Masayuki Kurata

The gene for interleukin-21 receptor is the partner of BCL6 in t(3;16)(q27;p11), which is recurrently observed in diffuse large B-cell lymphoma.

BCL6 translocation affecting the chromosomal band 3q27 can involve a number of non-immunoglobulin (non-IG) gene loci as partners. We report here that the gene for interleukin-21 receptor (IL-21R) is the partner of BCL6 in t(3;16)(q27;p11) translocation. The two breakpoints on 16p11 of a lymphoma cell line YM and case no. 1012 with diffuse large B-cell lymphoma, both of which carried t(3;16), were localized within the ∼27-kb intron 1 of IL-21R. As a result of t(3;16), the promoter region of IL-21R was substituted for the regulatory sequences of BCL6 in the same transcriptional orientation. Reverse transcriptase-mediated polymerase chain reaction revealed chimeric mRNA consisting of two non-coding exons 1a/1b of IL-21R and coding exons of BCL6 in both lymphoma cells. Fluorescence in situ chromosomal hybridization of YM metaphase cells revealed fusion signals that contained both the BCL6 and IL-21R sequences on the der(3)t(3;16) chromosome. IL-21R was actively transcribed in YM cells, while BCL6 that was under the control of the IL-21R promoter was only moderately expressed at the mRNA and protein level. We constructed expression plasmid of BCL6 that followed the promoter sequences of IL-21R. COS-7 cells transiently transfected with the plasmid expressed high level Bcl-6 protein and displayed nuclear staining with a characteristic punctate pattern by immunofluorescence, indicating that expression of BCL6 can be enhanced by t(3;16). This study added to the list of non-IG partners of BCL6 translocations a new class of gene, i.e. cytokine receptor gene, the expression of which is closely associated with lymphoid cells.

Fulminant fatal cardiotoxicity following cyclophosphamide therapy

A 59-year-old male with an abdominal mass that showed a diffuse large B cell lymphoma underwent extirpation of the tumor and chemotherapy. He subsequently received high-dose chemotherapy containing cyclophosphamide (1.5 g/m(2)/day x 2 days), followed by autologous peripheral blood stem cell transplantation. He developed congestive heart failure 5 days after administration of cyclophosphamide. His electrocardiogram showed extremely low voltage with ST segment change and echocardiogram showed diffusely increased left ventricular wall thickness, an increase in myocardial echogenicity, pericardial effusion, and generally decreased systolic function. Congestive heart failure progressed rapidly and he died the following day. Post-mortem examination of the heart revealed myocardial hemorrhage, yellowish brown pericardial effusion, and fibrinous pericarditis. His liver was atrophic and focal necrosis was observed histologically. Cyclophosphamide-induced cardiotoxicity occurred, even though the patient had both shown normal cardiac function before high-dose chemotherapy and had received a lower dose of cyclophosphamide. Concomitant administration of cytarabine might have affected his liver function and there might have been interaction between the drugs.

Excellent response of chemotherapy-resistant B-cell-type chronic lymphocytic leukemia with meningeal involvement to rituximab

A 70-year-old woman was diagnosed with B-cell-type chronic lymphocytic leukemia (B-CLL) in May 2001. Initial white blood cell (WBC) count was 37 × 109/l and most of the cells were mature small lymphocytes. Surface antigen analysis of these lymphocytes revealed positive reactions for CD19, 20, 25, 5, and λ-light chain. Despite her Rai stage-0 status, various treatments were ineffective, including cyclophosphamide; fludarabine; 6-mercaptopurine; a combination of vincristine, cyclophosphamide, prednisolone, and adriamycin; and etoposide. Her WBC count increased, ranging from 150 to 450 × 109/l, with marked splenomegaly, and symptoms of meningitis, such as headache, ophthalmalgia, hearing disturbance, and abnormal behavior, being manifested. The WBC count in the cerebrospinal fluid was elevated to 134/µl. The surface phenotype of these cells was identical to that of circulating lymphocytes, indicating meningeal involvement of leukemia, a rare complication in B-CLL. At the time of this WBC elevation, 24% of circulating lymphocytes had prominent nucleoli, indicating progression of the disease to CLL/prolymphocytic leukemia. Her symptoms disappeared after repeated intrathecal injections of methotrexate and dexamethazone. After four courses of treatment of the refractory B-CLL with rituximab, an anti-CD20 monoclonal antibody, the WBC count returned to normal levels and the splenomegaly disappeared. She is currently well, with sustained remission, as of April 2004.

Efficacy and tolerability of reduced-dose 21-day cycle rituximab and cyclophosphamide, doxorubicin, vincristine and prednisolone therapy for elderly patients with diffuse large B-cell lymphoma

Abstract Rituximab combined with cyclophosphamide, doxorubicin, vincristine and prednisolone (R-CHOP) is regarded as the first-line treatment for elderly patients with diffuse large B-cell lymphoma (DLBCL), but it is often necessary to reduce the dose or prolong the intervals between doses. In our center, dose reduction is performed with elderly patients with DLBCL on an individual basis: for patients in their 70s and 80s, the initial CHOP dose is reduced to 70% and 50%, respectively, and the subsequent doses are adjusted so that the patients receive, at 21-day intervals, the highest dose they can tolerate (reduced-dose R-CHOP21). To verify this practice, a retrospective analysis was performed. Between January 2004 and January 2011, 109 ≥ 70-year-old patients with DLBCL received reduced-dose R-CHOP21 with curative intent. The 2-year overall survival rates of the 70–79- and ≥ 80-year-old patients were 75.2% and 64.6%, respectively. Of 35 deaths, 20 were due to disease progression and five were related to treatment toxicity. Multivariate analysis revealed that an age of 75–79 years and an age of 80 years or older were associated with shorter survival. Given that many patients had poor performance status and comorbidities, reduced-dose R-CHOP21 may provide a reasonable balance between efficacy and tolerability for elderly patients with DLBCL.

Chronic eosinophilic Leukemia with the fip1l1-pdgfr± fusion gene in a patient with a history of combination chemotherapy

Hypereosinophilic syndrome (HES) was diagnosed in December 2000 in a 43-year-old man on the basis of persistent eosinophilia (11.7 x 109/L) and a normal karyotype of the bone marrow cells. He had developed intra-abdominal non-Hodgkin’s lymphoma and in 1992 had received 3 courses of combination chemotherapy with doxorubicin (Adriamycin), cyclophosphamide, vincristine, methotrexate, bleomycin, and prednisolone. The patient was orally given prednisolone (10 mg/day) and cyclophosphamide (50 mg/day) as HES treatment without a subsequent improvement of the eosinophilia. In May 2003, anemia (hemoglobin, 7.9 g/dL) and thrombocytopenia (65 x 109/L) manifested with progressive eosinophilia (21.0 ×109/L) and a small number of blasts. The patient became febrile and was admitted in July 2003. Cytogenetic reexamination of the bone marrow cells disclosed the deletion of 4q12, indicating the presence of a fusion of the Fip1-like 1 (FIP1L1) gene to the plateletderived growth factor receptor α (PDGFRα) gene and consequently the clonal nature of his hematopoietic cells. DNA sequence analysis demonstrated that the breakpoints of the FIP1L1 and PDGFRα genes were present in exon 9 and exon 12, respectively. Treatment with imatinib mesylate (300 mg/day) promptly brought about complete remission. Although a number of similar eosinophilic cases have been reported, our patient may be the first such patient with a history of chemotherapy.

Successful allogeneic bone marrow transplantation for myelodysplastic syndrome complicated by severe pulmonary alveolar proteinosis

Pulmonary alveolar proteinosis (PAP) is a rare disorder characterized by the abnormal accumulation of alveolar surfactant protein in alveolar spaces. We report herein a rare case of myelodysplastic syndrome (MDS-RAEB) complicated by severe PAP, and successful allogeneic bone marrow transplantation (BMT) for both disorders. An unrelated BMT was planned for a 48-year-old male with advanced MDS-RAEB. Just before the initiation of the conditioning regimen for unrelated BMT in March 2007, he developed dyspnea. A diagnosis of PAP was made based on findings of chest X-ray, CT scanning, and the fluid obtained by bronchoalveolar lavage. To improve his dyspnea and improve BMT safety, whole lung lavage (WLL) was performed twice, with the partial improvement of PAP. Unrelated allogeneic BMT was performed in September 2007. We had to perform a third WLL because of the worsening of PAP on day 26 after BMT. Despite many infectious complications after BMT, GVHD was relatively mild. PAP had almost disappeared 6 months after BMT. He was well with favorable hematopoiesis 20 months after the BMT without any specific treatment. There has been no report of an MDS patient with PAP in whom 3 WLL procedures were performed before and after allogeneic BMT.

Hepatosplenic αβ T cell lymphoma

A 32-year-old male with chronic hepatitis B was admitted to a hospital with cellulitis in the right leg in September 2006. Pancytopenia, hepatosplenomegaly, and systemic superficial lymph node swelling were noted, and he was referred to our hospital. He developed fever and liver dysfunction in June 2007 and underwent a splenectomy. His pancytopenia subsequently improved. A pathologic diagnosis of hepatosplenic αβ T cell lymphoma was made by examining spleen tissue and biopsy specimens of the liver and mesenteric lymph node. He had stage IVB disease because neoplastic T cells were noted in the bone marrow. The response of the lymphoma to conventional chemotherapy including the CHOP (cyclophosphamide, adriamycin, vincristine, prednisolone) and DeVIC (dexamethasone, etoposide, ifoshamide, carboplatin) regimens was poor and transient. A partial remission was obtained with an ESHAP (etoposide, cisplatin, cytarabine, methylprednisolone) regimen. Therefore, we planned a bone marrow transplantation (BMT) from an HLA-haploidentical sibling donor. He was moved to the Department of Hematology, Hyogo Medical College, to receive this BMT as part of a clinical trial. During the conditioning procedure for the transplantation, however, he died of septicemia. Since hepatosplenic αβ T cell lymphoma is very rare with only 23 reported cases to date, herein we report this case and discuss the therapeutic strategy.

Impact of occult bone marrow involvement on the outcome of rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisone therapy for diffuse large B-cell lymphoma

Abstract We assessed the prognostic impact of occult bone marrow involvement, determined by flow cytometry and/or polymerase chain reaction, in a population of 117 consecutive patients with newly diagnosed diffuse large B-cell lymphoma (DLBCL) treated with rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP). Twenty-four (20.5%) had morphologically diagnosed and 16 (13.7%) had occult marrow involvement, and 77 (65.8%) had no marrow involvement. Although the pretreatment characteristics of the negative or occult marrow involvement group were similar, severe hematological toxicity after R-CHOP was more frequent in the occult marrow involvement group. Progression-free survival (PFS; p = 0.015) and overall survival (OS; p = 0.035) for the occult marrow involvement group were significantly shorter than those for the negative group, and were comparable to those of the morphologic marrow involvement group, independent of the International Prognostic Index score for PFS. Occult bone marrow involvement predicts severe hematological toxicity and negatively impacts on the PFS and OS of R-CHOP therapy.

Refractory de novo myeloid sarcoma: a case report and therapeutic strategy based on bone marrow minimal residual disease

Myeloid sarcoma (MS), which usually develops concurrently with acute myeloid leukemia (AML), is an extramedullary tumor that consists of immature myeloid cells. Clinically, the incidence of MS in AML is 3–5% [1, 2], and the association of MS is linked to a poor prognosis [3]. De novo MS is characterized by leukemic tumor formation without the involvement of bone marrow and peripheral blood. Patients receiving radiotherapy or incorrect chemotherapy develop AML at a median time of 7 and 12 months, respectively, and show an extremely poor prognosis after leukemic transformation [4]. Thus, systemic chemotherapy effective against AML, which lowers the rate of leukemic transformation from 71 to 41%, is essential for the treatment of de novo MS [5]. A 17-year-old male was referred to our hospital in April 2007, because of small tumors in the left neck and supraclavicular fossa. A CT scan showed several separated tumors in the neck and mediastinum, one being 4 9 2 cm in the superior mediastinum and the others around 1.5 cm. The histologic picture of a biopsied left supraclavicular lymph node demonstrated that the normal architecture had been destroyed and completely replaced by middle-sized immature myeloid cells (Fig. 1), which expressed T-cell markers including CD2 and CD7 in addition to CD13, CD33, CD38, CD43, cytoplasmic myeloperoxidase, and HLA-DR on flow cytometry. Chromosomal analysis of the specimen revealed an abnormal karyotype of 46,XY,i(17)(q10) in 5 of 7 dividing cells. Blood and marrow cells did not contain abnormal cells as evaluated by flow cytometry and G-banding. A diagnosis of de novo MS was made. After 2 courses of intensive chemotherapy effective against AML, a complete remission (CR) of MS was achieved as evaluated by CT scan. Two months after the initial therapy, however, we detected a small population of myeloblasts (0.05% of all nucleated cells), which aberrantly expressed CD2 and CD7 in the bone marrow on flow cytometry, regardless of the disappearance of MS tumors. Thus, we regarded CD2/CD13 myeloblasts as MRD in the bone marrow. Subsequently, 3 courses of consolidation therapy including high-dose cytarabine were performed; however, MRD in the bone marrow persisted (0.05– 0.20%). Although we planned allogeneic HSCT, overt AML rapidly developed during the recovery phase after the third consolidation chemotherapy. The leukemia was highly refractory to any treatment including the regimens for acute lymphoblastic leukemia after December 2007. Chromosomal analysis of the bone marrow cells showed an abnormal karyotype of 46,XY,t(1;11)(p10;q10),add(4) (q21),i(17)(q10) in 8 of 20 dividing cells in addition to that D. Inoue (&) Y. Nagai T. Kimura S. Shimoji M. Mori K. Togami S. Tabata M. Kurata A. Matsushita K. Nagai T. Takahashi Department of Hematology and Clinical Immunology, Kobe City Medical Center General Hospital, 4-6 Minatojima nakamachi, Chuo-ku, Kobe 650-0046, Japan e-mail: daichi@kcgh.gr.jp

Multiple granulocytic sarcomas in essential thrombocythemia

A 59-year-old woman was diagnosed with essential thrombocythemia in 1988 and had been treated with hydroxyurea, mitobronitol, busulfan, and ranimustine, in that order. Hepatosplenomegaly, low-grade fever, and body weight loss manifested, and a few blasts were noted in the peripheral blood studied in March 2002. A biopsied specimen of the bone marrow showed myelofibrosis but not a leukemia in August 2004. An abnormal karyotype with der(1;13) appeared for the first time. She was treated with low-dose prednisolone. In January 2005, she experienced left hip joint pain, and magnetic resonance scanning showed a tumoral lesion in the femoral head. Histological diagnosis of the biopsied mass revealed that it was a granulocytic sarcoma, and radiotherapy was performed. In April 2005, bone scintigraphy showed multiple lesions. She became febrile and red blood cell transfusion-dependent with hepatosplenomegaly and a small number of circulating blasts. Intravenous cytarabine (low dose) and etoposide relieved the fever and hepatosplenomegaly; however, she developed a pathologic fracture of the right humerus.An additional karyotypic abnormality (7q22 deletion) was noted. She subsequently died of infection. Granulocytic sarcoma is very rare in essential thrombocythemia, and this patient may be the first reported case of essential thrombocythemia that developed multiple lesions and a pathologic fracture without transformation to overt leukemia.