Sequential Interphase FISH Analysis of m-BCR/ABL Chimeric Gene-Positive Cells in Ph-Positive Acute Myeloid Leukemia

Fusion between the NUP98 and NSD3genes in a patient with acute myeloid leukemia associated with t(8;11)(p11.2;p15), is reported for the first time. The t(8;11)(p11.2;p15) was identified by classical cytogenetics. Fluorescence in situ hybridization (FISH) analysis revealed a split signal with a mix of BAC 118H17 and 290A12, indicating the translocation disrupted NUP98. FISH restriction at 8p11-12 showed a split of BAC 350N15. Molecular investigations into candidate genes in this BAC showed the NUP98 fusion partner at 8p11.2 was the NSD3 gene. To date the NSD3 gene has never been implicated in hematologic malignancies.

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Acute Differentiated Dendritic Cell Leukemia: A New Form of Pediatric Acute Myeloid Leukemia (AML) with MLL Translocations.

Blood ◽

10.1182/blood.v106.11.3263.3263 ◽

2005 ◽

Vol 106 (11) ◽

pp. 3263-3263

Author(s):

Luca Lo Nigro ◽

Laura Sainati ◽

Anna Leszl ◽

Elena Mirabile ◽

Monica Spinelli ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Dendritic Cells ◽

Dendritic Cell ◽

Myeloid Leukemia ◽

Fusion Transcript ◽

Fish Analysis ◽

Rt Pcr ◽

Pediatric Acute Myeloid Leukemia ◽

Pediatric Aml ◽

Acute Myeloid

Abstract Background: Myelomonocytic precursors from acute or chronic leukemias can differentiate to dendritic cells in vitro, but leukemias with a dendritic cell immunophenotype are rare, have been reported mainly in adults, and their molecular pathogenesis is unknown. Dendritic cells are classified as Langherans, myeloid and lymphoid/plasmacytoid cells, but leukemias arising from dendritic cells are unclassified in the FAB system. We identified a new entity of pediatric acute myeloid leukemia (AML) presenting with morphologic and immunophenotypic features of mature dendritic cells, which is characterized by MLL gene translocation. Methods and Results: Standard methods were used to characterize the morphology, immunophenotype, karyotype and MLL translocations in 3 cases of pediatric AML. The patients included two boys and one girl diagnosed with AML between 1–6 years old. Their clinical histories and findings included fever, pallor, abdominal and joint pain, adenopathy, hepatosplenomegaly, normal WBC counts but anemia and thrombocytopenia. and no evidence of CNS disease. The bone marrow aspirates were hypocellular and replaced completely by large blasts with irregular nuclei, slightly basophilic cytoplasm, and prominent cytoplasmic projections. There were no cytoplasmatic granules or phagocytosis. Myeloperoxidase and alpha napthyl esterase reactions were negative, excluding FAB M5 AML, and the morphology was not consistent with any standard FAB morphologic diagnosis. The leukemic blasts in all three cases were CD83+, CD86+, CD116+, consistent with differentiated myeloid dendritic cells, and did not express CD34, CD56 or CD117. MLL translocations were identified in all 3 cases. In the first case FISH analysis showed t(10;11)(p12;q23) and RT-PCR identified and a ‘5-MLL-AF10-3’ fusion transcript. In the second case FISH analysis showed t(9;11)(p22;q23) and RT-PCR identified and a ‘5-MLL-AF9-3’ fusion transcript. In the remaining case, the MLL gene rearrangement was identified by Southern blot analysis and RT-PCR showed an MLL-AF9 fusion transcript. The fusion transcripts in all 3 cases were in-frame. Remission induction was achieved with intensive chemotherapy, and all three patients have remained in durable remission for 30–60 months after hematopoietic stem cell transplantation. Conclusions. We have characterized a new pediatric AML entity with features of mature dendritic cells, MLL translocation and an apparently favorable prognosis. The in-frame MLL fusion transcripts suggest that chimeric MLL oncoproteins underlie its pathogenesis. The partner genes in all 3 cases were known partner genes of MLL that encode transcription factors. This study increases the spectrum of leukemias with MLL translocations. Comprehensive morphological, immunophenotypic, cytogenetic and molecular analyses are critical for this diagnosis, and will reveal its frequency and spectrum as additional cases are uncovered.

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Cytogenetically Unrelated Clones in Acute Myeloid Leukemia Showing Different Responses to Chemotherapy

Case Reports in Hematology ◽

10.1155/2016/2373902 ◽

2016 ◽

Vol 2016 ◽

pp. 1-5

Author(s):

Kohei Kasahara ◽

Masahiro Onozawa ◽

Naohiro Miyashita ◽

Emi Yokohata ◽

Miho Yoshida ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Induction Therapy ◽

Myeloid Leukemia ◽

De Novo ◽

Disease Status ◽

Fish Analysis ◽

Monosomy 7 ◽

Appropriate Treatment ◽

Correct Understanding ◽

Acute Myeloid

We report a case of acute myeloid leukemia (AML) with two cytogenetically unrelated clones. The patient was a 45-year-old male who was diagnosed with acute monoblastic leukemia (AMoL). Initial G-band analysis showed 51,XY,+6,+8,inv(9)(p12q13)c,+11,+13,+19[12]/52,idem,+Y[8], but G-band analysis after induction therapy showed 45,XY,-7,inv(9)(p12q13)c[19]/46,XY,inv(9)(p12q13)c[1]. Retrospective FISH analysis revealed a cryptic monosomy 7 clone in the initial AML sample. The clone with multiple trisomies was eliminated after induction therapy and never recurred, but a clone with monosomy 7 was still detected in myelodysplastic marrow with a normal blast percentage. Both clones were successfully eliminated after related peripheral blood stem cell transplantation, but the patient died of relapsed AML with monosomy 7. We concluded that one clone was de novo AMoL with chromosome 6, 8, 11, 13, and 19 trisomy and that the other was acute myeloid leukemia with myelodysplasia-related changes　(AML-MRC) with chromosome 7 monosomy showing different responses to chemotherapy. Simultaneous onset of cytogenetically unrelated hematological malignancies that each have a different disease status is a rare phenomenon but is important to diagnose for a correct understanding of the disease status and for establishing an appropriate treatment strategy.

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RT-PCR and FISH analysis of acute myeloid leukemia with t(8;16)(p11;p13) and chimeric MOZ and CBP transcripts: breakpoint cluster region and clinical implications

Leukemia ◽

10.1038/sj.leu.2403353 ◽

2004 ◽

Vol 18 (6) ◽

pp. 1115-1121 ◽

Cited By ~ 36

Author(s):

H H Schmidt ◽

S Strehl ◽

D Thaler ◽

D Strunk ◽

H Sill ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Clinical Implications ◽

Fish Analysis ◽

Rt Pcr ◽

Breakpoint Cluster Region ◽

Cluster Region ◽

Acute Myeloid

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Coexpression of NUP98/TOP1 and TOP1/NUP98 in de novo Acute Myeloid Leukemia with t(11;20)(p15;q12) and t(2;5)(q33;q31)

Cytogenetic and Genome Research ◽

10.1159/000458168 ◽

2016 ◽

Vol 150 (3-4) ◽

pp. 287-292

Author(s):

Katsuya Yamamoto ◽

Yosuke Minami ◽

Kimikazu Yakushijin ◽

Yu Mizutani ◽

Yumiko Inui ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

De Novo ◽

Fusion Gene ◽

Fusion Transcript ◽

Fish Analysis ◽

Balanced Translocation ◽

Cytogenetic Aberration ◽

De Novo Aml ◽

Acute Myeloid

The t(11;20)(p15;q11∼12) translocation is a very rare but recurrent cytogenetic aberration that occurs in myelodysplastic syndrome/acute myeloid leukemia (MDS/AML). This translocation was shown to form a fusion gene between NUP98 at 11p15 and TOP1 at 20q12. Here, we describe a new case of de novo AML M2 with t(11;20) which was associated with another balanced translocation. An 81-year-old man was admitted to undergo salvage therapy for relapsed AML. G-banding and spectral karyotyping showed 46,XY,t(2;5)(q33;q31),t(11;20)(p15;q12)[20]. Expression of the NUP98/TOP1 fusion transcript was confirmed: NUP98 exon 13 was in-frame fused with TOP1 exon 8. The reciprocal TOP1/NUP98 fusion transcript was also detected: TOP1 exon 7 was fused with NUP98 exon 14. After achieving hematological complete remission, the karyotype converted to 46,XY,t(2;5)(q33;q31)[19]/46,sl,t(11;20)(p15;q12)[1]. FISH analysis demonstrated that the 5q31 breakpoint of t(2;5) was centromeric to EGR1. In all 10 cases described in the literature, the NUP98 exon 13/TOP1 exon 8 fusion transcript was expressed, indicating that it may be responsible for the pathogenesis of MDS/AML with t(11;20). On the other hand, the TOP1/NUP98 transcript was coexpressed in 4 cases of de novo AML, but not in 3 cases of therapy-related MDS. Thus, this reciprocal fusion may be associated with progression to AML.

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The importance of cooperation between cytogenetic and molecular laboratory in the genetic testing of hemato-oncological diseases on the example of a pediatric patient with acute myeloid leukemia (AML)

Diagnostyka Laboratoryjna ◽

10.5604/01.3001.0013.7751 ◽

2019 ◽

Vol 54 (4) ◽

pp. 225-232

Author(s):

Teofila Książek ◽

Katarzyna Szewczyk ◽

Agnieszka Grabowska ◽

Mirosław Bik-Multanowski ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Pediatric Patient ◽

Myeloid Leukemia ◽

Correct Diagnosis ◽

Optimal Treatment ◽

Leukemic Cells ◽

Molecular Karyotyping ◽

Fish Analysis ◽

Genetic Changes ◽

Acute Myeloid

The genetic profiling of leukemic cells is currently the standard in routine diagnosis of haematopoietic malignancies. Detection of characteristic genetic abnormalities is extremely important in establishing the correct diagnosis and selecting the optimal treatment for the patient. In this paper we outline the problems of cytogenetic and molecular methods applied in the oncogenetic diagnostics. Moreover, we emphasize advantages and indicate limitations of these methods. Additionally, we present their practical use on the example of a comprehensive analysis of genetic changes in leukemic cells in a pediatric patient diagnosed with acute myeloid leukemia (AML). Simultaneous, proper interpretation of the results of all performed analysis – classical karyotype, fluorescence <I>in situ</I> hybridization (FISH), analysis of the expression of fusion genes and molecular karyotyping by microarrays, eventually led to the selection of optimal treatment which take into account the identified adverse genetic changes of the described patient.

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Valproic Acid Plus Retinoic Acid Induce Myeloid Differentiation in Chemotherapy-Resistant Acute Myeloid Leukemia Patients.

Blood ◽

10.1182/blood.v104.11.1805.1805 ◽

2004 ◽

Vol 104 (11) ◽

pp. 1805-1805 ◽

Cited By ~ 1

Author(s):

C. Nervi ◽

F. Lo Coco ◽

S. Minucci ◽

A. Careddu tech. ◽

R. Fiorini ◽

...

Keyword(s):

Valproic Acid ◽

Acute Myeloid Leukemia ◽

Retinoic Acid ◽

Plasma Levels ◽

Myeloid Leukemia ◽

Intensive Therapy ◽

Myeloid Differentiation ◽

Fish Analysis ◽

Leukemic Clone ◽

Acute Myeloid

Abstract The anti-epileptic drug valproic acid (VPA) acts as an inhibitor of histone deacetylases. In combination with retinoic acid (RA), VPA triggers myeloid differentiation of primary acute myeloid leukemia (AML) blasts in vitro. In vivo, VPA posses an antineoplastic activity as indicated by pre-clinical studies in murine models of leukemia, renal and lung metastatic tumors. Therefore, we have designed a phase II clinical study in which VPA was combined with RA (VPA-RA) in the AML treatment. Eigth chemotherapy-resistant or high risk AML patients not eligible for additional intensive therapy (median age 61.5 yrs), were treated at the Hematology Units of the Universities “La Sapienza” and “Tor Vergata” Rome-Italy. VPA (Depakin[Sanofi-Wintrop]) was administrated from day 1 to day 28, at the initial dosage of 10 mg/kg/die p.o. with dose escalation until optimal VPA plasma levels (80–110ug/ml). RA (Vesanoid [Roche]) at the dosage of 45 mg/m2 p.o./d, divided in two administrations, was added once the optimal VPA plasma levels were reached or at day 14 and continued until day 28. Four patients had a history of MDS, three patients had a FAB M0, M1 and M2 de novo AMLs, while the remaining case was a myeloid blast crisis (FAB M0) of a Ph+ve CML. Cytogenetic characterization in the other patients revealed normal karyotype in one case, a pseudodiploid [der(12)] in one, hyperdiploid (+8) in one, complex K with a 7q- alteration in one, while in the three remaining cases the karyotype was not evaluable. Pre-treatment leukemic infiltration ranged from 22% to 95%. VPA plasma level >60mg/ml was reached between 8 to 28 days (median 14.5 days). In three patients, VPA-RA treatment induced hyperleukocytosis (>50x 109/l) at day 16, 21 and 24, respectively, that was treated with chemotherapy (HU in two cases and low dose Ara-C in 1 case). Hematological improvement (≥50% decrease in packed red blood cell or platelet transfusion requirement) was observed in one case, a stable disease in five cases and disease progression in two cases. Peripheral blood and/or bone marrow samples were collected at day 0,3,7,14,21,28 for morphologic, immunophenotypic, cytogenetic and molecular studies. All patients showed features of myeloid-monocytic and/or erythroid differentiation of the leukemic clone, as revealed by morphologic, cytochemical, immunophenotypic analyses and by Q-RT-PCR of myeloid gene expression (GATA 1, MPO, CSF2Rb, etc.). Of note that high degree of myeloid differentiation correlated with early achievement of therapeutic VPA plasma levels and histone hyper-acetylation, as measured by immunocytochemistry and immunoblotting using antiacetylated histone H3 and H4 antibodies. Finally, differentiation of the leukemic clone was proven by FISH analysis showing the presence of the +8 and 7q- in maturing elements in patients whose leukemia blasts carried these cytogenetic lesions. The VPA-RA combination is a well tolerated treatment that induces phenotypic changes of the leukemic clone through chromatine remodelling. Further studies are needed to optimise this regimen with the aim of improving clinical response in leukemia patients.

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Mutations of the TP53 Gene Play an Important Role in the Pathogenesis of Acute Myeloid Leukemia with Complex Aberrant Karyotype.

Blood ◽

10.1182/blood.v104.11.63.63 ◽

2004 ◽

Vol 104 (11) ◽

pp. 63-63 ◽

Cited By ~ 9

Author(s):

Claudia Schoch ◽

Frank Dicker ◽

Alexander Kohlmann ◽

Sonja Rauhut ◽

Wolfgang Hiddemann ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Tp53 Mutation ◽

Direct Sequencing ◽

Tp53 Gene ◽

Single Amino Acid ◽

Comparative Genomic ◽

Fish Analysis ◽

Missense Mutations ◽

Acute Myeloid

Abstract Acute myeloid leukemia with complex aberrant karyotype is associated with the most unfavorable prognosis of all AML subtypes. Clinical behavior, complexity of karyotype abnormalities and the exponential increase of incidence with age shows similarities to the most common solid tumors. In the latter ones TP53 gene is the most frequently mutated gene identified so far. Therefore, we addressed the role of TP53 in AML with complex aberrant karyotype. In a first step we analyzed 360 AML cases with complex aberrant karyotype using conventional cytogenetics and 24 color FISH to resolve the chromosomal rearrangements in detail. In combination with data from comparative genomic hybridization for a subset of these patients (n=49) we identified 10 genomic regions frequently lost (5q14q33, 7q32q35, 12p13, 13q14, 16q22q24, 17p13, 18q21q22) or gained (11q23q25, 1p33p36, 8q22q24). In all 350 cases interphase FISH with a TP53 probe was performed. In 210 patients (60%) a loss of one TP53 allele was observed. 34 cases (22 with and 12 without TP53 loss) were further evaluated for p53 mutations using the Affymetrix p53 GeneChip assay. All mutations detected were verified by direct sequencing. TP53 mutations were detected in 21 of the 22 cases showing a loss of one TP53 allele (95%), while 9 of 12 cases (75%) without a TP53 loss showed a TP53 mutation on the microarray. Two mutations occurred in introns 4 and 7, while all others were located in exons (e4: n=2, e5: n=10, e6: n=3, e7: n=5, e8: n=8, e9: n=1). 22 mutations were missense mutations resulting in the substitution of a single amino-acid, while 4 were nonsense mutations. One small deletion and one insertion were detected. Four cases without mutations detected by microarray screening and 15 additional cases (9 with and 11 without TP53 loss detected with FISH) were further analyzed by DHPLC (WAVE, Transgenomics). In two of the four cases in which no mutation was detectable on the microarray mutations were found with DHPLC, which are most likely larger deletions not detectable with the p53 GeneChip assay. Furthermore, in 11 of the 15 additional cases a mutation was detected by DHPLC. Thus, in total 43 of 49 cases (88%) showed a TP53 mutation (27/30 (90%) with loss of one TP53 allele and 16/19 (84%) without loss of one TP53 allele). Three of the 6 cases in which no TP53 mutation was detected showed loss of one TP53 allele in FISH analysis. Taken together, only in 3 of 49 cases (6%) no alteration of TP53 was detected. In one of these cases an increased MDM2 expression was found using gene expression microarrays (U133A), another mechanism of inactiving TP53 function. In conclusion, the loss of normal TP53 function by loss of one allele and/or point mutation plays an important role in the pathogenesis of AML with complex aberrant karyotype and may be a major reason for chemoresistance in this prognostically most unfavorable AML subtype. TP53 alterations are detectable in more than 90% of cases with complex aberrant karyotype and are very rare in other AML subtypes. Therefore, we suggest to include the TP53 status in the definition of AML with complex aberrant karyotype.

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