Makoto Ibata

Efficacy of folinic acid in preventing oral mucositis in allogeneic hematopoietic stem cell transplant patients receiving MTX as prophylaxis for GVHD

As the safety of folinic acid administration and its efficacy for reducing the toxicity of MTX remain controversial, we assessed the effect of folinic acid administration after MTX treatment for GVHD prophylaxis on the incidence of oral mucositis and acute GVHD. We retrospectively analyzed data for 118 patients who had undergone allogeneic hematopoietic SCT and had received MTX for GVHD prophylaxis. Multivariate analysis showed that systemic folinic acid administration significantly reduced the incidence of severe oral mucositis (odds ratio (OR)=0.13, 95% confidence interval (CI) 0.04–0.73, P=0.014). There was also a tendency for a lower incidence of severe oral mucositis in patients who received folinic acid mouthwash (OR=0.39, 95%CI 0.15–1.00, P=0.051). No significant difference was observed in the incidence of acute GVHD between patients who received systemic folinic acid administration and those who did not (P=0.88). Systemic folinic acid administration and mouthwash appear to be useful for reducing the incidence of severe oral mucositis in patients who have received allogeneic hematopoietic SCT using MTX as GVHD prophylaxis.

FIP1L1 presence in FIP1L1-RARA or FIP1L1-PDGFRA differentially contributes to the pathogenesis of distinct types of leukemia

FIP1-like 1 (FIP1L1) is associated with two leukemogenic fusion genes: FIP1L1-retinoic acid receptor alpha (RARA) and FIP1L1-platelet-derived growth factor receptor alpha (PDGFRA). Analyses of a series of deletion mutants revealed that the FIP1 motif in FIP1L1-RARA plays a pivotal role in its homodimerization and transcriptional repressor activity. However, in FIP1L1-PDGFRA, the C-terminal PDGFRA portion possesses the ability of forming a homodimer by itself, making FIP1L1 dispensable for constitutive activation of this kinase. Both the full-length and the C-terminal PDGFRA portion of FIP1L1-PDGFRA could transform the IL-3-dependent hematopoietic cell line, BAF-B03. Moreover, when either the full-length or the C-terminal PDGFRA portion of FIP1L1-PDGFRA was introduced in these cells, they grew in the absence of IL-3. The cells having the C-terminal PDGFRA portion of FIP1L1-PDGFRA, however, were partially IL-3 dependent, whereas the cells having the full-length FIP1L1-PDGFRA became completely IL-3 independent for their growth. Taken together, these results show that FIP1L1 differentially contributes to the pathogenesis of distinct types of leukemia.

Serum macrophage migration inhibitory factor (MIF) levels after allogeneic hematopoietic stem cell transplantation

Macrophage migration inhibitory factor (MIF) may play an important role in the pathogenesis of acute graft‐versus‐host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo‐HSCT), as MIF plays an important role to regulate the production of tumor necrosis factor‐α (TNF‐α), one of the inflammatory cytokines which induces and exacerbates aGVHD. We examined the association between serum MIF levels and aGVHD vs. chronic GVHD (cGVHD) in allo‐SCT patients in this study. We found a significant increase in the peak serum MIF (14.46 ng ± 1.47 ng/ml) at onset in patients that developed aGVHD (n = 23, P = 0.009). We also found that mean serum MIF levels in patients who developed extensive type cGVHD within 6 months (12.58 ± 2.18 ng/ml, n = 13) were significantly higher than MIF levels before allo‐HSCT (7.86 ± 1.17 ng/ml, n = 19, P = 0.04). Therefore, we speculated that serum MIF levels increase during the active phase of both aGVHD and cGVHD.

Abnormal expansion of naïve B lymphocytes after unrelated cord blood transplantation : a case report

A 33-year-old woman underwent unrelated cord blood transplantation (U-CBT) for myelodysplastic syndrome (MDS)-related secondary AML. She showed impressive increases in the number of CD19+ B cells in bone marrow and CD19+27−IgD+ B cells in peripheral blood from about 1 month to 3 months after U-CBT. The serum level of IL-6 temporarily increased after transplantation, and this increase seemed to be correlated with the expansion of CD19+ B cells. Although, compared with BMT, little is known about the kinetics of hematological and immunological reconstitution in U-CBT, there was initial B-cell recovery after CBT as some described. This B cell recovery may be associated with a high number of B-cell precursors present in cord blood (CB). The phenomenon of naïve B lymphocyte expansion that we found might be associated with a high number of B-cell precursors present in CB.

Sequential chemotherapy and myeloablative allogeneic hematopoietic stem cell transplantation for refractory acute lymphoblastic leukemia

The prognosis of patients receiving allogeneic hematopoietic stem cell transplantation (HSCT) for refractory acute lymphoblastic leukemia (ALL) is very poor. To improve survival rates, we attempted to intensify the conditioning regimen with daunorubicin, vincristine, prednisolone, medium-dose etoposide, cyclophosphamide, and total body irradiation (DNR/VCR/PSL plus medium-dose VP/CY/TBI). Four patients in relapse or induction failure of B-precursor ALL without other complications underwent allogeneic HSCT. Initially, chemotherapy comprising DNR 60 mg/m2 for 3 days, VCR 1.4 mg/m2 for 1 day, and PSL 60 mg/m2 for 3 days was administered, which was followed by medium-dose VP/CY/TBI; some modifications were made for individual patients. All patients achieved engraftment and complete remission after HSCT. Regimen-related toxicities were tolerable and no patient died within 100 days. Two patients were alive without disease on days 563 and 1,055. The third patient relapsed on day 951, while the fourth died on day 179 without disease. Our results indicate that intensified myeloablative HSCT should be considered for patients with refractory ALL.

The 8p11 myeloproliferative syndrome owing to rare FGFR1OP2-FGFR1 fusion

To the Editor: The 8p11 myeloproliferative syndrome (EMS) is an aggressive neoplasm associated with chromosomal abnormalities involving the fibroblast growth factor receptor 1 (FGFR1) tyrosine kinase gene on chromosome 8p11–12. A 43-yr-old man was referred owing to progressive lymphoadenopathy and eosinophilia. Laboratory data showed an elevated white blood cell count with a markedly elevated eosinophil count and no circulatory blast (WBC: 36.5 · 10 ⁄L, eosinophils: 19.5%). The lactate dehydrogenase level was elevated (LDH 478 U ⁄L). Biopsy of a cervical lymph node revealed T-cell lymphoblastic lymphoma. Bone marrow aspiration revealed hypercellular marrow with markedly increased eosinophils (Fig. 1A). Chromosome analyses of the bone marrow and lymph node specimen were both initially interpreted as 46,XY [20]. Because of the unique presentation of concurrent T-cell lymphoblastic lymphoma and eosinophilia, we were suspected that the patient had a cryptic FGFR1 abnormality despite the normal karyotype. To confirm cryptic cytogenetic abnormality, we sent a sample for spectral karyotype fluorescence in situ hybridisation (SKY-FISH) analysis (SRL, Tokyo, Japan). SKYFISH revealed an obvious chromosomal abnormality, which could read 46,XY,t(8;12)(p11;p12) [5] (Fig. 1B). A literature search revealed one previous report with an

Ciprofloxacin inhibits lipopolysaccharide-induced toll-like receptor-4 and 8 expression on human monocytes derived from adult and cord blood

Dear Editor, Lipopolysaccharide (LPS) interacts with immune cells by binding to CD14 molecules and then interacts with toll-like receptor (TLR)-4 on monocytes and dendritic cells [1–3]. On the other hand, TLR8 recognizes viral single-stranded RNA (ssRNA) [4]. Therefore, TLR4 is involved in immunological events during bacterial infection, and TLR8 is involved in immunological events during viral infection. Ciprofloxacin (CIP), a fluorinated 4-quinolone, is one of the quinolone antibiotics. CIP reduces the population of intestinal bacteria and influences the development of acute graft-vs-host disease (GVHD) after allogeneic bone marrow transplantation for hematological malignancies [5]. In addition, CIP has modulatory actions on immune and inflammatory responses, such as inhibition of the production of tumor necrosis factor (TNF)-α in LPS-stimulated peripheral blood mononuclear cells (PBMC) [6]. The mechanisms underlying the effects of quinolones on immune modulation might depend on the regulation of intracellular cAMP and NF-kB. CIP induces the production of prostaglandin E2 and increases intracellular cAMP levels. A hyperresponse to LPS by monocytes induces symptoms of septic shock, and CIP might therefore contribute to the attenuation of these symptoms after severe infection [7]. Cord blood contains more naive T cells and Treg precursor cells than adult blood [8]. In addition, it has been reported that there is a profound defect in IL-12 (p70) synthesis and an increased release of IL-10 in cord blood exposed to LPS compared to those in adult blood [9]. These facts might be relevant to the increased vulnerability of human newborns to intracellular pathogens and the low incidence of GVHD after cord blood transplantation (CBT). In this study, we analyzed TLR4, TRL8, ICAM-1, and LFA-1 expression on the surface of CD14-positive monocytes and TLR4, TLR8, and TNF-α mRNA expression after LPS stimulation with CIP using adult PBMC and cord blood mononuclear cells (CB). Normal adult PBMC and cord blood units were obtained from healthy donors with informed consent from Hokkaido Red Cross Blood Center Sapporo. Cells (1×10/ml) were cultured for 3 days with 10 μg/ml LPS (Escherichia coli 055: B5, EMD Bioscience, La Jolla, CA, USA) in RPMI-1640 with 10% fetal calf serum in the presence or absence of 50 μg/ ml CIP (kindly provided by Bayer Yakuhin, Osaka, Japan). After 3 days of culture, cells were harvested and stained with FITC-conjugated anti-TLR4 (HTA125; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-LFA-1 (HI111; Becton Dickinson, San Diego, CA, USA), and anti-ICAM-1 (84H10; Immunotech, Marseille, France) monoclonal antibody (mAb) and with FITC-conjugated goat anti-mouse Ig G1 (STAR81F; Serotec, Oxford, UK) as a secondary antibody for anti-TLR8 (44C143; IMGENEX, San Diego, CA, USA) mAb and PEconjugated anti-CD14 (MoP9; Becton Dickinson) mAb. Mean fluorescence intensity (MFI) for TRL4, TRL8, ICAM-1, and LFA-1 on CD14-positive monocytes was analyzed using a fluorescence-activated cell sorter (FACS) Ann Hematol (2008) 87:229–231 DOI 10.1007/s00277-007-0363-x

Hyponatremia after administration of conditioning regimen in myelodysplastic syndrome with empty sella after glandula pituitaria surgery

Dear Editor, A 51-year-old woman who had developed myelodysplastic syndrome with an excess of blast [myelodysplastic syndrome (MDS), refractory anemia with excess blast (RAEB)] was admitted to our institution. She obtained complete remission after an administration of a low dosage of cytarabine. Then, she was administered busulfan (BU, 4 mg/kg for 2 days), fludarabine (FLU, 30 mg/m for 6 days), and total body irradiation (TBI, 2 Gy twice a day) as a conditioning regimen for unrelated stem cell transplantation (SCT) from a human leukocyte antigen (HLA) completely matched donor. Hyponatremia developed after the administration of BU, and the patient developed general fatigue. Her serum sodium level had fallen to 118 mEq/l (normal range from 135 to 147 mEq/l), plasma osmolarity was 240 mOsm/kg/H2O (normal range from 280 to 300 mOsm/kg/H2O), urine osmolarity was 502 mOsm/kg/l, and antidiuretic hormone (ADH) level was 2.3 pg/ml (normal range from 0.3 to 3.5 pg/ml) without edema. These facts suggested that an inappropriate secretion of ADH (SIADH) had developed after the conditioning regimen. After treatment with hypertonic saline infusion and minocycline hydrochloride (MINO), her hyponatremia gradually recovered. On the fourth day of the conditioning regimen, her serum sodium level recovered to 138 mEq/l. Magnetic resonance imaging (MRI) of the glandula pituitaria only showed neurohypophysis (lobus posterior) due to the previously performed Hardy’s operation for her pituitary adenoma. Adrenocorticotropic hormone (ACTH), thyroid-stimulating hormone (TSH), and adrenal hormone were all within normal ranges without hormone therapy support before SCT. Inappropriate secretion of ADH (SIADH) is characterized by water retention, hyponatremia, and central nervous system symptoms. It is induced by several factors including central nervous disorders, endocrine diseases, paraneoplastic syndromes, and such anticancer agents as vincristine and cyclophosphamide [1–4]. BU and FLU have never been reported as causes of SIADH. Our patient did not have any viral infection or disease recurrence, and only the administration of the conditioning regimen was evident after the development of SIADH. Kobayashi et al. reported hyponatremic patients after stem cell transplantation. They showed that being younger than 4 years old was a risk factor for hyponatremia after SCT. They suspected the release to be causative for the development of SIADH, especially in association with acute graft-versus-host disease (GVDH) [5]. But our case was an adult who developed SIADH before SCT. BU is transported to cerebrospinal fluid [6]. FLU effectively treats the invasion of the sanctuary space by chronic lymphocytic leukemia [7, 8], showing that FLU might also pass to the cerebrospinal fluid. Therefore, BU as Ann Hematol (2007) 86:843–844 DOI 10.1007/s00277-007-0291-9

Chimerism and T-cell receptor repertoire analysis after unrelated cord blood transplantation with a reduced-intensity conditioning regimen following autologous stem cell transplantation for multiple myeloma.

A 65‐year‐old Japanese male was diagnosed as multiple myeloma with Bence Jones kappa type, clinical stage IIIA. His disease status reached partial remission after chemotherapy. Thereafter, he received tandem transplantation, consisting of high‐dose chemotherapy with autologous stem cell transplantation (ASCT), followed by unrelated cord blood transplantation (U‐CBT). U‐CBT with a reduced‐intensity conditioning regimen (RI‐CBT) was performed in August 2003. HLA mismatch between the patient and the CBT donor was present at two serological loci (B and DR). A total nucleated CBT cell dose of 2.45 × 107/kg body weight was infused on day 0. Graft‐vs.‐host disease (GVHD) prophylaxis consisted of cyclosporine A and short‐term methotrexate. Neutrophil engraftment (>0.5 × 109/l) was obtained on day 46. He developed positive cytomegalovirus antigenemia, grade II acute GVHD involving skin and liver, varicella–zoster virus infection, septic shock, hemorrhagic cystitis caused by adenovirus and acute hepatitis B virus infection after U‐CBT. We retrospectively analyzed T‐cell receptor (TCR) repertoire diversity and found that TCR repertoire diversity decreased continuously after U‐CBT. Therefore, low‐TCR repertoire diversity in this patient appears to be associated with various infections caused by immunodeficiency.

Leukemogenic kinase FIP1L1-PDGFRA and a small ubiquitin-like modifier E3 ligase, PIAS1, form a positive cross-talk through their enzymatic activities

Title A leukemogenic kinase, FIP1L1-PDGFRA, and a SUMO E3 ligase, PIAS1, form a positive crosstalk via their enzymatic activities [an abstract of dissertation and a summary of dissertation review] Author(s) 井端, 淳 Citation 北海道大学. 博士(医学) 甲第12530号 Issue Date 2017-03-23 Doc URL http://hdl.handle.net/2115/65866 Rights(URL) http://creativecommons.org/licenses/by-nc-sa/2.1/jp/ Type theses (doctoral abstract and summary of review) Note 配架番号:2271 Additional Information There are other files related to this item in HUSCAP. Check the above URL. File Information Makoto_Ibata_abstract.pdf (論文内容の要旨)Fusion tyrosine kinases play a crucial role in the development of hematological malignancies. FIP1L1‐PDGFRA is a leukemogenic fusion kinase that causes chronic eosinophilic leukemia. As a constitutively active kinase, FIP1L1‐PDGFRA stimulates downstream signaling molecules, leading to cellular proliferation and the generation of an anti‐apoptotic state. Contribution of the N‐terminal FIP1L1 portion is necessary for FIP1L1‐PDGFRA to exert its full transforming activity, but the underlying mechanisms have not been fully characterized. We identified PIAS1 as a FIP1L1‐PDGFRA association molecule by yeast two‐hybrid screening. Our analyses indicate that the FIP1L1 portion of FIP1L1‐PDGFRA is required for efficient association with PIAS1. As a consequence of the association, FIP1L1‐PDGFRA phosphorylates PIAS1. Moreover, the kinase activity of FIP1L1‐PDGFRA stabilizes PIAS1. Therefore, PIAS1 is one of the downstream targets of FIP1L1‐PDGFRA. Moreover, we found that PIAS1, as a SUMO E3 ligase, sumoylates and stabilizes FIP1L1‐PDGFRA. In addition, suppression of PIAS1 activity by a knockdown experiment resulted in destabilization of FIP1L1‐PDGFRA. Therefore, FIP1L1‐PDGFRA and PIAS1 form a positive cross‐talk through their enzymatic activities. Suppression of sumoylation by ginkgolic acid, a small molecule compound inhibiting a SUMO E1‐activating enzyme, also destabilizes FIP1L1‐PDGFRA, and while the tyrosine kinase inhibitor imatinib suppresses FIP1L1‐PDGFRA‐dependent cell growth, ginkgolic acid or siRNA of PIAS1 has a synergistic effect with imatinib. In conclusion, our results suggest that sumoylation by PIAS1 is a potential target in the treatment of FIP1L1‐PDGFRA‐positive chronic eosinophilic leukemia.