Distinctive Spectra of Cytogenetic Alterations in Over Five Hundred East Asian Patients with Acute Leukemia

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4408-4408
Author(s):  
Hyun-Jung Choi ◽  
Hye Ran Kim ◽  
Myung-Geun Shin ◽  
Jong-Hee Shin ◽  
Bock-Hee Park ◽  
...  
Keyword(s):  
Acute Leukemia ◽  
Chromosomal Aberrations ◽  
Molecular Detection ◽  
Conflicts Of Interest ◽  
Detection System ◽  
East Asian ◽  
Normal Karyotype ◽  
Fusion Genes ◽  
Rt Pcr ◽  
Conventional Cytogenetics

Abstract Abstract 4408 Background: Racial difference of spectra of chromosomal aberrations in various hematological malignancies was previously reported between Asian and western countries. Therefore, characterization of the cytogenetic profile is still a matter of interests for identifying cytogenetic risk factors in the current risk-adapted chemotherapy. The aim of this study was to investigate the spectrum of chromosomal aberrations in East Asian (Korean) acute leukemia patients, and to propose a panel of leukemic fusion genes suitable for the development of new molecular detection systems. Patients and Methods: We prospectively analyzed bone marrow samples from 502 patients with acute leukemia referred for screening leukemic fusion genes by simultaneous analysis of conventional cytogenetics, FISH and multiplex RT-PCR system in Chonnam National University Hwasun Hospital (Hwasun, Korea) for last 5 years. Results: A total of 334 patients were diagnosed with AML, 155 with ALL and 13 with mixed phenotype acute leukemia. Twenty types in 28 fusion genes were detected in 42.4% of the total patients by multiplex RT-PCR system and in 33.9% by conventional cytogenetics including FISH. In the group of AML, the common fusion genes were 57 PML/RARa, 38 AML1/MGT8, 12 CBFb/MYH11, 11 MLL1, 5 BCR/ABL1, SET/CAN, PLZF/RARA and TLS/ERG in 2 cases each, and one of each AML1/MDS1, TEL/ABL and TEL/MN1. In the group of ALL, the following fusion genes were detected: 33 BCR/ABL1, 22 TEL/AML1, 10 MLL1, 7 E2A/PBX, 2 SET/CAN and SIL/TAL, one TEL/ABL. In addition, cytogenetically cryptic translocations were detected in 14.8% patients with normal karyotype or numerical aberrations by conventional cytogenetics. Among 288 patients with negative results by multiplex RT-PCR system, one child with early pre-B-ALL harbored MLL1/AF4, and 8 patients had 3 types of clinically significant chromosomal aberrations such as t(3;3)(q21;q26.2), t(8;14)(p24.1;q32) and i(17)(q10), which were all confirmed by conventional cytogenetics. Conclusions: The current study demonstrated the spectrum and frequency of chromosomal aberrations in Korean patients with acute leukemia, which are different from Caucasian data. Also these results may offer important implications for the development of new multiplex molecular detection system. Disclosures: No relevant conflicts of interest to declare.

The Spectra of Chromosomal Aberrations In Acute Leukemia and Implications for the Development of Optimal Profiles of Leukemic Fusion Genes In Multiplex RT-PCR System.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3362-3362
Author(s):  
Hye Ran Kim ◽  
Hyun-Jung Choi ◽  
Hee-Jo Baek ◽  
Hyeoung Joon Kim ◽  
Hoon Kook ◽  
...  
Keyword(s):  
Acute Leukemia ◽  
Chromosomal Aberrations ◽  
Chromosomal Abnormalities ◽  
Conflicts Of Interest ◽  
Fusion Genes ◽  
Rt Pcr ◽  
Negative Results ◽  
National University ◽  
Conventional Cytogenetics ◽  
11Q23 Abnormalities

Abstract Abstract 3362 Background: Cytogenetic aberrations have been identified as important prognostic parameters for acute leukemia, in addition to diagnosis and treatment. The aim of the current study was to investigate the spectrum of chromosomal abnormalities in East Asian (Korean) patients with acute leukemia. Based on the spectrum of genetic aberrations found in our study, we proposed a revised list of chromosomal aberrations for the development of optimal profiles of leukemic fusion genes in multiplex RT-PCR system. Patients and Method: We prospectively analyzed blood or bone marrow samples from 348 patients (260 adults, 88 children) with leukemia from 2006 to 2009 at the Chonnam National University Hwasun Hospital (Hwasun, Korea). Simultaneous analyses of conventional cytogenetics, FISH and a commercially available multiplex RT-PCR system (HemaVision) were performed. Result: Thirty-six types of chromosomal abnormalities were found in the total patient population. Twenty types of chromosomal abnormalities were detected in 42% of all patients by multiplex RT-PCR system and in 33% of all patients by conventional cytogenetics including FISH. Within the group of 213 AML patients, the common cytogenetic abnormalities were t(15;17)(q21;q22) (n=35), t(8;21)(q22;q22) (n=31), 11q23 abnormalities (n=7), inv(16)(p13;q22) (n=3), t(9;9)(q34:q34) (n=2), and the abnormalities t(9;22)(q34;q11), t(9;12)(q34;p13), t(3;21)(q26;q22), t(11;17)(q23;q21), t(12;22)(p13;q11) and t(16;21)(p11.2;q22.3) were each involved in one case. In the group of 104 ALL patients, the following fusion transcripts were detected: 21 t(9;22)(q34;q11), 15 t(12;21)(p13;q22), nine 11q23 abnormalities, four t(1;19)(q23;p13), with t(9;9)(q34:q34), t(9;12)(q34;p13) and del(1p34) each involved in one case. In addition, cryptic translocations were detected in 9% of patients with normal karyotypes or numerical aberrations by conventional cytogenetics. Among 53 patients showing negative results for multiplex RT-PCR, one child patient with early pre-B-ALL harbored MLL1/AF4 fusion genes and 5 patients had 3 types of clinically significant chromosomal aberrations such as t(3;3)(q21;q26.2), t(8;14)(p24.1;q32) and i(17)(q10). Conclusion: The current study demonstrated the spectrum and frequency of chromosomal abnormalities in Korean leukemia patients, which were slightly differed from previous studies including western and some Asian countries. These results may offer important implications in understanding characteristics of molecular pathophysiology, the development of a new RT-PCR system as well as revision of the current commercial available RT-PCR system. Disclosures: No relevant conflicts of interest to declare.

Myelodysplastic Syndromes: A Multidisciplinary Integrated Diagnostic Work-up for Patients' Risk Stratification

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5579-5579
Author(s):  
Elena Ciabatti ◽  
Maria Immacolata Ferreri ◽  
Angelo Valetto ◽  
Alice Guazzelli ◽  
Veronica Bertini ◽  
...  
Keyword(s):  
Myelodysplastic Syndromes ◽  
Mutational Analysis ◽  
Conflicts Of Interest ◽  
Normal Karyotype ◽  
Diagnostic Workup ◽  
Fish Analysis ◽  
Rt Pcr ◽  
Risk Scoring ◽  
Conventional Cytogenetics ◽  
Work Up

Abstract Conventional cytogenetics continues to have a fundamental role in the classification and risk scoring of myelodysplastic syndromes (MDS). Nevertheless, non-informative karyotypes represent up to 20% of cases. Some different molecular methods, not included in the routinary diagnostic workup, such as aCGH or mutational analysis, could be able to detect new abnormalities and improve the subtyping of MDS. The aim of this study was to propose a new diagnostic workup to determine the eventual adjunctive value offered by FISH, aCGH, and somatic mutation assays in respect of the the conventional cytogenetics only. In this study, we analyzed 50 patients: 29% female and 71% male, median age 71 (range 30-88 years), 66% at low/int1 IPSS risk, 54% at very-low/low R-IPSS risk, 33% with RCMD, 15% with RA, 14% with RARS, 14% with RAEB, 8% 5q-, and 16% with MMCL. We assessed these new MDS cases by different techniques: i) conventional cytogenetics; ii) FISH for chromosome 5, 7, PDGFRa, and PDGFRb; iii) aCGH, and iiii) specific RT-PCR for ASXL1, EZH2, TP53, and TET2 mutations. Conventional cytogenetics showed 42% of patients with at least one chromosomal aberration, including +8, del(11), del(7), del(5), -Y, +6, del(13), +14, del(20), and complex karyotypes (6% of cases). After FISH analysis, we were able to correctly classify as affected by the 5q- syndrome 2 cases who then received lenalidomide. The aGCH allowed to detect quantitative chromosomal aberrations in 44% of cases (del(13), -7, del(12), del(16), del(17), del(11), del(8), dupl(14), 5q-), including 10 cases (20%) showing a normal karyotype. After the RT-PCR, 32% of patients resulted mutated, with highest frequency for TP53 (22%). Four of these TP53-mutated patients showed normal karyotype, and resulted unmutated also by FISH and aCGH; in a case TP53 mutated we added treatment with steroid. Other 3 patients TP53-mutated did not respond to azacitidine Four low-risk patients (8%) showed ASXL1 gene mutation, three of them not earlier detected by cytogenetics or aCGH. One of these patients died after progression into acute leukemia. The identification of TP53 or ASXL1 mutations after RT-PCR and of dupl(14) by the aCGH prompted us to strictly follow patients at high risk of transformation. Only one case showed TET2 mutation; although TET2 mutations have been related to a better survival in patients receiving 5-azacitidine, this patient resulted not-responsive after 9 cycles. In conclusion, these results sustain the necessity of an integrated work-up for the diagnosis and the correct risk scoring of MDS patients. Disclosures No relevant conflicts of interest to declare.

Analysis of Genetic Alterations In 843 Korean Acute Leukemia Patients by Multiplex RT-PCR Assay (HemaVision) and Karyotyping

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4845-4845
Author(s):  
Jihyang Lim ◽  
Myungshin Kim ◽  
Yonggoo Kim ◽  
Kyungja Han ◽  
Hee-Je Kim ◽  
...  
Keyword(s):  
Acute Leukemia ◽  
Conflicts Of Interest ◽  
Simultaneous Detection ◽  
Normal Karyotype ◽  
Genetic Alterations ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Abnormal Karyotype ◽  
Acute Leukemias ◽  
Positive Results

Abstract Abstract 4845 Background: The importance of molecular tests has been increased for diagnosis, classification and MRD monitoring in acute leukemia. Recently, multiplex RT-PCR assays (HemaVision) for simultaneous detection of 28 leukemia-associated translocations was introduced and has been used for molecular screening of leukemic patients. We analyzed the results of multiplex RT-PCR assay (HemaVision) and karyotyping in Korean acute leukemia patients. Methods: From April 2007 to March 2010, multiplex RT-PCR assay (HemaVision, DNA Technology A/S, Denmark) was performed in 843 Korean acute leukemia cases (AML 566 vs. ALL 277 and adults 596 vs. pediatrics 237) for screening leukemia-associated translocations. All results were compared with chromosome analysis results by conventional karyotyping. Results: Three hundreds fifty-one cases (41.6%) were positive in multiplex RT-PCR. In AML cases, 221 cases (39.0%) revealed positive results and RUNX1-RUNX1T1 85 cases (38.5%), PML-RARA 74 cases (33.5%), CBFB-MYH11 25 cases (11.3%) and MLL-related 19 cases (8.6%) were frequently detected. In contrast, 130 cases (46.9%) revealed positive results in 277 ALL cases and BCR-ABL1 68 cases (52.3%), ETV6-RUNX1 28 cases (21.5%), TCF3-PBX1 15 cases (11.5%), MLL-related 10 cases (7.7%) were frequently detected. In comparison to karyotyping, 337 cases (96.0%) revealed abnormal karyotype and 14 cases (4.0%) revealed normal karyotype in 351 cases that revealed positive results by multiplex RT-PCR assay. In 492 cases that revealed negative results by multiplex RT-PCR assay, 278 cases (56.5%) revealed abnormal karyotype and 214 cases (43.5%) revealed normal karyotype. Overall 629 cases (74.6%) were detected genetic alterations by multiplex RT-PCR assay and/or karyotyping. Conclusion: Most acute leukemias may have a genetic alteration in their leukemogenesis. Both multiplex RT-PCR assay and karyotyping are essential for screening leukemia-associated genetic alterations. Disclosures: No relevant conflicts of interest to declare.

Application of Multiplex RT-PCR for Identification of Leukemia Associated Gene Abnormalities.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4431-4431
Author(s):  
Hyun Woo Lee ◽  
Youn Mu Jung ◽  
Jun Ho Jang ◽  
Jun Seong Park ◽  
Sung Ran Cho ◽  
...  
Keyword(s):  
Acute Leukemia ◽  
Normal Karyotype ◽  
Concordance Rate ◽  
Molecular Probe ◽  
Rt Pcr ◽  
Prognostic Gene ◽  
Number Of Patients ◽  
Genetic Abnormalities ◽  
Pcr Method ◽  
Cytogenetic Technique

Abstract The best prognostic predictor for acute leukemia is known to be the finding of genetic abnormalities of leukemic cells. Methods for detecting the genetic abnormalities include chromosomal studies for karyotyping, FISH(Fluorescence in situ hybridization) and RT-PCR. However, each methods have limitations i.e. low sensitivity in karyotyping, uncertainty of molecular probe to be used in FISH or RT PCR methods. Multiplex RT-PCR (MRT-PCR) allows simultaneous detection of 28 fusion genes, more than 80 breakpoints and splice variants associated with leukemia. Therefore, this method can be used for detection of molecular abnormality in fresh unknown leukemic cases, as well as for molecular remission in follow-up cases. The aim was to demonstrate whether MRT-PCR system might be successfully used to screen a large number of patients with acute leukemia and compare the result with that of chromosome studies. Frozen bone marrow cells from 78 patients, who were diagnosed with acute leukemia at Ajou university hospital between September 1994 and February 2004, were used for MRT-PCR. In all samples with a known conventional cytogenetic results, we performed MRT-PCR and to compared with conventional cytogenetic study regarding the concordance rate and analyzed discordant cases regarding their types. 78 samples(40 male and 38 female patients) were analyzed, and there were 59 AML patients and 19 ALL patients. We successfully obtained the mRNA from all frozen samples. In 21 cases with gene abnormalities by chromosome studies most of them (15/21) showed the same abnormalities with MRT-PCR. In 57 patients with normal karyotype by cytogenetic technique, we identified 18 translocations of clinical significance by MRT-PCR method. In 18 discordant cases, there were 4 cases with t(15;17), 4 cases with t(8;21), 3 cases with t(9;22), 2 cases with t(11;19) and 4 others [t(9;11),t(9;9),t(3;11),inv(16)]. Conventional cytogenetics detected 10 cases of good prognostic gene abnormalities [t(15;17),t(8;21),inv(16)], but MRT-PCR method detected 10 additional cases of good prognostic gene abnormalities which might change the treatment paln. The twenty good prognostic cases with MRT-PCR showed better overall survival than others. (median f/u = 10.6 month, p=0.0443) There were 69% concordance rate between cytogenetic technique and MRT-PCR. Furthermore clinically significant translocations were detected by MRT-PCR in 18 of 57 normal karyotype patients, indicating improved sensitivity and prognostic value with MRT-PCR. Further investigations are needed to ascertain the usefulness of MRT-PCR for the screening tool of leukemic gene abnormalities.

Highly Multiplexed Targeted Sequencing of Recurrent Fusion Genes in Acute Leukemia

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 2335-2335
Author(s):  
Philippe Ruminy ◽  
Nimrod Buchbinder ◽  
Thomas Larson ◽  
Vinciane Marchand ◽  
Bérangère Joly ◽  
...  
Keyword(s):  
Acute Leukemia ◽  
Low Cost ◽  
Clinical Information ◽  
Daily Practice ◽  
Leukemic Cells ◽  
Correct Identification ◽  
Sequencing Analysis ◽  
Fusion Genes ◽  
Rt Pcr ◽  
Childhood All

Abstract In acute leukemia, recurrent chromosomal translocations which result in the fusion of two genes are frequent. Some of these markers have a well established prognostic and therapeutic impact and are routinely screened at diagnosis by cytogenetics and RT-PCR. Yet, due to the limitations of these methods, only a few among the dozens of known rearrangements are systematically tested. Many abnormalities which could provide important clinical information thus remain ignored, mainly due to the impossibility of performing a cost effective multi-targeted screening. To overcome this obstacle, we have developed a simple and parsimonious assay which allows a reliable detection of dozens of fusion genes in only a few hours. Our assay was designed to screen simultaneously more than 50 translocations involving 70 genes, recurrent in acute myeloid (AML), acute lymphoid (ALL) and chronic myeloid (CML) leukemia. cDNA samples obtained from leukemic cells are first incubated with a mix of oligonucleotides probes which are complementary to the ends of the exons, at the abnormal junctions on the fusion mRNAs (Figure). For most genes, different probes were designed to detect different transcripts resulting from alternative genomic breakpoints (for example on exons 1, 13, 14 and 19 for BCR, and on exons 2 and 3 for ABL). The mix we created thus regroups more than 150 probes and targets more than 400 different fusion transcripts. All left probes have a common tail (T1) at their 5' ends, all right probes a common tail (T2) at their 3' ends. Additional probes were also included to detect the most frequent mutations of NPM1 (A, B, D). If one translocation is present in the sample, two probes hybridize side to side on the fusion cDNA. A DNA ligase is next used to create a covalent link between these probes, allowing their amplification by PCR with the T1 and T2 primers. If a PCR product is amplified, the two partners are identified by sequencing analysis. To test this method, we applied it to a retrospective series of 430 patients, (252 AML and 178 childhood ALL). In B-ALL (147 cases), all 33 ETV6-RUNX1, 6 BCR-ABL and 5 TCF3-PBX1 as well as 6 MLL rearrangements (3 AF4; 1 ENL, 1 AF9 and 1 AFF4) identified at diagnosis by conventional methods were detected, as well as 5 previously unnoticed P2RY8-CRLF2 junctions. We only failed to detect one MLL-AF4 fusion detected by cytogenetics but not by RT-PCR, probably due to the poor quality of the RNA. Further analysis of 10 MLL-AF4 positive adult ALL patients allowed the correct identification of this fusion in all cases. In T-ALL (31 cases), we detected 6 known (4 SIL-TAL, 2 CALM-AF10) and 5 previously undetected fusions (2 NUP214-ABL, 1 MLL-ENL, 1 ETV6-ABL, and a new PLZF-ABL junction). In AML, we detected 86 fusions: 23 PML-RARA, 2 PLZF-RARA, 18 CBFB-MYH11, 12 RUNX1-RUNX1T1, 4 NUP98-NSD1, 2 BCR-ABL, 1 DEK-NUP214, 1 CALM-AF10, 1 MOZ-CBP, 22 MLL rearrangements (13 PTD, 3 AF9, 2 AF6, 1 AF10, 1 ENL, 1 AF1Q and 1 MAPRE) and 44 NPM1 mutations. Two PML-RARA cases, one with a BCR2 breakpoint (not included in our design), one weakly positive by RT-PCR but negative by cytogenetics, two t(11;17) validated by FISH but negative by RT-PCR, and one NPM1J mutation (not included in our assay) were not detected. Importantly, 20 translocations in this series, including 14 MLL fusions and a cytogenetically cryptic t(8;21) had not been identified at diagnosis. Furthermore, all these new abnormalities (in AML and ALL) could be confirmed by conventional RT-PCR and sequencing analysis, demonstrating the specificity of the method. In the whole cohort of 430 patients, the three methods thus detected 157 fusions. 85 (54.1%) and 112 (71.3%) had been detected at diagnosis by cytogenetics and RT-PCR respectively, and 152 (96.8%) by our assay. Two were revealed only by cytogenetics, one only by RT-PCR and 30 only by our method. In conclusion, we have developed a simple multiplexed assay which can reveal a very large number of recurrent gene fusions in leukemia. Its short turn-around time (we repeatedly tested up to 40 patients in parallel and the results can be obtained in less than one day) and low cost (only one PCR module, one pyrosequencer and basic molecular biology reagents are needed) make it particularly suitable for a daily practice. Its capacity to detect many abnormalities which are almost never tested in daily practice could provide many important diagnosis and prognosis information, and enable the stratification of patients in prospective clinical trials. Figure 1 Figure 1. Disclosures Ruminy: Centre Henri Becquerel: Patents & Royalties. Marchand:Centre Henri Becquerel: Patents & Royalties.

Multiplex Reverse Transcription-Polymerase Chain Reaction for Simultaneous Screening of 29 Translocations and Chromosomal Aberrations in Acute Leukemia

Blood ◽  
1998 ◽  
Vol 92 (2) ◽  
pp. 574-588 ◽  
Author(s):  
Niels Pallisgaard ◽  
Peter Hokland ◽  
Dorthe C. Riishøj ◽  
Bent Pedersen ◽  
Poul Jørgensen
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Leukemia ◽  
Chromosomal Aberrations ◽  
Reverse Transcription ◽  
Cytogenetic Analysis ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Lymphoid Leukemia ◽  
Polymerase Chain ◽  
Set Up

Abstract We have developed a multiplex reverse transcription-polymerase chain reaction (RT-PCR) reaction, which enables us to detect 29 translocations/chromosomal aberrations in patients with acute lymphoid leukemia (ALL) and acute myeloid leukemia (AML). Through the construction and optimization of specific primers for each translocation, we have been able to reduce the set-up to 8 parallel multiplex PCR reactions, thus greatly decreasing the amount of work and reagents. We show the value of our set-up in a retrospective analysis on cryopreserved material from 102 AML and 62 ALL patients. The multiplex RT-PCR detected a hybrid mRNA resulting from a structural chromosomal aberration in 45 of 102 (44%) of the AML and in 28 of 62 (45%) of the pediatric ALL cases. Importantly, in 33% of AML and in 47% of the ALL cases with cytogenetic data, submicroscopic chromosomal aberrations or masked translocations were shown that were not detected in the cytogenetic analysis either for structural reasons or because of an insufficient number of metaphases obtained. This multiplex RT-PCR system, which can handle up to 10 patients with a response time of 2 working days, is thus an important tool that complements cytogenetic analysis in the up-front screening of acute leukemia patients and should provide a rapid and efficient characterization of leukemia cells, even in situations with sparse patient material.

Complementary Diagnostic and Prognostic Value of Parallel Karyotyping and Molecular Diagnostics in Patients with Acute Leukemias.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4527-4527
Author(s):  
Anna Ejduk ◽  
Miroslaw Majewski ◽  
Iwona Solarska ◽  
Hanna Makuch-Lasica ◽  
Iwona Kania ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Molecular Diagnostics ◽  
Chromosomal Aberrations ◽  
Risk Group ◽  
Treatment Options ◽  
Molecular Methods ◽  
Fusion Genes ◽  
Rt Pcr ◽  
Acute Leukemias ◽  
Molecular Tests

Abstract Chromosomal aberrations analyzed by cytogenetic and molecular methods are major prognostic factors determining treatment options in patients with acute leukemias. The aim of this study was to compare cytogenetic and molecular methods as diagnostic and prognostic tools in patients with acute leukemias. Sixty one previously untreated patients with acute leukemias (AML - 43, ALL - 18) were studied for the presence of chromosomal translocations and corresponding fusion genes [AML1-ETO t(8;21), CBFB-MYH11 inv(16), PML-RARA t(15;17), BCR-ABL p190, p210 t(9;22), MLL-AF4 t(4;11), TEL-AML1 t(12;21), E2A-PBX1 t(1;19), SIL-TAL1 del(1)] using both molecular (RT-PCR, nested PCR) and cytogenetic (GTG) methods. Molecular diagnostics was performed from RNA isolated from bone marrow samples according to BIOMED-1 protocol. Cytogenetic studies were carried out with classical GTG and FISH methods. G-banded mitoses of bone marrow specimen were analysed according to ISCN. Chromosomal aberrations were found in 32.8% patients using GTG method while the parallel molecular tests revealed related fusion genes in 50.8% patients. In 8.5% patients cytogenetic analysis was not performed because of lack of metaphases in cultured cells. All cytogenetic aberrations found in GTG were also confirmed in RT-PCR. Stratification into cytogenetic risk groups was performed after applying combined analysis of karyotyping and molecular tests. Low cytogenetic risk group consisted of 32.8% patients including 11.5% diagnosed with GTG and additionally 21.3% patients after applying molecular tests. The intermediate and high cytogenetic risk group consisted of 32.8% and 34.4% respectively using combined cytogenetic and molecular diagnostics. In low cytogenetic risk group, 85% of patients achieved complete remission (CR), early deaths were found in 15% and none of the patients presented primary chemotherapy resistance. In intermediate risk group CR were obtained in 80%, chemoresistance in 10% and early deaths were observed in 10% of patients. In high cytogenetic risk group, CR were achieved in only 23.8% and chemoresistance occured in 76.2% of the patients. In conclusion we suggest that molecular and cytogenetic tests are complementary methods and should be used in parallel in the initial diagnosis of patients with acute leukemias. This seems to be critical for obtaining the accurate diagnosis, cytogenetic risk assessment and choosing an optimal treatment options.

scholarly journals Frequency of p190 and p210 BCR-ABL Fusions Genes in Acute Lymphoblastic Leukemia in a Long Group of Adults and Childhood

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 5273-5273 ◽  
Author(s):  
Rosa Maria Arana-Trejo ◽  
Gregorio Ignacio ◽  
Raquel Amador-Sánchez ◽  
Jorge Cruz-Rico ◽  
Maria-Paula Hernández ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
De Novo ◽  
Conflicts Of Interest ◽  
Lymphoblastic Leukemia ◽  
Fusion Transcript ◽  
Age Group ◽  
Fusion Genes ◽  
Rt Pcr ◽  
Pcr Multiplex ◽  
The Impact

Abstract The Ph chromosome is a translocation (9;22)(q34;q11), that results in the constitutive activation of the BCR/ABL tyrosine kinase. The incidence of BCR/ABL in Acute Lymphoblastic Leukemia (ALL) increases with age, from less than 5% in younger children to 20-30% in older adults, with a peak incidence in patients aged 35-50 years. BCR/ABL1 has diverse breakpoints, in adult patients with Ph+ ALL the p190BCR/ABL transcript e1a2/e3a2 may be documented in 50-70%; p210BCR/ABL b2a2/b3a2 in 15-30% of patients and <1% having both breakpoint. Childhood patients with Ph+ ALL fusion genes present p190BCR/ABL transcipt e1a2/e3a2 in 90% and the remaining present other fusion transcrit or co-expression of both p190 and p210 BCR-ABL. OBJETIVE. The aim of this study was identify the occurrence of fusion genes to p190 and p210 BCR-ABL rearrangements in adult and childhood patients with ALL. METHODS. We include between 2008-2015 870 patients with ALL de novo from seven different hospitals from México, the 45% (394) were childood and the rest 55% (476) were adults. All patients were studied to RT-PCR multiplex and nested in RNA for fusion transcripts 190 and p210 BCR-ABL, at diagnosis, according to the international BIOMED-1 protocol. RESULTS. From 870 patients with ALL, the most frequent subtype FAB were L2 (87%) and second L1 (13%). The immunophenotype by B-ALL was to 80%, bilineal in 5% and the rest have not data. The overall incidence to BCR-ABL in both children and adults with ALL were to 17% [147/870]. The analysis by age group were; in 476 adults with ALL, their average age was 37 years old (range 17-84 years) and their incidence of BCR-ABL positive was 20% (95/476 cases). The distributions by type of fusion transcript were 83% p190 and 17% p210; we did not observe co-expression of transcripts to BCR-ABL. In children patients the average age was 9 years old (range 0.1-16 years), the incidence of BCR-ABL was 13.2% (52/394 cases). The distributions by type of fusion transcript to BCR-ABL were p190 78.8%; p210 13.4% and their co-expression by both isoforms 8%. CONCLUSION. The 20% frequency for BCR-ABL1 in adults with ALL is concordant with others reports published, with values from 17% to 37% with predominancy of p190 (83%). In our pediatric patients group with ALL, document a frequency of 13.2% by BCR-ABL1 positive; it is higher than other populations reporting 5-10%. The distributions of fusion transcript p190 and p210 coincides with previous prevalence estimates in other countries where p190 transcript was the most frequent. But the coexpression of both isoforms [p190/p210] were 8% it has not been reported in this age group with ALL. In conclusion, we recommend to identify the BCR-ABL transcript type in every patients with ALL at diagnosis, using a RT-PCR verified method for P190/p210 and followed the patient by mesure the impact clinical and will be adjust the treatment like o plus the cytogenetic studies. Disclosures No relevant conflicts of interest to declare.

scholarly journals FLT3, NPM1 and CEBPA Mutations in 195 Children with Acute Myeloid Leukemia in Argentina

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5318-5318
Author(s):  
Cristina N. Alonso ◽  
Patricia L. Rubio ◽  
Adriana Medina ◽  
Silvia Eandi Eberle ◽  
Andrea Bernasconi ◽  
...  
Keyword(s):  
Conflicts Of Interest ◽  
Prognostic Significance ◽  
Normal Karyotype ◽  
Prognostic Impact ◽  
Sequencing Analysis ◽  
Rt Pcr ◽  
Genetic Abnormalities ◽  
Childhood Aml ◽  
The Mean ◽  
Cebpa Mutations

Abstract Background: Mutations of FLT3, NPM1 and CEBPA are found in 25 to 35% of adult-AML. These mutations correlate with outcome, especially in AML with normal karyotype. There are few reports concerning the incidence and prognostic significance of these mutations in childhood-AML and there is no data from Argentina. Objectives: To describe the prevalence of FLT3, NPM1 and CEBPA mutations and to analyze the prognostic impact in the outcome in our setting. Methods: Samples from 195 children treated with AML protocols were retrospectively analyzed. The mean age at diagnosis was 6.8 [0.0-17.9] years, including 65 patients younger than 2 years of age. FAB subtypes were M2: 18%, M3: 15%, M4: 12%, M5: 34%, M6: 3%, M7: 10%, while 16 cases (8%) disclosed an ambiguous lineage immunophenotype. Genetic abnormalities of AML cases were characterized by cytogenetic analysis (97%) and/or RT-PCR for AML1-ETO, CBFB-MYH11, PML-RARA, MLL-AF4, MLL-AF9, MLL-ENL and MLL-AF10 fusion transcripts (95%). The distribution of the genetic abnormalities was: AML1-ETO: 11%, PML-RARA: 15%, CBFB-MYH11: 6%, MLL/11q23: 23%, other abnormalities: 25% and normal karyotype: 16%. Detection of NPM1 and CEBPA mutations was performed by Gene-scanning; FLT3-ITD and FLT3-TKD were studied by RT-PCR and RFLP respectively. Positive cases were further characterized by sequencing analysis. Results: The prevalences of the studied mutations were: FLT3-ITD: 10.3%, FLT3-TKD: 8.2%, NPM1mut: 4.6% and CEBPAmut: 2.1%. Within the group of AML with normal karyotype the incidences were: FLT3-ITD: 12.5%, FLT3-TKD: 6.3%, NPM1mut: 25.0% and CEBPAmut: 12.5%. The mean age for each subgroup was: FLT3-ITD: 14 years, FLT3-TKD: 9 years, NPM1mut: 12 years and CEBPAmut: 12 years. Simultaneous presence of FLT3-ITD and NPM1 mutations was detected in 2 cases while 1 patient disclosed both FLT3-TKD and CEBPAmut. FLT3-ITD and FLT3-TKD showed significant association with the presence of PML-RARA (p<0.00001 and p=0.055 respectively). Eight out of nine patients with NPM1mut and 4/4 patients with CEBPAmut were AML with normal karyotype. The FAB subtypes more frequently observed for each subgroup were: FLT3-ITDmut: M3 (n:10/20; p<0.00001), FLT3-TKDmut: M5 (n:8/16; p=n.s.), NPM1mut: M2 (n:4/9; p=0.062) and CEBPAmut: M2 (n:3/4; p=0.019). The mean ages of patients with FLT3-ITDmut, NPM1mut and CEBPAmut were significantly higher (p<0.00001, p=0.006 and p=0.033, respectively). FLT3-TKD was the only mutation detected in 5/45 (11%) of patients younger than 1 year of age. The five-years leukemia-free survival probabilities (pLFS) and standard error (SE) were: Total AML: 49 (4)%, FLT3-ITDmut:68 (12)%, FLT3-TKDmut:46 (17)%, NPM1mut: 75 (15)%, CEBPAmut: 100 (0)% and NPM1mut/CEBPAmut/FLT3-ITDneg: 83 (15)% (p<0.00001). The pLFS (SE) of patients with normal karyotype and FLT3-ITDneg and NPM1mut or CEBPAmut was 88 (12)% (p=0.066). Conclusions: This is the first report of the frequencies of FLT3, NPM1 and CEBPA mutations in childhood AML in our country. The incidences of NPM1mut and CEBPAmut were significantly higher in AML with normal karyotype. Our data confirm the favorable prognosis of AML with NPM1mut/FLT3-ITDneg and CEBPAmut/FLT3-ITDneg genotypes, especially in cases with normal karyotype. The present results support the notion that this group should be considered as a new AML subset with better outcome. This group of AML patients with better outcome could be included in the standard risk group, thus avoiding intensive treatments and related toxicity. Disclosures No relevant conflicts of interest to declare.

Comparative analysis of chromosomal aberrations in children with acute leukemia

2020 ◽  
Vol 5-6 (215-216) ◽  
pp. 7-14
Author(s):  
Zhansaya Nessipbayeva ◽  
◽  
Minira Bulegenova ◽  
Meruert Karazhanova ◽  
Dina Nurpisova ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Acute Leukemia ◽  
Chromosomal Aberrations ◽  
Cytogenetic Study ◽  
Normal Karyotype ◽  
Blast Cell ◽  
Bone Marrow Blast ◽  
Scientific Center ◽  
Blast Cells ◽  
Medical Histories

Leukemia is a hematopoetic tissue tumor with a primary lesion of the bone marrow, where the morphological substrate is the blast cell. Chromosomal and molecular genetic aberrations play a major role in the acute leukemia pathogenesis, determing the morphological, immunological and clinical features of the disease. Our study was aimed to to analyze retrospectively the structure and frequency of chromosomal aberrations in children with initially diagnosed acute leukemia. Material and methods. Medical histories retrospective analysis of children charged to oncohematology department of the «Scientific Center of Pediatrics and Pediatric Surgery» in Almaty for the period 2015 - 2017 was carried out. 310 histories with primary diagnosed acute leukemia were studied. Results and discussion. Among 310 patients different chromosome aberrations were isolated in 158 patients (51%) during cytogenetic and molecular cytogenetic (in situ hybridization) studies of bone marrow blast cells. A normal karyotype was observed in 102 patients (33%). Conclusion. The lymphoblastic variant of acute leukemia was determined in 75.5%, that indicates its leading role in AL structure among the children of different ages. AML was determined in 22.6% of all OL cases. The most frequent chromosomal rearrangement in ALL patients was blast cell chromosome hyperdiploidy (10,6%) and t(12;21)(p13;q22)/ETV6-RUNX1,which was detected in 37 (16%) patients. The most frequent AML abberation was t (8;21) (q22;q22)/RUNX1-RUNX1T1, identified in 15 (21.4%) patients. Keywords: acute leukemia, bone marrow, blast cells, karyotype, chromosomal aberrations, cytogenetic study.

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