplication (ITD) in the juxtamembrane region are found in 20~30% of the adults with acute myeloid leukemia (AML). The allelic ratio of FLT3/ITD (ITD-AR), as assessed by Genescan analysis, has been reported to carry prognostic significance in AML patients who have normal karyotype. METHODS: FLT3/ITD was studied by PCR in 113 adults with AML including 55 patients with normal karyotype. Genescan analysis was performed for the PCR products to determine the allelic distribution. The results were correlated with the prognostic factors. RESULTS: FLT3/ITD was found in 23% of the total AML patients and in 32.7% of those patients with normal karyotype. The mutation was related to a high WBC count, a high serum LD level and a low percentage of CD34 positive cells. The ITD-AR ranged from 0.05 to 8.27 (median, 0.61). The patients with a high ITD-AR (> or =0.7) had significantly higher WBC count and LD level than those without FLT3/ITD. On multivariate analysis, a high ITD-AR as well as FLT3/ITD were confirmed to be independent adverse prognostic factors for AML patients with normal karyotype. CONCLUSION: This study demonstrated that a high ITD-AR and FLT3/ITD had major adverse impacts on the prognostic relevance for AML patients with normal karyotype.
Adult
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fms-Like Tyrosine Kinase 3
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Humans
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Karyotype
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Leukemia, Myeloid, Acute
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Multivariate Analysis
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Polymerase Chain Reaction
9.Donor HSCs with a preexisting ASXL1-mutation evoluting FLT3-ITD positive AML-M2 and FLT3-ITD negative AML-M5 in the donor and recipient respectively: one case analysis and literatures review.
Dai KONG ; Kai SUN ; Ming Yue SHI ; Yong Ping SONG ; Mei Ying WEN ; Zhong Wen LIU
Chinese Journal of Hematology 2019;40(4):327-329