Shinpei Furusawa

Expression of the NUP98/HOXA9 fusion transcript in the blast crisis of Philadelphia chromosome‐positive chronic myelogenous leukaemia with t(7;11)(p15;p15)

The t(7;11)(p15;p15) translocation is a recurrent aberration observed in acute myeloblastic leukaemia (AML) and chronic myelogenous leukaemia (CML). It has been shown that the NUP98 gene at 11p15 is fused with the HOXA9 gene at 7p15 in AML with t(7;11). We report the first case with CML expressing the NUP98/HOXA9 fusion transcript. A 27‐year‐old Japanese man was initially diagnosed as in the chronic phase of Philadelphia‐positive CML. At the diagnosis of myeloid blast crisis, the karyotype evolved to 46, XY, t(7;11)(p15;p15), t(9;22)(q34;q11). Reverse transcriptase polymerase chain reaction identified the NUP98/HOXA9 transcript, suggesting that the NUP98/HOXA9 fusion protein could play a critical role in the progression to blast crisis.

Poor response to intensive chemotherapy in de novo acute myeloid leukaemia with trilineage myelodysplasia

Summary. The findings of morphologically dysplastic features in haemopoietic cells in de novo acute myeloid leukaemia (AML) has been named AML with trilineage myelodysplasia (AML/TMDS). We analysed the clinical data, karyotypes, and treatment outcomes of 230 de novo AML patients treated with the Japan Adult Leukaemia Study Group AML‐87 protocol. 40 (17%) patients had AML/TMDS. Platelet count was significantly higher (F=0·006) and bone marrow blasts were fewer (P=0·01) in the AML/TMDS group than in the AML without TMDS. Abnormal karyotype was shown in 12/30 patients (40%) analysed.

Phase I/II study of humanized anti-CD33 antibody conjugated with calicheamicin, gemtuzumab ozogamicin, in relapsed or refractory acute myeloid leukemia: final results of Japanese multicenter cooperative study

The primary objective of this study was to investigate the tolerability, efficacy and pharmacokinetic profile of gemtuzumab ozogamicin (GO) in patients with relapsed and/or refractory CD33-positive acute myeloid leukemia (AML). Patients received 2-h infusions of GO twice with an interval of approximately 14 days. Tolerability was assessed using the National Cancer Institute Common Toxicity Criteria Version 2.0. Samples for pharmacokinetics were taken on day 1 and day 8 of the first treatment cycle. The dose was increased stepwise and, in each cohort, patients were treated at the same dose. Forty patients, median age 58 years (range 28–68) were treated; 20 and 20 patients were enrolled to the phase I and II parts, respectively. In the phase I part, dose-limiting toxicities (DLTs) were hepatotoxicities, and the recommended dose was established as 9 mg/m2 given as two intravenous infusions separated by approximately 14 days. The pharmacokinetic study revealed that Cmax and AUC were equivalent to those of non-Japanese patients. In the phase II part, complete remission was observed in 5 patients, and one patient had complete remission without platelet recovery. Four of these 6 in remission and one in the phase I are long-term survivors (alive for at least 44 months). GO is safe and effective as a single agent among Japanese CD33-positive AML patients. Remission lasted longer in a subset of patients than in non-Japanese patients in earlier studies. Further studies of this agent are warranted to establish standard therapy.

Triple Philadelphia chromosomes with major-bcr rearrangement in hypotriploid erythroleukaemia.

Abstract: The Philadelphia (Ph) chromosome is observed in approximately 1% of patients with acute myeloblastic leukaemia (AML), especially subtypes M1 and M2 in the French–American–British classification. We describe here a cytogenetic and molecular investigation of a rare case with Ph‐positive AML M6 (erythroleukaemia). A 63‐yr‐old woman was diagnosed as having erythroleukaemia. Leukaemic cells were positive for CD4 and CD7 as well as CD13, CD33, CD34 and HLA‐DR. They were analyzed by G‐banding, fluorescence in situ hybridization (FISH), Southern blot and reverse transcriptase polymerase chain reaction analyses. The karyotypes at diagnosis were as follows: 61, XX, −X, −1, −2, −3, −4, −5, −7, t(9;22)(q34;q11)×2, −15, −16, −17, −18, +19, +21, +22 [3]/61, idem, −22, +der(22)t(9;22) [36]. FISH with BCR/ABL probes showed that 39% and 57% of interphase nuclei had double and triple BCR/ABL fusion signals, respectively. Chromosome analysis in complete remission showed a normal karyotype in all 20 metaphases, confirming the diagnosis as Ph positive‐acute leukaemia. FISH at relapse showed that 92% of interphase nuclei had triple fusion signals. Rearrangement of major breakpoint cluster region (M‐bcr) in the BCR gene and coexpression of p210‐type (b2a2) and p190‐type (e1a2) BCR/ABL fusion transcripts due to alternative splicing were also detected. We conclude that clonal evolution from double to triple Ph chromosomes may be implicated in the disease progression. Considering other two reported cases, Ph‐positive erythro‐leukaemia appears to be correlated with coexpression of myeloid/T‐lymphoid markers and hyperdiploidy with double or triple Ph chromosomes, although breakpoints in the BCR gene are heterogenous.

Translocation (14;19)(q32;q13) detected by spectral karyotyping and lack of BCL3 rearrangement in CD5-positive B-cell lymphoma associated with hemophagocytic syndrome.

It has been shown that some cases of B-cell non-Hodgkin lymphoma associated with a hemophagocytic syndrome (B-LAHS) have chromosomal abnormalities at 14q32 or 19q13. We report here a 64-year-old woman with B-LAHS and a complex karyotype including add(14)(q32). We applied spectral karyotyping and revealed that the add(14)(q32) was derived from a der(14)t(14;19)(q32;q13). However, rearrangement of the BCL3 gene at 19q13 could not be detected by Southern blot analysis. Our results indicate that the translocation involving 19q13 may be one of the recurrent aberrations in B-LAHS and that the molecular mechanism of t(14;19)(q32;q13) in B-LAHS appear to be different from that observed in chronic lymphocytic leukemia.

Analysis of internal deletions within the BCL6 gene in B‐cell non‐Hodgkin's lymphoma

The BCL6 gene is frequently altered by chromosomal translocations and/or point mutations at its 5′ non‐coding portion in B‐cell non‐Hodgkins lymphoma (B‐NHL). We analysed submicroscopic structural alterations of the BCL6 gene which had arisen from internal deletion in four cases with B‐NHL and found that these deletions overlapped at the 280 bp region in the first intron. In electrophoretic mobility shift assay, nuclear extracts prepared from various cell lines were shown to bind to a fragment from this commonly deleted region. Our results suggest that deregulation of BCL6 expression would be caused by loss of this putative protein‐binding sequence in some B‐NHL cases.

Clinical effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on various types of neutropenia including cyclic neutropenia

Recombinant human granulocyte colony-stimulating factor (rhG-CSF) was investigated for its clinical efficacy in the treatment of various types of neutropenia (3 cases with idiopathic neutropenia of suspected drug induction, 5 cases with idiopathic neutropenia of other origin, and 2 cases with cyclic neutropenia). Treatment with glycosylated rhG-CSF produced in the Chinese Hamster Ovary cells at dose levels of 2–5μg/kg/day caused rapid increases of neutrophil counts associated with an improvement of the infection. In cyclic neutropenia patients, marked reduction in the duration of the neutropenic period was observed with rhG-CSF administration started before the period. Intercurrent stomatitis, which occurred in 1 patient, was markedly milder as compared to a previous episode which occurred during an untreated neutropenic period.The treatment of rhG-CSF was well tolerated and no adverse events were observed, nor was there any detectable anti-rhG-CSF antibody in any patients studied; hence the clinical use of rhG-CSF is considered to be safe.These results suggest beneficial effects of rhG-CSF on the recovery of neutrophil counts in cyclic and other types of idiopathic neutropenias, as well as for the treatment of neutropenia-associated infection.

Identification of Surface Molecules on Eosinophils and Lymphocytes in Blood from Patients with Eosinophilia

Surface molecules on eosinophils and lymphocytes in blood from patients with hypereosinophilic syndrome (HES), Kimuras disease and normal volunteers were examined. In all 3 patients with HES, CD54-positive eosinophils were increased and in some patients with HES and Kimuras disease HLA-DR-positive eosinophils were increased. Additionally, CD11b-positive, CD16-positive, CD25-positive, CD54-positive, CD69-positive and HLA-DR-positive lymphocytes were increased in some of these patients.

Vesnarinone-Induced Granulocytopenia: Incidence in Japan and Recommendations for Safety

Vesnarinone (OPC‐8212) is a new positive inotropic agent that augments myocardial contractility. A recent multi‐center randomized trial in the United States demonstrated that 60 mg/day of vesnarinone significantly reduced morbidity and mortality and improved quality of life in patients with symptomatic chronic heart failure. Vesnarinone, however, is also known for its propensity to cause granulocytopenia. In search of effective safety measures against this side effect, data have been collected in Japan as part of the post‐marketing surveillance of this drug. This article reviews the results of this post‐marketing surveillance and other works available to date, including an illustrative case report, and presents measures that should be taken with regard to safety during treatment with vesnarinone. Vesnarinone‐induced granulocytopenia has appeared in relatively early stages of vesnarinone therapy, and characteristically results in a rapid decrease in granulocyte count. Hematologic monitoring should be performed at least once a week during the initial 16 weeks of vesnarinone therapy. Granulocyte colony‐stimulating factor may contribute to recovery from severe granulocytopenia, although it should be used carefully because of its potential to cause adult respiratory distress syndrome.

GM-CSF and Eosinophil Chemotactic Factors in an Acute Lymphoblastic Leukemia Patient with Eosinophilia

We describe a patient with common acute lymphoblastic leukemia associated with blood and cerebrospinal fluid (CSF) eosinophilia. Serum obtained at onset and conditioned medium prepared from T cells obtained at remission stimulated with interleukin-2 contained eosinophil colony stimulating activity (Eo-CSA), which was confirmed to be predominantly GM-CSF. Leukemic cell conditioned medium and serum obtained at remission contained no Eo-CSA. The CSF contained increased eosinophil chemotactic activity, however, this factor was not identified.