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NISHIMURA NoriyukiGraduate School of Health Sciences / Faculty of Health SciencesProfessor
Researcher basic information
■ Research news- 06 Feb. 2020, New method for monitoring residual disease after treatment in children with neuroblastoma
- Pediatric cancer
- Neuroblastoma
- Leukemia
- Minimal Residual Disease
- Umbilical cord
- Mesenchymal Stem Cell
- Chronic Lung Disease
- Autism Spectrum Disorder
Research activity information
■ Paper- BACKGROUND: Bleomycin (BLM)-induced lung injury is characterized by mixed histopathologic changes with inflammation and fibrosis, such as observed in human patients with bronchopulmonary dysplasia, idiopathic pulmonary fibrosis, and chronic obstructive pulmonary disease. Although no curative therapies for these lung diseases exist, stem cell therapy has emerged as a potential therapeutic option. Multilineage-differentiating stress-enduring (Muse) cells are endogenous pluripotent- and macrophage-like stem cells distributed in various adult and fetal tissues as stage-specific embryonic antigen-3-positive cells. They selectively home to damaged tissue by sensing sphingosine-1-phosphate and replace the damaged/apoptotic cells by in vivo differentiation. Clinical trials for some human diseases suggest the safety and therapeutic efficacy of intravenously injected human leukocyte antigen-mismatched allogenic Muse cells from adult bone marrow (BM) without immunosuppressant. Here, we evaluated the therapeutic effects of human Muse cells from preterm and term umbilical cord (UC), and adult BM in a rat BLM-induced lung injury model. METHODS: Rats were endotracheally administered BLM to induce lung injury on day 0. On day 3, human preterm UC-Muse, term UC-Muse, or adult BM-Muse cells were administered intravenously without immunosuppressants, and rats were subjected to histopathologic analysis on day 21. Body weight, serum surfactant protein D (SP-D) levels, and oxygen saturation (SpO2) were monitored. Histopathologic lung injury scoring by the Ashcroft and modified American Thoracic Society document scales, quantitative characterization of engrafted Muse cells, RNA sequencing analysis, and in vitro migration assay of infused Muse cells were performed. RESULTS: Rats administered preterm- and term-UC-Muse cells exhibited a significantly better recovery based on weight loss, serum SP-D levels, SpO2, and histopathologic lung injury scores, and a significantly higher rate of both Muse cell homing to the lung and alveolar marker expression (podoplanin and prosurfactant protein-C) than rats administered BM-Muse cells. Rats receiving preterm-UC-Muse cells showed statistically superior results to those receiving term-UC-Muse cells in many of the measures. These findings are thought to be due to higher expression of genes related to cell migration, lung differentiation, and cell adhesion. CONCLUSION: Preterm UC-Muse cells deliver more efficient therapeutic effects than term UC- and BM-Muse cells for treating BLM-induced lung injury in a rat model.May 2024, Stem cell research & therapy, 15(1) (1), 147 - 147, English, International magazineScientific journal
- The dystrophin (DMD) gene is recognized for its significance in Duchenne muscular dystrophy (DMD), a lethal and progressive skeletal muscle disease. Some DMD patients, as well as model mice with muscular dystrophy (mdx), spontaneously develop various types of tumors, among which rhabdomyosarcoma (RMS) is the most prominent. By contrast, spindle cell sarcoma (SCS) has rarely been reported in patients or mdx mice. In this study, we aimed to use metabolomics to better understand the rarity of SCS development in mdx mice. Gas chromatography-mass spectrometry was employed to compare the metabolic profiles of spontaneously developed SCS and RMS tumors from mdx mice, and metabolite supplementation assays and silencing experiments were used to assess the effects of metabolic differences in SCS tumor-derived cells. The levels of 75 metabolites exhibited differences between RMS and SCS, 25 of which were significantly altered. Further characterization revealed downregulation of non-essential amino acids, including alanine, in SCS tumors. Alanine supplementation enhanced the growth, epithelial-mesenchymal transition, and invasion of SCS cells. Reduction of intracellular alanine via knockdown of the alanine transporter Slc1a5 reduced the growth of SCS cells. Lower metabolite secretion and reduced proliferation of SCS tumors may explain the lower detection rate of SCS in mdx mice. Targeting of alanine depletion pathways may have potential as a novel treatment strategy.Apr. 2024, American journal of physiology. Cell physiology, English, International magazineScientific journal
- High-risk neuroblastoma (HR-NB) patients remain far from obtaining optimal outcomes, with more than 50% relapse/regrowth rate despite current intensive multimodal therapy. This originated from the activation/proliferation of chemoresistant minimal residual disease (MRD). MRD with a significant prognostic was reported by several quantitative PCR (qPCR) or droplet digital PCR (ddPCR) assays quantitating different sets of NB-associated mRNAs (NB-mRNAs). The 7NB-mRNAs ddPCR assay quantitating CRMP1, DBH, DDC, GAP43, ISL1, PHOX2B, and TH mRNAs was reported to outperform other qPCR assays by a retrospective in-house observational study. In the present study, the Japan Children's Cancer Group (JCCG) Neuroblastoma Committee conducted a prospective multicenter observational study aimed at evaluating a prognostic value of MRD in bone marrow (BM-MRD) and peripheral blood (PB-MRD) detected by 7NB-mRNAs ddPCR assay. Between August 2018 and August 2022, 7 HR-NB patients who registered for JCCG clinical trials (JN-H-11 and JN-H-15) were enrolled. A total of 19 BM and 19 PB samples were collected, and 4/15 BM and 4/15 PB samples were classified as progressive disease (PD)/non-PD samples. BM-MRD and PB-MRD estimated area under curve (AUC) of 0.767 and 0.800 with a significant accuracy (AUC > 0.7). The present study validated a prognostic value of BM-MRD obtained by a previous study (AUC 0.723) and revealed the significant accuracy of PB-MRD as well as BM-MRD.Oct. 2023, Biology, 12(10) (10), English, International magazineScientific journal
- More than half of patients with high-risk neuroblastoma (HR-NB) experience relapse/regrowth due to the activation of chemoresistant minimal residual disease (MRD). MRD in patients with HR-NB can be evaluated by quantitating neuroblastoma-associated mRNAs (NB-mRNAs) in bone marrow (BM) and peripheral blood (PB) samples. Although several sets of NB-mRNAs have been shown to possess a prognostic value for MRD in BM samples (BM-MRD), MRD in PB samples (PB-MRD) is considered to be low and difficult to evaluate. The present report describes an HR-NB case presenting higher PB-MRD than BM-MRD before 1st and 2nd relapse/regrowth. A 3-year-old female presented with an abdominal mass, was diagnosed with HR-NB, and treated according to the nationwide standard protocol for HR-NB. Following systemic induction and consolidation therapy with local therapy, the patient achieved complete remission but experienced a 1st relapse/regrowth 6 months after maintenance therapy. The patient partially responded to salvage chemotherapy and anti-GD2 immunotherapy but had a 2nd relapse/regrowth 14 months after the 1st relapse/regrowth. Consecutive PB-MRD and BM-MRD monitoring revealed that PB-MRD was lower than BM-MRD at diagnosis (100 times) and 1st and 2nd relapse/regrowth (1,000 and 3 times) but became higher than BM-MRD before 1st and 2nd relapse/regrowth. The present case highlights that PB-MRD can become higher than BM-MRD before relapse/regrowth of patients with HR-NB.Sep. 2023, Oncology letters, 26(3) (3), 369 - 369, English, International magazine
- More than half of high-risk neuroblastoma (NB) patients have experienced relapse due to the activation of chemoresistant minimal residual disease (MRD) even though they are treated by high-dose chemotherapy with autologous peripheral blood stem cell (PBSC) transplantation. Although MRD in high-risk NB patients can be evaluated by quantitative PCR with several sets of neuroblastoma-associated mRNAs (NB-mRNAs), the prognostic significance of MRD in PBSC grafts (PBSC-MRD) is unclear. In the present study, we collected 20 PBSC grafts from 20 high-risk NB patients and evaluated PBSC-MRD detected by droplet digital PCR (ddPCR) with 7NB-mRNAs (CRMP1, DBH, DDC, GAP43, ISL1, PHOX2B, and TH mRNA). PBSC-MRD in 11 relapsed patients was significantly higher than that in 9 non-relapsed patients. Patients with a higher PBSC-MRD had a lower 3-year event-free survival (P = 0.0148). The present study suggests that PBSC-MRD detected by ddPCR with 7NB-mRNAs has a prognostic impact on high-risk NB patients.Oct. 2022, Heliyon, 8(10) (10), e10978, English, International magazineScientific journal
- (公社)日本小児科学会, Mar. 2022, 日本小児科学会雑誌, 126(3) (3), 552 - 552, Japanese神戸市東部療育センター診療所の開所後3年間のまとめ
- Neonatal jaundice has been suggested as a perinatal risk factor for autism spectrum disorder (ASD). We examined UGT1A1 polymorphisms to assess the potential of neonatal jaundice as a risk factor for ASD in children by using DNA extracted from preserved umbilical cord. In total, 79 children with ASD were genotyped for UGT1A1*28 (c.-41-40dup), UGT1A1*6 (c.211 G > A), and UGT1A1*27 (c.686 C > A). The allele frequency of UGT1A1*6 (OR = 1.34, p = 0.26) and UGT1A1*28 (OR = 0.80, p = 0.54) and the prevalence of UGT1A1*28/*6 diplotypes did not differ significantly from those in the control population. No UGT1A1*27 allele was detected in the subjects. ASD symptom assessment scores were not associated with UGT1A1*28/*6/*27 genotypes or UGT1A1*28/*6 diplotypes. These results suggest that neonatal jaundice is not significantly associated with ASD.Feb. 2022, Journal of autism and developmental disorders, 52(2) (2), 483 - 489, English, International magazineScientific journal
- Autoimmune enteropathy (AIE) is a rare disease, characterized by intractable diarrhea, villous atrophy of the small intestine, and the presence of circulating anti-enterocyte autoantibodies. Immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome, and mutations in FOXP3, which is a master gene of regulatory T cells (Tregs), are major causes of AIE. Recent studies have demonstrated that mutations in other Treg-associated genes, such as CD25 and CTLA4, show an IPEX-like phenotype. We present the case of a 13-year-old girl with CTLA4 haploinsufficiency, suffering from recurrent immune thrombocytopenic purpura and intractable diarrhea. We detected an autoantibody to the AIE-related 75 kDa antigen (AIE-75), a hallmark of the IPEX syndrome, in her serum. She responded well to a medium dose of prednisolone and a controlled dose of 6-mercaptopurine (6-MP), even after the cessation of prednisolone administration. Serum levels of the soluble interleukin-2 receptor and immunoglobulin G (IgG) were useful in monitoring disease activity during 6-MP therapy. In conclusion, autoimmune-mediated mechanisms, similar to the IPEX syndrome, may be involved in the development of enteropathy in CTLA4 haploinsufficiency. Treatment with 6-MP and monitoring of disease activity using serum levels of soluble interleukin-2 receptor and IgG is suggested for such cases.Jan. 2022, Intestinal research, 20(1) (1), 144 - 149, English, International magazine
- BACKGROUND: Blood transfusion is an important supportive care for high-risk neuroblastoma. When the number of transfusions increases, transfusion-associated adverse reactions may be more problematic. However, the factors determining the degree of myelosuppression and the number of transfusions during chemotherapy for high-risk neuroblastoma remain unclear. MATERIALS AND METHODS: We investigated patient factors determining the number of required transfusions in 15 high-risk neuroblastoma patients who received five courses of chemotherapy. Clinical data, cytokine profile and colony-forming assay with bone marrow samples at diagnosis were analysed. RESULTS: The required number of transfusions of both platelets and erythrocytes decreased once in the second course and then increased as the course progressed. The variability among cases increased as the chemotherapy course progressed. In cases of low peripheral blood platelet count and lower fibrinogen level at diagnosis, the number of platelet transfusions was higher during chemotherapy. In contrast, there was a negative correlation between the forming ability of granulocyte-macrophage or erythroid colonies and the number of erythrocyte transfusions in the latter period. CONCLUSION: In the early stages of chemotherapy, bone marrow infiltration in neuroblastoma and/or coagulopathy complication may cause thrombocytopenia and requirement of platelet transfusion; conversely, in the later stages, the number of erythrocyte transfusions may be defined by the patient's inherent hematopoietic ability. These factors may be useful in predicting the required number of transfusions.Jan. 2022, Vox sanguinis, 117(1) (1), 71 - 79, English, International magazineScientific journal
- Diamond-Blackfan anemia (DBA) is mainly caused by pathogenic variants in ribosomal proteins and 22 responsible genes have been identified to date. The most common causative gene of DBA is RPS19 [NM_001022.4]. Nearly 180 RPS19 variants have been reported, including three deep intronic variants outside the splicing consensus sequence (c.72-92A > G, c.356 + 18G > C, and c.411 + 6G > C). We also identified one case with a c.412-3C > G intronic variant. Without conducting transcript analysis, the pathogenicity of these variants is unknown. However, it is difficult to assess transcripts because of their fragility. In such cases, in vitro functional splicing assays can be used to assess pathogenicity. Here, we report functional splicing analysis results of four RPS19 deep intronic variants identified in our case and in previously reported cases. One splicing consensus variant (c.411 + 1G > A) was also examined as a positive control. Aberrant splicing with a 2-bp insertion between exons 5 and 6 was identified in the patient samples and minigene assay results also identified exon 6 skipping in our case. The exon 6 skipping transcript was confirmed by further evaluation using quantitative RT-PCR. Additionally, minigene assay analysis of three reported deep intronic variants revealed that none of them showed aberrant splicing and that these variants were not considered to be pathogenic. In conclusion, the minigene assay is a useful method for functional splicing analysis of inherited disease.Sep. 2021, Pediatric hematology and oncology, 38(6) (6), 515 - 527, English, International magazineScientific journal
- Neuroblastoma (NB) is the most common extracranial solid tumor in children and originates from sympathoadrenal or Schwann cell precursors derived from neural crest. These neural crest derivatives also constitute the hematopoietic and mesenchymal stem cells in bone marrow (BM) that is the most frequent site of NB metastasis and relapse. In NB patients, NB cells have been pathologically detected in BM and peripheral blood (PB), and minimal residual disease (MRD) in BM and PB (BM-MRD and PB-MRD) can be monitored by quantitating several sets of NB-associated mRNAs (NB-mRNAs). Although previous studies have shown varying degrees of correlation between BM-MRD and PB-MRD, the underlying factors and/or mechanisms remains unknown. In the present study, we determined the levels of BM-MRD and PB-MRD by quantitating seven NB-mRNAs in 133 pairs of concurrently collected BM and PB samples from 19 high-risk NB patients with clinical disease evaluation, and examined their correlation in overall and subgroups of sample pairs. The levels of BM-MRD and PB-MRD were moderately (r = 0.418, p < 0.001) correlated with each other in overall sample pairs. The correlation became strong (r = 0.725, p < 0.001), weak (r = 0.284, p = 0.008), and insignificant (p = 0.194) in progression, stable, and remission subgroups of sample pairs, respectively. It also became stronger in subgroups of sample pairs with poor treatment responses and poor prognostic factors. Present study suggests that MRD in high-risk NB shows a dynamic and disease burden-dependent correlation between BM and PB.Aug. 2021, Translational oncology, 14(8) (8), 101019 - 101019, English, International magazineScientific journal
- Vanillylmandelic acid (VMA), homovanillic acid (HVA), neuron-specific enolase (NSE) and lactate dehydrogenase (LDH) are classical tumor markers and are used as standard clinical evaluations for patients with neuroblastoma (NB). Minimal residual disease (MRD) can be monitored by quantifying several sets of NB-associated mRNAs in the bone marrow (BM) and peripheral blood (PB) of patients with NB. Although MRD in BM and PB has been revealed to be a strong prognostic factor that is independent of standard clinical evaluations, its interrelation with tumor markers remains uncharacterized. The present study determined the levels of tumor markers (VMA, HVA, NSE and LDH) and MRD (BM-MRD and PB-MRD) in 133 pairs of concurrently collected BM, PB and urine samples from 19 patients with high-risk NB. The patients were evaluated during the entire course of treatment, which included 10 diagnoses, 32 treatments, 36 post-treatment, 9 relapses and 46 post-relapse sample pairs. The level of BM-MRD and PB-MRD was determined by quantifying 7 NB-mRNAs (collapsin response mediator protein 1, dopamine beta-hydroxylase, dopa decarboxylase, growth-associated protein 43, ISL LIM homeobox 1, pairedlike homeobox 2b and tyrosine hydroxylase) using droplet digital PCR. In overall sample pairs, tumor markers (VMA, HVA, NSE and LDH) demonstrated weak but significant correlations (P<0.011) with BM-MRD and PB-MRD. In subgroups according to each patient evaluation, the degree of correlation between tumor markers and MRD became stronger in patients with adrenal gland tumors, BM metastasis at diagnosis and relapse/regrowth compared with overall sample pairs. In contrast, tumor markers demonstrated variable correlations with MRD in subgroups according to each sample evaluation (BM infiltration at sampling, collection time point and disease status). The results suggested that tumor markers may demonstrate limited correlation with MRD in patients with high-risk NB.Jul. 2021, Molecular and clinical oncology, 15(1) (1), 137 - 137, English, International magazineScientific journal
- Rapidly growing nontuberculous mycobacteria should be considered if GPRs gram-positive rods are detected in blood cultures 2-3 days after the blood sample collection.Feb. 2021, Clinical case reports, 9(2) (2), 835 - 840, English, International magazine
- WILEY, Dec. 2020, PEDIATRIC BLOOD & CANCER, 67, EnglishPrognostic Values of Tumor Markers and Minimal Residual Disease Detected by 7NB-mRNAs ddPCR Assay for High-Risk Neuroblastoma Patients
- We herein describe soft tissue tumor arising in the lower extremity of a pediatric patient. The tumor displayed a unique and wide range of histological features, sheet-like and cohesive growth pattern consisting of enlarged round to epithelioid atypical cells with a large alveolar and pseudopapillary histological architecture, focally mimicking alveolar soft part sarcoma and MiT family translocation renal cell carcinoma. Tumor cells were focally immunoreactive for cytokeratin, S-100, and EMA. RNA sequencing identified a novel in-frame NR1D1 (exon 5)-MAML1 (exon 2) gene rearrangement resulting in the formation of a putative chimeric protein containing the N-terminal C4-type zing finger domains of NR1D1 and the C-terminal MAML1 protein, which was confirmed by subsequent RT-PCR, Sanger sequencing, and FISH assay. To the best of our knowledge, NR1D1-MAML1 fusion has not yet been described in any neoplasms, suggesting the emergence of a novel tumor entity.Dec. 2020, Virchows Archiv : an international journal of pathology, 477(6) (6), 891 - 895, English, International magazine[Refereed]Scientific journal
- OBJECTIVE: To investigate the prevalence of seizures/febrile seizures in children up to 3 years of age and examine the effects of gestational age at birth on the risk for febrile seizures. DESIGN: Retrospective longitudinal population-based cohort study. SETTING: Kobe City public health center, Kobe, Japan, from 2010 to 2018. PARTICIPANTS: Children who underwent a medical check-up at 3 years of age. METHODS: Information regarding seizures was collected from the parents of 96 014 children. We identified the occurrence of seizure/febrile seizure in 74 017 children, whose gestational ages at birth were noted. We conducted a multivariate analysis with the parameter, gestational age at birth, to analyse the risk of seizure. We also stratified the samples by sex and birth weight (<2500 g or not) and compared the prevalence of seizure between those with the term and late preterm births. RESULTS: The prevalence of seizure was 12.1% (11.8%-12.3%), 13.2% (12.2%-14.4%), 14.6% (12.4%-17.7%) and 15.7% (10.5%-22.8%) in children born at 37-41, 34-36, 28-33 and 22-27 gestational weeks, respectively. The prevalence of febrile seizures was 9.0% (8.8%-9.2%), 10.5% (9.5%-11.5%), 11.8% (9.7%-14.5%) and 11.2% (6.9%-17.7%) in children born at 37-41, 34-36, 28-33 and 22-27 gestational weeks, respectively. Male was an independent risk factor for seizures (OR: 1.15, 95% CI 1.09 to 1.20; absolute risk increase 0.014, 95% CI 0.010 to 0.019) and febrile seizures (OR: 1.21, 95% CI 1.15 to 1.28; absolute risk increase 0.016, 95% CI 0.012 to 0.020), respectively. Late preterm birth was not associated with an increased risk of seizure/febrile seizure. CONCLUSIONS: Although very preterm birth may increase the risk of seizure/febrile seizure, the risk associated with late preterm birth is considerably small and less than that associated with male.Sep. 2020, BMJ open, 10(9) (9), e035977, English, International magazineScientific journal
- PURPOSE: To evaluate barbiturate anaesthetic therapy using thiamylal for febrile refractory status epilepticus (fRSE) in children. METHODS: This was a review of a prospectively-collected database between April 2012-March 2016 for fRSE cases treated with thiamylal anaesthetic therapy in a single paediatric hospital in Japan. The sample comprised 23 children (median age, 23 months) with fRSE that underwent thiamylal anaesthetic therapy for convulsive seizures lasting longer than 60 min, sustained after intravenous administration of benzodiazepine and non-benzodiazepine anticonvulsants. The intervention comprised protocol-based thiamylal anaesthetic therapy with bolus administration. We measured the dose and time required to achieve the burst suppression pattern (BSP) on electroencephalography, seizure recurrence, death, neurological sequelae, and complications. RESULTS: All patients except one reached the BSP. The thiamylal median dose until reaching the BSP was 27.5 mg/kg, and the median time from thiamylal administration to reaching the BSP was 109.5 min. There was one case of immediate treatment failure and one of withdrawal seizure, but no breakthrough seizure. No deaths occurred during treatment, and neurological sequelae occurred in four cases (17%). Vasopressors were administered in all cases. Other complications included 11 cases of pneumonia and one of enterocolitis. CONCLUSION: We revealed the time and dose required to reach the BSP with thiamylal anaesthetic therapy using bolus administration in children. Our results suggested that reaching the BSP with bolus administration requires markedly less time than without bolus administration, rarely causes seizure recurrence in paediatric fRSE, and causes haemodynamic dysfunction and infections as often as observed without bolus administration.Aug. 2020, Seizure, 80, 12 - 17, English, International magazine[Refereed]Scientific journal
- 産業開発機構映像情報メディカル編集部, May 2020, 映像情報medical : a monthly journal of medical imaging and information, 52(5) (5), 40 - 45, Japanese開発途上国における小児がん診療能力強化事業5年間のまとめ[Refereed]
- The outcomes of osteosarcoma with poor prognostic factors, such as poor responders, metastatic disease at diagnosis, and relapsed or refractory disease, are poor. We reviewed the clinical records of the patients diagnosed with osteosarcoma at our institute between 2004 and 2018 who received high-dose chemotherapy followed by autologous stem cell transplantation (ASCT) in our institute. Ten patients of osteosarcoma with poor responder, refractory status, and metastatic disease at diagnosis received high-dose chemotherapy followed by ASCT. Four patients underwent high-dose chemotherapy followed by ASCT with the conditioning regimen consisted of thiotepa and melphalan (MEL). Five patients underwent high-dose chemotherapy followed by ASCT with the conditioning regimen consisted of intravenous busulfan (BU) and MEL. One patient underwent tandem high-dose chemotherapy followed by ASCT with BU and MEL followed by carboplatin and etoposide. None of the ten patients died of regimen related toxicities. None of the five patients with poor responders who underwent high-dose chemotherapy followed by ASCT as part of consolidation therapy died of disease after ASCT. High-dose chemotherapy followed by ASCT might be effective for poor responders in osteosarcoma.May 2020, Pediatric hematology and oncology, 37(4) (4), 337 - 343, English, International magazine[Refereed]Scientific journal
- (公社)日本小児科学会, Feb. 2020, 日本小児科学会雑誌, 124(2) (2), 293 - 293, Japanese当院における小児がん患者の妊孕性温存について
- (公社)日本小児科学会, Feb. 2020, 日本小児科学会雑誌, 124(2) (2), 271 - 271, JapaneseNICU退院児における感覚特性,発達障害特性の検討
- Monitoring of several sets of neuroblastoma-associated mRNAs (NB-mRNAs) by real-time quantitative PCR (qPCR) can be used to evaluate minimal residual disease in NB patients. Droplet digital PCR (ddPCR) is an adaption of qPCR that potentially provides simpler and more reproducible detection of low levels of mRNAs. However, whether minimal residual disease in NB patients can be monitored by ddPCR using a set of NB-mRNAs is not yet tested. In this study, 208 bone marrow (BM) and 67 peripheral blood samples were retrospectively collected from 20 high-risk NB patients with clinical disease evaluation at two Japanese centers between 2011 and 2018, and level of each NB-mRNA (CRMP1, DBH, DDC, GAP43, ISL1, PHOX2B, and TH mRNAs) was determined by ddPCR. Level of 7NB-mRNAs (defined as the combined signature of each NB-mRNA) was higher in BM than peripheral blood, but correlated significantly with each other. In accordance with disease burden, it varied with disease status (remission, stable, or progression) and collection time point (diagnosis, treatment, post-treatment, or relapse). In 73 post-treatment BM samples, it was significantly higher in 17 relapsed/regrown samples than in 56 nonrelapsed/nonregrown samples. Furthermore, ddPCR had a better prognostic value than qPCR in detecting 7NB-mRNAs in the same 73 post-treatment BM samples. This study suggests that ddPCR detection of 7NB-mRNAs is significantly associated with tumor relapse/regrowth in high-risk NB patients.Feb. 2020, The Journal of molecular diagnostics : JMD, 22(2) (2), 236 - 246, English, International magazine[Refereed]Scientific journal
- The present case underscores the importance of considering the association of severe thrombocytopenia or immune thrombocytopenia with cytomegalovirus (CMV) infection because CMV-induced thrombocytopenia occasionally requires antiviral therapy.Jan. 2020, Clinical case reports, 8(1) (1), 75 - 78, English, International magazine[Refereed]
- Dec. 2019, PEDIATRIC BLOOD & CANCER, 66, S117 - S118Spontaneously Developed Undifferentiated Sarcoma and Rhabdomyosarcoma in mdx Mice[Refereed]
- 日本癌学会, Sep. 2019, 日本癌学会総会記事, 78回, E - 1060, English神経芽腫の七つのmRNA発現に関する骨髄と末梢血サンプルの相関(Expression of seven neuroblastoma-associated mRNAs is correlated between bone marrow and peripheral blood samples)[Refereed]
- OBJECTIVE: Although the mortality among previously healthy children with acute encephalopathy (AE) is approximately 5%, their detailed clinical course has not been clarified. The objective of the present study was to describe the detailed clinical course, in minutes, of fatal AE. METHODS: We retrospectively reviewed the medical records of five patients (from 6 months to 14 years of age) who previously had no neurological disorders and were diagnosed with brain death due to AE between 2002 and 2018 at Kobe Children's Hospital. RESULTS: The initial clinical symptoms were convulsion in three cases and impaired consciousness in two. The earliest noted brain imaging abnormality was 7.5 h after neurological symptom detection. Liver enzymes and creatinine levels increased at initial examination, and sodium elevated gradually. All patients met the criteria of systemic inflammatory response syndrome, disseminated intravascular coagulation, and shock within 14 h of symptom detection. High dose steroids and targeted temperature management were initiated 3.5-14 h after onset. Despite these therapies, patients were diagnosed with brain death from 16 h to 4 days after initial neurological symptoms. AE diagnoses were made between 4 h 29 min and 4 days after initial neurological symptoms and included hemorrhagic shock and encephalopathy syndromes, Reye-like syndrome, and acute necrotizing encephalopathy in two, two, and one patient(s), respectively. CONCLUSIONS: We revealed the time series' of clinical events (e.g. SIRS, shock, DIC, AE diagnosis, brain death, and treatments) and laboratory findings relative to initial neurological symptom in fatal AE.Sep. 2019, Brain & development, 41(8) (8), 691 - 698, English, International magazine[Refereed]Scientific journal
- BACKGROUND: Seizures and/or impaired consciousness accompanied by fever without known etiology (SICF) is common in the pediatric emergency setting. No optimal strategy for the management of SICF in childhood currently exists. We previously demonstrated the effectiveness of targeted temperature management (TTM) against SICF with a high risk of morbidity; however, some patients with SICF develop neurological sequelae despite TTM, which necessitate additional neuroprotective treatment. The clinical characteristics of these severe cases have not been studied. Accordingly, the aim of this study was to identify the clinical characteristics of children with SICF who exhibit poor outcomes after TTM. METHODS: The medical records of children admitted to Kobe Children's Hospital (Kobe, Japan) between October 2002 and September 2016 were retrospectively reviewed. Patients with SICF treated using TTM were included and divided into the satisfactory and poor outcome groups. Univariate and multivariate logistic regression analyses were used to compare clinical characteristics and laboratory findings between the two groups. RESULTS: Of the 73 included children, 10 exhibited poor outcomes. Univariate logistic regression analysis revealed that acute circulatory failure before TTM initiation, the use of four or more types of anticonvulsants, methylprednisolone pulse therapy, and an aspartate aminotransferase (AST) level ≥73 IU/L were associated with poor outcomes. Multivariate logistic regression analysis identified an elevated AST level as a significant independent predictor of a poor outcome. CONCLUSIONS: An elevated AST level within 12 h of onset in children with SICF is an independent predictor of a poor outcome after TTM initiated within 24 h of onset.Aug. 2019, Brain & development, 41(7) (7), 604 - 613, English, International magazine[Refereed]Scientific journal
- (一社)日本小児精神神経学会, Jun. 2019, 日本小児精神神経学会プログラム・抄録集, 121回, 43 - 43, Japanese修正18〜24ヵ月のNICU退院児における感覚特性の検討[Refereed]
- The detailed clinical time course in acute disseminated encephalomyelitis (ADEM) from initial symptoms, through exacerbation, to remission has not been widely reported. Hence, this study aimed to investigate the clinical time course of pediatric ADEM. This was a multicenter retrospective study based on registry data from medical chart reviews. The study included children who met the international consensus diagnostic criteria for ADEM. The patients comprised 18 boys and 6 girls, with a mean age of 5.5 ± 3.3 years at onset. From onset, the time until peak neurological symptoms, time until initial improvement, and time until full recovery was 3.1 ± 3.7 days, 6.0 ± 4.5 days, and 26 ± 34 days, respectively. Twenty-three (96%) patients were treated with high-dose methylprednisolone (mPSL) with a mean duration of 4.1 ± 4.0 days from onset. The condition of 15 patients (65%) improved within 3 days of high-dose mPSL initiation, whereas, that of four patients began to improve after >5 days of high-dose mPSL initiation. Only one patient (4%) did not achieve full recovery despite treatment with high-dose mPSL, intravenous immunoglobulin, and plasma exchange. This study presents the detailed clinical time course in pediatric ADEM in Japan. Progression of neurologic deficits typically lasts a few days, with initial improvement in 1 week leading to full recovery within 1 month.Jun. 2019, Brain & development, 41(6) (6), 531 - 537, English, International magazine[Refereed]Scientific journal
- 産業開発機構映像情報メディカル編集部, May 2019, 映像情報medical : a monthly journal of medical imaging and information, 51(5) (5), 62 - 65, Japaneseインドネシアにおける小児がん診療の現状と課題[Refereed]
- 産業開発機構映像情報メディカル編集部, Apr. 2019, 映像情報medical : a monthly journal of medical imaging and information, 51(4) (4), 42 - 48, Japanese開発途上国における小児がんの診療能力強化 : 国際的に注目されつつある課題である小児がんへの対応に関する研修[Refereed]
- OBJECTIVES: To evaluate the proportion of children presenting to the emergency department (ED) with altered mental status who demonstrate nonconvulsive seizures on reduced-lead electroencephalography (EEG), and to further investigate the characteristics, treatment, and outcomes in these patients compared with patients without nonconvulsive seizures. STUDY DESIGN: In this retrospective cohort study, we reviewed the database and medical records of pediatric patients (aged <18 years) in a single ED between May 1, 2016, and April 30, 2018. We first determined the proportion of nonconvulsive seizures among patients with altered mental status (Glasgow Coma Scale <15). We then compared the clinical presentation, demographic data, clinical diagnosis, EEG results, treatment, and outcomes of patients with altered mental status with nonconvulsive seizures and those without nonconvulsive seizures. RESULTS: In total, 16.9% of the patients with altered mental status (41 of 242; 95% CI, 12.2%-21.6%) evaluated by EEG had detectable nonconvulsive seizure, equivalent to 4.4% (41 of 932) of all patients with altered mental status presenting at our hospital. More than 80% of patients monitored for nonconvulsive seizures had a previous history of seizures, often febrile. Patients with nonconvulsive seizures were older (median, 68.5 vs 36.1 months) and had a higher Pediatric Cerebral Performance Category score at presentation (median, 2.0 vs 1.0). In addition, the proportion of patients admitted to the intensive care unit was significantly higher in the patients with nonconvulsive seizures (30.3% vs 15.0%). However, total duration of hospitalization, neurologic sequelae, and 30-day mortality rate did not differ between the 2 groups. CONCLUSIONS: A relatively high percentage of pediatric patients with altered mental status in the ED experience nonconvulsive seizures. The use of reduced-lead EEG monitoring in the ED might facilitate the recognition and treatment of nonconvulsive seizures, especially among patients with a history of seizures.Apr. 2019, The Journal of pediatrics, 207, 213 - 219, English, International magazine[Refereed]Scientific journal
- Mesenchymal stem cells (MSCs) have considerable therapeutic potential and attract increasing interest in the biomedical field. MSCs are originally isolated and characterized from bone marrow (BM), then acquired from tissues including adipose tissue, synovium, skin, dental pulp, and fetal appendages such as placenta, umbilical cord blood (UCB), and umbilical cord (UC). MSCs are a heterogeneous cell population with the capacity for (1) adherence to plastic in standard culture conditions, (2) surface marker expression of CD73+/CD90+/CD105+/CD45-/CD34-/CD14-/CD19-/HLA-DR- phenotypes, and (3) trilineage differentiation into adipocytes, osteocytes, and chondrocytes, as currently defined by the International Society for Cellular Therapy (ISCT). Although BM is the most widely used source of MSCs, the invasive nature of BM aspiration ethically limits its accessibility. Proliferation and differentiation capacity of MSCs obtained from BM generally decline with the age of the donor. In contrast, fetal MSCs obtained from UC have advantages such as vigorous proliferation and differentiation capacity. There is no ethical concern for UC sampling, as it is typically regarded as medical waste. Human UC starts to develop with continuing growth of the amniotic cavity at 4-8 weeks of gestation and keeps growing until reaching 50-60 cm in length, and it can be isolated during the whole newborn delivery period. To gain insight into the pathophysiology of intractable diseases, we have used UC-derived MSCs (UC-MSCs) from infants delivered at various gestational ages. In this protocol, we describe the isolation and characterization of UC-MSCs from fetuses/infants at 19-40 weeks of gestation.Jan. 2019, Journal of visualized experiments : JoVE, 143(143) (143), e58806, English, International magazine[Refereed]Scientific journal
- Neuroblastoma is a common extracranial solid tumor of neural crest (NC) origin that accounts for up to 15% of all pediatric cancer deaths. The disease arises from a transient population of NC cells that undergo an epithelial-mesenchymal transition (EMT) and generate diverse cell-types and tissues. Patients with neuroblastoma are characterized by their extreme heterogeneity ranging from spontaneous regression to malignant progression. More than half of newly diagnosed patients present highly metastatic tumors and are stratified into a high-risk group with dismal outcome. As many as 20% of high-risk patients have residual disease that is refractory or progressive during induction chemotherapy. Although a majority of high-risk patients achieve remission, larger part of those patients has minimal residual disease (MRD) that causes relapse even after additional consolidation therapy. MRD is composed of drug-resistant tumor cells and dynamically presented as cancer stem cells (CSCs) in residual tumors, circulating tumor cells (CTCs) in peripheral blood (PB), and disseminated tumor cells (DTCs) in bone marrow (BM) and other metastatic sites. EMT appears to be a key mechanism for cancer cells to acquire MRD phenotypes and malignant aggressiveness. Due to the restricted availability of residual tumors, PB and BM have been used to isolate and analyze CTCs and DTCs to evaluate MRD in cancer patients. In addition, recent technical advances make it possible to use circulating tumor DNA (ctDNA) shed from tumor cells into PB for MRD evaluation. Because MRD can be detected by tumor-specific antigens, genetic or epigenetic changes, and mRNAs, numerous assays using different methods and samples have been reported to detect MRD in cancer patients. In contrast to the tumor-specific gene-rearrangement-positive acute lymphoblastic leukemia (ALL) and the oncogenic fusion-gene-positive chronic myelogenous leukemia (CML) and several solid tumors, the clinical significance of MRD remains to be established in neuroblastoma. Given the extreme heterogeneity of neuroblastoma, dynamics of MRD in neuroblastoma patients will hold a key to the clinical validation. In this review, we summarize the biology and detection methods of cancer MRD in general and evaluate the available assays and clinical significance of neuroblastoma MRD to clarify its dynamics in neuroblastoma patients.2019, Frontiers in oncology, 9, 455 - 455, English, International magazine[Refereed]Scientific journal
- CLOVES syndrome is characterized by congenital lipomatous overgrowth, vascular malformation, epidermal nevi, and scoliosis/spinal malformation. It is caused by somatic mosaicism of gain-of-function variants of PIK3CA. Here, we describe a novel case of a 5-year-old Japanese girl with CLOVES and concurrent pancreatic steatosis. She had a recurrent somatic mutation in PIK3CA (NM_006218.3: c.1357G>A, p.Glu453Lys), elevated HbA1c levels, and pancreatic steatosis. This case indicates that pancreatic screening is critical for PIK3CA-related disorders.2019, Human genome variation, 6, 31 - 31, English, International magazine[Refereed]Scientific journal
- Although previous studies have investigated the influence of antiepileptic drugs (AEDs) on lipid profiles and thyroid hormone levels, there is little evidence regarding the effects of levetiracetam (LEV). Therefore, we conducted a prospective longitudinal study to evaluate the effects of LEV and carbamazepine (CBZ) treatment on lipid profile and thyroid hormone levels in patients newly diagnosed with epilepsy. Inclusion criteria were as follows: (a) age between 4 and 15 years, (b) diagnosis of epilepsy with at least two focal seizures within a year, and (c) newly treated with LEV or CBZ monotherapy. Serum lipid profile and thyroid hormone levels were measured before and after 1 and 6 months of AED initiation. Among the 21 included patients (LEV: 13 patients, CBZ: 8 patients), all but one patient in the LEV group continued AED monotherapy during the study period. Although triglyceride (TG) levels tended to be increased in the CBZ group (baseline: 58.3 ± 22.0 mg/dl, 1 month: 63.8 ± 21.6 mg/dl, 6 months: 92.3 ± 63.6 mg/dl, p = 0.22, analyses of variance (ANOVA)), there were no significant changes in total cholesterol (TC), TG levels, high-density lipoprotein cholesterol (HDL-C), or low-density lipoprotein cholesterol (LDL-C) in either group. Serum free thyroxine (fT4) levels were significantly decreased in the CBZ group (baseline: 1.15 ± 0.06 ng/dl, 1 month: 1.00 ± 0.16 ng/dl, 6 months: 0.98 ± 0.14 ng/dl, p = 0.03, ANOVA). In contrast, there were no significant changes in fT4 or thyroid-stimulating hormone (TSH) levels in the LEV group. The results of the present study suggest that LEV monotherapy does not affect lipid profile or thyroid function while CBZ monotherapy may cause thyroid dysfunction.Jan. 2019, Epilepsy & behavior : E&B, 90, 15 - 19, English, International magazine[Refereed]Scientific journal
- Dec. 2018, Pediatric blood & cancer, 65(12) (12), e27414, English, International magazine[Refereed]Scientific journal
- Nov. 2018, PEDIATRIC BLOOD & CANCER, 65, S95Spontaneous Development of Spindle Cell Sarcoma in mdx Mice[Refereed]
- OBJECTIVE: Asymmetric ventriculomegaly is often evident on brain magnetic resonance imaging (MRI) in very low birth weight infants (VLBWI) and is interpreted as white matter injury. However, no evaluation index for asymmetric left-right and anterior-posterior ventricular sizes has been established. METHODS: In this retrospective multicenter cohort study, brain T2-weighted MRI was performed at term-equivalent ages in 294 VLBWI born between 2009 and 2011. The value of a lateral ventricular index (LVI) to evaluate asymmetric ventricular size, as well as the relationship between the LVI value and walking at a corrected age of 18 months was investigated. At the level of the foramen of Monro in a horizontal slice, asymmetry between the left and right sides and between the anterior and posterior horns was identified by the corrected width and was detected by a low concordance rate and κ statistic value. An LVI representing the sum of the widths of the four horns of the lateral ventricle corrected for cerebral diameter was devised. RESULTS: Asymmetric left-right and anterior-posterior ventricular sizes were confirmed. The LVI value was significantly higher in the non-walking VLBWI group (n = 39) than in the walking VLBWI group (n = 255; 18.2 vs. 15.8, p = 0.02). An LVI cut-off value of 21.5 was associated with non-walking. Multivariate analysis revealed that an LVI value >21.5 was an independent predictor of walking disability at the corrected age of 18 months (odds ratio 2.56, p = 0.008). CONCLUSIONS: The LVI value calculated via MRI may predict walking disability at a corrected age of 18 months in VLBWI.Oct. 2018, Brain & development, 40(9) (9), 753 - 759, English, International magazine[Refereed]Scientific journal
- Characterization of Sensory Processing Function in Infants/Toddlers with Developmental Disorder自閉スペクトラム症(ASD)と知的能力障害(ID)はしばしば併存するが、両者の有無による感覚特性の差異に関する検討は乏しい。本研究では、発達の遅れや偏りを主訴に療育施設を受診した18〜36ヵ月の児を対象に、新版K式発達検査2001、小児自閉症評定尺度(CARS)を用いてID群(発達指数<70)、ASD群(CARS≧30点)を定義し、日本版乳幼児感覚プロファイル(ITSP-J)を用いて感覚処理機能を比較検討した。対象は60例で、ASD群/non ASD群:22例/38例、ID群/non ID群:29例/31例であった。ASD群は、non ASD群との間に、感覚処理機能7項目(4象限の4項目と5セクションの3項目)で有意差を認めたが、ID群とnon ID群間では有意差を認めた項目はなかった。発達の遅れや偏りを疑われた乳幼児の感覚特性は、IDの有無よりもASD特性の有無により強く関連した。(著者抄録)Last, 日本小児精神神経学会, Oct. 2018, 小児の精神と神経, 58(3) (3), 201 - 207, Japanese[Refereed]
- BACKGROUND: Children who present with seizure and/or impaired consciousness accompanied by fever without known etiology (SICF) may be diagnosed with either acute encephalopathy (AE) or febrile seizure (FS). Although approximately 5% of AE cases are fatal, it is difficult to identify fatal cases among children with SICF, which are often critical by the time of diagnosis. Thus, early prediction of outcomes for children with SICF, prior to diagnosis, may help to reduce mortality associated with AE. The aim of the present study was to identify clinical and laboratory risk factors for mortality acquired within 6 h of onset among children with SICF. METHODS: We retrospectively reviewed the medical records of children who had been admitted to Kobe Children's Hospital (Kobe, Japan) with SICF between October 2002 and September 2015. We compared clinical and laboratory characteristics acquired within 6 h of onset and outcomes between survivors and non-survivors using univariate and multivariate analyses. RESULTS: The survivor and non-survivor groups included 659 and nine patients, respectively. All patients in the non-survivor group received a final diagnosis of AE. Univariate analysis revealed significant differences between the groups with regard to seizure duration and the following laboratory parameters: aspartate transaminase (AST), alanine aminotransferase, lactate dehydrogenase, sodium, and lactate. The multivariate analysis identified AST as a significant independent factor associated with mortality. CONCLUSIONS: Elevation of AST within 6 h of onset is independently correlated with mortality in children with SICF. Our result may elucidate earlier intervention for patients with high risk of mortality.Aug. 2018, Brain & development, 40(7) (7), 552 - 557, English, International magazine[Refereed]Scientific journal
- BACKGROUND: CD134 (OX40), which is a cellular receptor for human herpesvirus-6B (HHV-6B) and expresses on activated T cells, may play a key role for HHV-6B replication after allogeneic hematopoietic stem cell transplantation (allo-HSCT). OBJECTIVES: Therefore, we examined the CD134 expression on T cells and HHV-6B replication after allo-HSCT, and analyzed the correlation between them. STUDY DESIGN: Twenty-three patients after allo-HSCT were enrolled. The percentages of CD134-positive cells within the CD4+ and CD8+ cell populations were measured by flow cytometry, and the viral copy number of HHV-6B was simultaneously quantified by real-time PCR. The correlation between CD134 and HHV-6B viral load was then statistically analyzed. RESULTS: HHV-6B reactivation occurred in 11 of 23 patients (47.8%). CD134 expression was seen on T cells and was coincident with the time of peak viral load. The percentage of CD134-positive cells decreased significantly when HHV-6B DNA disappeared (p = .005 in CD4+ T cells, p = .02 in CD8+ T cells). In the 4 patients who underwent umbilical cord blood transplantation (UCBT), the viral load varied with the percentage of CD134-positive cells. In the comparison between the HHV-6B reactivation group and non-reactivation group, maximum percentages of CD134-positive cells among CD4+ T cells in reactivation group were significantly higher than those in non-reactivation group (p = .04). CONCLUSIONS: This is the first study to show that a correlation of CD134 expression on T cells with HHV-6B replication after allo-HSCT, especially in UCBT. The results possibly indicate that CD134 on T cells plays a key role for HHV-6B replication after allo-HSCT.May 2018, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology, 102, 50 - 55, English, International magazine[Refereed]Scientific journal
- May 2018, J Clin Virol., 102, 50 - 55Expression of CD134 (OX40) on T cells is a trigger of human herpesvirus 6B reactivation and replication after hematopoietic cell transplantation.[Refereed]
- ETV6-ABL1 fusion is a rare but recurrent oncogenic lesion found in childhood B-cell precursor acute lymphoblastic leukemia (BCP-ALL), without an established chromosomal abnormality, and is associated with poor outcome. In ETV6-ABL1-positive cases, an in-frame fusion produced by a complex rearrangement results in constitutive chimeric tyrosine kinase activity. Monosomy 7 is also a rare and unfavorable chromosomal abnormality in childhood BCP-ALL. Here, we report a 14-year-old female BCP-ALL patient with ETV6-ABL1 fusion combined with monosomy 7. She was admitted to our hospital because of persistent fever. Bone marrow nuclear cell count on admission was 855,000/µL with 90.0% blastic cells of lymphoid morphology. Blasts were positive for CD10, CD19, CD20, CD34, cyCD79a, cyTdT, HLA-DR, and CD66c, had a karyotype of 45, XX, - 7 [18/20] and a split signal for ABL1 FISH probe (92.7%), and were sensitive to tyrosine kinase inhibitors, imatinib and dasatinib, in vitro. ETV6-ABL1 fusion transcript was identified by whole transcriptome sequencing and confirmed by RT-PCR. She was treated with the high-risk protocol based on ALL-BFM 95, achieved complete remission (CR) after induction chemotherapy, and maintained CR for 4 months. To our knowledge, this is the first report of ETV6-ABL1 fusion combined with monosomy 7 in childhood BCP-ALL.May 2018, International journal of hematology, 107(5) (5), 604 - 609, English, Domestic magazine[Refereed]Scientific journal
- Association of congenital cytomegalovirus (CMV) infection with autism spectral disorder (ASD) has been suggested since 1980s. Despite the observed association, its role as a risk factor for ASD remains to be defined. In the present review, we systematically evaluated the available evidence associating congenital CMV infection with ASD using PubMed, Web of Science, Cochrane Library, and Embase databases. Any studies on children with CMV infection and ASD were evaluated for eligibility and three observational studies were included in meta-analysis. Although a high prevalence of congenital CMV infection in ASD cases (OR 11.31, 95% CI 3.07-41.66) was indicated, too few events (0-2 events) in all included studies imposed serious limitations. There is urgent need for further studies to clarify this issue.May 2018, Journal of autism and developmental disorders, 48(5) (5), 1483 - 1491, English, International magazine[Refereed]Scientific journal
- Objectives To identify clinical features that predict Group A streptococcal (GAS) pharyngitis in a Japanese paediatric primary emergency medical centre. Methods The prevalence of GAS pharyngitis according to age and body temperature (BT) was calculated among 3098 paediatric patients with pharyngitis. The numbers of GAS-positive and -negative patients for each clinical parameter, and each point increase in the McIsaac score were compared and likelihood ratios (LRs) were calculated. Results The prevalence of GAS pharyngitis was extremely low in patients aged < 1 (1.2%) and 1 year (3.9%). The GAS-positive rate was significantly higher in patients with a BT < 38.0°C compared with ≥ 38.0°C (30.0% vs. 19.8%). A BT ≥ 38.0°C was not a predictive finding for GAS pharyngitis (positive LR: 0.82). Rash was the most useful individual predictor, and a McIsaac score of 4 or 5 increased the probability; however, the positive LRs were 1.74 and 1.30, respectively. Conclusions The prevalence of GAS pharyngitis is extremely low in patients aged < 1 and 1 year, and a BT ≥ 38.0°C is not a predictive symptom. Although a rash and McIsaac score of 4 or 5 are associated with an increased probability, they cannot be used to confirm GAS infection.May 2018, The Journal of international medical research, 46(5) (5), 1791 - 1800, English, International magazine[Refereed]Scientific journal
- Several studies describing the diurnal occurrence of febrile seizures have reported greater seizure frequency early or late in the evening relative to midnight or early morning. However, no articles have reported on the diurnal occurrence of complex febrile seizure. Moreover, no studies have addressed the relationship between seizure severity and diurnal occurrence. We retrospectively evaluated complex febrile seizures in 462 children needing hospitalization, and investigated the relationship between severity and diurnal occurrence according to four categorized time periods (morning, afternoon, evening, and night). Our study showed that complex febrile seizures occurred most often in the evening, peaking around 18:00 (18:00-18:59), and least often at night (02:00-02:59). In addition, the frequency with which patients developed status epilepticus or needed anticonvulsant treatments was also lower during the night. However, the seizure duration and the proportion of the patients who needed anticonvulsant treatment were the same among the four time periods. Furthermore, we compared three subclasses (repeated episodes of convulsions, focal seizures, and prolonged seizures (≧15min)), two of the complex features (focal seizures and prolonged seizures), and all complex features among the four time periods. However, they were the same among the four time periods. Taken together, our data indicate that although the severity of seizures was stable over a 24-hour period, the occurrence of seizures in our cohort of pediatric patients with complex febrile seizures requiring hospitalization was highest in the evening and lowest at night.Mar. 2018, Epilepsy & behavior : E&B, 80, 280 - 284, English, International magazine[Refereed]Scientific journal
- (公社)日本小児科学会, Feb. 2018, 日本小児科学会雑誌, 122(2) (2), 233 - 233, Japanese5年以上のromiplostim投与後に無治療経過観察に至った幼児慢性免疫血小板減少性紫斑病[Refereed]
- We herein reported a 4-month-old boy with transplantation-associated atypical hemolytic uremic syndrome (TA-aHUS) who was successfully treated with eculizumab. The patient diagnosed with type 3 of familial hemophagocytic lymphohistiocytosis underwent cord blood transplantation. After transplantation, he developed TA-aHUS, but plasma exchanges were unsuccessful. We identified deletions in CFH-related gene 1 (del-CFHR1) by the multiplex ligation-dependent probe amplification testing procedure and CFH autoantibodies. Eculizumab has been administered to the patient, with a marked improvement being achieved in thrombocytopenia. He has been well except for the persistent microhematuria for a year after transplantation. Uncontrolled complement activation might be involved in the pathophysiology of TA-aHUS.Jan. 2018, Journal of pediatric hematology/oncology, 40(1) (1), e41-e44 - e44, English, International magazine[Refereed]Scientific journal
- Dec. 2017, Environmental Health and Preventive Medicine (Web), 22(1) (1), 22:15 (WEB ONLY), EnglishChanges in the numbers of patients with acute gastroenteritis after voluntary introduction of the rotavirus vaccine in a Japanese children's primary emergency medical center[Refereed]Scientific journal
- We report the case of a 10-year-old female with acute myeloid leukemia (AML) FAB M0 carrying a novel t(11;19)(q23;p13.1) MLL-ELL variant, in which intron 8 of MLL is fused to exon 6 of ELL. Complete remission, judged by morphology and cytogenetic analysis, was achieved after the conventional chemotherapy. Eight months after completion of therapy, the level of WT-1 in peripheral blood and the number of cells with the MLL-ELL fusion transcript resurged. However, the patient remained overtly healthy and the morphology in the bone-marrow smear was innocuous, with no sign of relapse or secondary leukemia. Without any evidence of relapse, the patient has been closely observed without any therapeutic intervention. For approximately 2 years after the completion of therapy, despite clonal proliferation of pre-leukemic cells with an MLL-ELL fusion gene, she has maintained complete remission. In this case, the rare variant form of MLL-ELL fusion that has been identified may be related to diminished leukemogenic capacity, resulting in the persistence of pre-leukemic status; an additional genetic abnormality may thus be necessary for full transformation of pre-leukemic cells.SPRINGER JAPAN KK, Dec. 2017, International journal of hematology, 106(6) (6), 847 - 851, English, Domestic magazine[Refereed]Scientific journal
- BackgroundNon-invasive transcutaneous bilirubin (TcB) monitoring has been widely used to screen for hyperbilirubinemia. TcB measured using the recently developed BiliCare system, however, has not been fully evaluated.MethodsOne hundred and seven TcB measurements were obtained from 82 Japanese newborns 35 weeks' gestational age within 2 weeks after birth. Measurements were taken at the scaphoid fossa, conchal cavity, and lobe of the ear using BiliCare. BiliCare TcB were compared with total serum bilirubin (TB) and TcB obtained using another bilirubinometer (JM-105).ResultsTranscutaneous bilirubin measured at all three sites significantly correlated with TB (r = 0.91, 0.93, and 0.93 at the scaphoid fossa, conchal cavity, and lobe, respectively). The mean differences were 0.1, -0.3, and 3.6 at the scaphoid fossa, conchal cavity, and lobe, respectively. BiliCare TcB at the scaphoid fossa significantly correlated with that using the JM-105 (r = 0.91). The mean difference was 0.0. BiliCare, however, produced a significantly higher and lower TcB than the JM-105 for TB <7 and 15 mg/dL, respectively.ConclusionsTranscutaneous bilirubin measurements taken at the scaphoid fossa or conchal cavity using BiliCare were more reliable than those at the earlobe. BiliCare TcB differed from those of the JM-105, for TB <7 or 15 mg/dL.WILEY, Oct. 2017, PEDIATRICS INTERNATIONAL, 59(10) (10), 1058 - 1063, English[Refereed]Scientific journal
- Oct. 2017, Pediatrics International, 59(10) (10), 1058‐1063, EnglishEvaluation of BiliCare transcutaneous bilirubin device in Japanese newborns[Refereed]Scientific journal
- (一社)日本周産期・新生児医学会, Jun. 2017, 日本周産期・新生児医学会雑誌, 53(2) (2), 476 - 476, Japanese
- (一社)日本小児神経学会, May 2017, 脳と発達, 49(Suppl.) (Suppl.), S360 - S360, Japanese
- (公社)日本小児科学会, May 2017, 日本小児科学会雑誌, 121(5) (5), 914 - 914, Japanese胸部CTにより速やかに診断し得たX連鎖性重症複合型免疫不全症の乳児[Refereed]
- A 10-year-old girl was referred to our hospital with left preauricular adenopathy and gingival swelling. She was diagnosed with B-cell precursor acute lymphoblastic leukemia (BCP-ALL) based on being positive for expressions of CD10, CD19, TdT and HLA-DR. She showed no CD20 expression at the time of diagnosis. Based on the initial diagnosis of BCP-ALL, induction chemotherapy for BCP-ALL was initiated. However, the blasts did not disappear from her peripheral blood. Bone marrow examination on day 33 identified 81.3% residual blasts with positive expressions of CD19, 20 and HLA-DR and negative CD10 and TdT expressions; these cells were morphologically and phenotypically different from those at the initial diagnosis. Based on cytogenetic studies, the final diagnosis was double-hit lymphoma/leukemia (DHL) with IgH-BCL2 and Igλ-MYC. Although dose intensive chemotherapy, including rituximab, led to complete remission, bone marrow and central nervous system relapse occurred. At relapse, blasts expressed CD10, CD19 and HLA-DR, but not CD20, findings the same as those at the onset. The patient died of the disease 44 days after cord blood transplantation with non-remission status. DHL in childhood is extremely rare and its prognosis is poor. The establishment of an effective treatment for DHL is highly anticipated.The Japanese Society of Hematology, Feb. 2017, 臨床血液, 58(2) (2), 143 - 149, Japanese[Refereed]
- Mesenchymal stem cells (MSCs) are a heterogeneous cell population that is isolated initially from the bone marrow (BM) and subsequently almost all tissues including umbilical cord (UC). UC-derived MSCs (UC-MSCs) have attracted an increasing attention as a source for cell therapy against various degenerative diseases due to their vigorous proliferation and differentiation. Although the cell proliferation and differentiation of BM-derived MSCs is known to decline with age, the functional difference between preterm and term UC-MSCs is poorly characterized. In the present study, we isolated UC-MSCs from 23 infants delivered at 22-40 weeks of gestation and analyzed their gene expression and cell proliferation. Microarray analysis revealed that global gene expression in preterm UC-MSCs was distinct from term UC-MSCs. WNT signaling impacts on a variety of tissue stem cell proliferation and differentiation, and its pathway genes were enriched in differentially expressed genes between preterm and term UC-MSCs. Cell proliferation of preterm UC-MSCs was significantly enhanced compared to term UC-MSCs and counteracted by WNT signaling inhibitor XAV939. Furthermore, WNT2B expression in UC-MSCs showed a significant negative correlation with gestational age (GA). These results suggest that WNT signaling is involved in the regulation of GA-dependent UC-MSC proliferation.HINDAWI LTD, 2017, Stem cells international, 2017, 8749751 - 8749751, English, International magazine[Refereed]Scientific journal
- Background: Acute gastroenteritis (AGE) is a major reason for presentation to pediatric primary emergency medical centers. Because rotavirus vaccines were introduced in November 2011 for voluntary vaccination in Japan, we analyzed the changes in the numbers of AGE patients. Methods: The number and proportion of patients visiting Kobe children's primary emergency medical center from January 2011 to February 2015 due to AGE, out of all visiting children, were investigated retrospectively. The rotavirus and norovirus epidemic periods were defined as the periods from March to June and from November to February, respectively, based on their disease prevalence. Results: In patients <= 2 years of age, the numbers and proportions of patients with AGE were significantly decreased from 2464/14098 (17%) in 2011 to 1888/12321 (15%) in 2014 (p < 0.01). In patients <= 2 and 3-5 years of age, significant decreases in AGE patients between 2011 and 2014 were observed during the rotavirus season (from 20% [1090/5329] to 14% [642/4482] in patients aged <= 2 years and from 23% [704/3047] to 20% [572/2807] in patients aged 3-5 years, p < 0.01 and p < 0.05, respectively), but not during the norovirus season (from 19% [834/4436] to 19% [797/4160] in patients aged <= 2 years and from 20% [679/3334] to 25% [710/2852] in patients aged 3-5 years). Conclusions: The estimated rotavirus vaccine coverage in our area increased from 1% in 2011 to 49% in 2014; this coverage may have resulted in a reduction in AGE patients, both directly and indirectly, in our Japanese children's primary emergency medical center.SPRINGER, 2017, ENVIRONMENTAL HEALTH AND PREVENTIVE MEDICINE, 22(1) (1), EnglishScientific journal
- BACKGROUND: Spinal muscular atrophy (SMA) is the most common genetic neurological disease leading to infant death. It is caused by loss of survival motor neuron (SMN) 1 gene and subsequent reduction of SMN protein in motor neurons. Because SMN is ubiquitously expressed and functionally linked to general RNA metabolism pathway, fibroblasts (FBs) are most widely used for the assessment of SMN expression in SMA patients but usually isolated from skin biopsy samples after the onset of overt symptoms. Although recent translational studies of SMN-targeted therapies have revealed the very limited time window for effective SMA therapies during perinatal period, the exact time point when SMN shortage became evident is unknown in human samples. In this study, we analyzed SMN mRNA and protein expression during perinatal period by using umbilical cord-derived mesenchymal stem cells (UC-MSCs) obtained from preterm and term infants. METHODS: UC-MSCs were isolated from 16 control infants delivered at 22-40 weeks of gestation and SMA fetus aborted at 19 weeks of gestation (UC-MSC-Control and UC-MSC-SMA). FBs were isolated from control volunteer and SMA patient (FB-Control and FB-SMA). SMN mRNA and protein expression in UC-MSCs and FBs was determined by RT-qPCR and Western blot. RESULTS: UC-MSC-Control and UC-MSC-SMA expressed the comparable level of MSC markers on their cell surface and were able to differentiate into adipocytes, osteocytes, and chondrocytes. At steady state, SMN mRNA and protein expression was decreased in UC-MSC-SMA compared to UC-MSC-Control, as observed in FB-SMA and FB-Control. In response to histone deacetylase inhibitor valproic acid, SMN mRNA and protein expression in UC-MSC-SMA and FB-SMA was increased. During perinatal development from 22 to 40 weeks of gestation, SMN mRNA and protein expression in UC-MSC-Control was positively correlated with gestational age. CONCLUSION: UC-MSCs isolated from 17 fetus/infant of 19-40 weeks of gestation are expressed functional SMN mRNA and protein. SMN mRNA and protein expression in UC-MSCs is increased with gestational age during perinatal development.FRONTIERS MEDIA SA, 2017, Frontiers in pediatrics, 5, 194 - 194, English, International magazine[Refereed]Scientific journal
- Gestational age (GA) is thought to affect height growth in small-for-gestational age (SGA) children. However, the GA-specific trajectories in body mass index (BMI) and early appearances of adiposity rebound (AR) have not been fully investigated in a cohort of Japanese SGA children. A longitudinal cohort study was conducted with 1063 SGA children born in Kobe, Japan, with sufficient records from birth to 3 years of age. Subjects were divided into subgroups based on GA: 39-41 weeks GA (n = 723), 37-38 weeks GA (n = 256), 34-36 weeks GA (n = 62), and <34 weeks GA (n = 22). Height and BMI were assessed at 4 months, 9 months, 1.5 years, and 3 years of age. The catch-up rate for height was GA-dependent. Most children with 39-41 weeks GA (91%) caught up by 4 months of age; however, lower GA was associated with a slower elevation in the catch-up rate. The BMI trajectory during the first 3 years was also GA-dependent, with a change in GA dependency at a boundary of 37 weeks GA. Approximately 7% of SGA children had already developed AR before 3 years of age. In conclusion, growth patterns during infancy and early childhood in SGA children differ depending on GA.NATURE PUBLISHING GROUP, Dec. 2016, Scientific reports, 6, 38659 - 38659, English, International magazine[Refereed]Scientific journal
- (一社)日本小児血液・がん学会, Nov. 2016, 日本小児血液・がん学会雑誌, 53(4) (4), 349 - 349, JapanesePET-MRI所見に基づく肺・下鼻甲介・腎生検が原疾患の診断に有効であった二次性血球貪食性リンパ組織球症の1例[Refereed]
- Neuroblastoma is the most common extracranial solid tumour in children and is histologically classified by its Schwannian stromal cells. Although having fewer Schwannian stromal cells is generally associated with more aggressive phenotypes, the exact roles of other stromal cells (mainly macrophages and fibroblasts) are unclear. Here, we examined 41 cases of neuroblastoma using immunohistochemistry for the tumour-associated macrophage (TAM) markers CD68, CD163, and CD204, and a cancer-associated fibroblast (CAF) marker, alpha smooth muscle actin (αSMA). Each case was assigned to low/high groups on the basis of the number of TAMs or three groups on the basis of the αSMA-staining area for CAFs. Both the number of TAMs and the area of CAFs were significantly correlated with clinical stage, MYCN amplification, bone marrow metastasis, histological classification, histological type, and risk classification. Furthermore, TAM settled in the vicinity of the CAF area, suggesting their close interaction within the tumour microenvironment. We next determined the effects of conditioned medium of a neuroblastoma cell line (NBCM) on bone marrow-derived mesenchymal stem cells (BM-MSCs) and peripheral blood mononuclear cell (PBMC)-derived macrophages in vitro. The TAM markers CD163 and CD204 were significantly up-regulated in PBMC-derived macrophages treated with NBCM. The expression of αSMA by BM-MSCs was increased in NBCM-treated cells. Co-culturing with CAF-like BM-MSCs did not enhance the invasive ability but supported the proliferation of tumour cells, whereas tumour cells co-cultured with TAM-like macrophages had the opposite effect. Intriguingly, TAM-like macrophages enhanced not only the invasive abilities of tumour cells and BM-MSCs but also the proliferation of BM-MSCs. CXCL2 secreted from TAM-like macrophages plays an important role in tumour invasiveness. Taken together, these results indicate that PBMC-derived macrophages and BM-MSCs are recruited to a tumour site and activated into TAMs and CAFs, respectively, followed by the formation of favourable environments for neuroblastoma progression. © 2016 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.WILEY-BLACKWELL, Oct. 2016, The Journal of pathology, 240(2) (2), 211 - 23, English, International magazine[Refereed]Scientific journal
- 兵庫県小児科医会, Oct. 2016, 兵庫県小児科医会報, 65(66) (66), 6 - 9, Japanese小児における幹細胞治療の最近の知見[Refereed]
- Neuroblastoma is an aggressive pediatric tumor accounting for ~15% of cancer-associated mortalities in children. Despite the current intensive therapy, >50% of high-risk patients experience tumor relapse or regrowth caused by the activation of minimal residual disease (MRD). Although several MRD detection protocols using various reverse transcription-quantitative polymerase chain reaction (RT-qPCR) markers have been reported to evaluate the therapeutic response and disease status of neuroblastoma patients, their clinical significance remains elusive. The present study reports two high-risk neuroblastoma patients, whose MRD was consecutively monitored using 11 RT-qPCR markers (CHRNA3, CRMP1, DBH, DCX, DDC, GABRB3, GAP43, ISL1, KIF1A, PHOX2B and TH) during their course of treatment. The two patients initially responded to the induction therapy and reached MRD-negative status. The patients' MRD subsequently became positive with no elevation of their urinary homovanillic acid, urinary vanillylmandelic acid and serum neuron-specific enolase levels at 13 or 19 weeks prior to the clinical diagnosis of tumor relapse or regrowth. The present cases highlight the possibility of consecutive MRD monitoring using 11 markers to enable an early detection of tumor relapse or regrowth in high-risk neuroblastoma patients.SPANDIDOS PUBL LTD, Aug. 2016, Oncology letters, 12(2) (2), 1119 - 1123, English, International magazine[Refereed]Scientific journal
- Increased serum levels of unconjugated bilirubin are associated with the development of brain damage in newborns. In current clinical settings, there are no methods for directly determining serum levels of unconjugated bilirubin. UnaG, a fluorescent protein from Japanese eel muscle that specifically binds to unconjugated bilirubin was used in this study. Linear regression analysis was carried out to compare unconjugated bilirubin levels measured by UnaG and conventional bilirubin oxidase methods. Unconjugated bilirubin levels in the serum of newborns who were untreated or treated with phototherapy were compared. Effects of interfering factors in the serum (conjugated bilirubin, hemoglobin, and lipid) on unconjugated bilirubin concentration measured by the UnaG method were also evaluated. Unconjugated bilirubin levels measured by the UnaG method were highly correlated with those determined by the bilirubin oxidase assay. Unconjugated bilirubin levels determined by bilirubin oxidase and UnaG assays were similar in serum samples containing conjugated bilirubin. The performance of the UnaG assay was unaffected by phototherapy and the presence of serum hemoglobin and lipid emulsion. These results demonstrate the clinical applicability of the UnaG method for direct measurement of unconjugated bilirubin levels in newborn serum.NATURE PUBLISHING GROUP, Jun. 2016, Scientific reports, 6, 28489 - 28489, English, International magazine[Refereed]Scientific journal
- Transient hyperphosphatasemia (TH) is defined as marked elevation of serum alkaline phosphatase (ALP), predominantly its bone or liver isoform. It is a rare condition and is usually detected on laboratory examination in patients without any clinical symptoms. In typical patients with TH, ALP spontaneously normalizes, but no apparent cause of TH has been identified. Some drugs are suspected triggers of TH, but no clear evidence of their association with TH has been shown to date. We herein report three cases of TH in pediatric patients. Two patients were treated with cyclosporine for frequently relapsing nephrotic syndrome, and one was also taking cyclosporine for aplastic anemia. Interestingly, ALP immediately decreased after termination of cyclosporine in two patients, whereas TH lasted 4 months in the one patient who continued cyclosporine. Clearly, cyclosporine is associated with the pathophysiology of TH in children.WILEY-BLACKWELL, May 2016, Pediatrics international : official journal of the Japan Pediatric Society, 58(5) (5), 429 - 30, English, International magazine[Refereed]Scientific journal
- (公社)日本小児科学会, Feb. 2016, 日本小児科学会雑誌, 120(2) (2), 316 - 316, Japanese神経芽腫患者の微小残存病変モニタリング[Refereed]
- (公社)日本小児科学会, Jan. 2016, 日本小児科学会雑誌, 120(1) (1), 84 - 84, Japanese左鎖骨部腫瘤を呈した生後6ヵ月の乳児例[Refereed]
- Neuroblastoma is an aggressive solid tumor that leads to tumor relapse in more than half of high-risk patients. Minimal residual disease (MRD) is primarily responsible for tumor relapses and may be detected in peripheral blood (PB) and bone marrow (BM) samples. To evaluate the disease status and treatment response, a number of MRD detection protocols based on either common or distinct markers for PB and BM samples have been reported. However, the correlation between the expression of MRD markers in PB and BM samples remains elusive in the clinical samples. In the present study, the expression of 11 previously validated MRD markers (CHRNA3, CRMP1, DBH, DCX, DDC, GABRB3, GAP43, ISL1, KIF1A, PHOX2B and TH) was determined in 23 pairs of PB and BM samples collected from seven high-risk neuroblastoma patients at the same time point, and the sample was scored as MRD-positive if one of the MRD markers exceeded the normal range. Although the number of MRD-positive samples was not significantly different between PB and BM samples, the two most sensitive markers for PB samples (CRMP1 and KIF1A) were different from those for BM samples (PHOX2B and DBH). There was no statistically significant correlation between the expression of MRD markers in the PB and BM samples. These results suggest that MRD markers were differentially expressed in PB and BM samples from high-risk neuroblastoma patients.SPANDIDOS PUBL LTD, Nov. 2015, Oncology letters, 10(5) (5), 3228 - 3232, English, International magazine[Refereed]Scientific journal
- Recent studies have reported that the absolute lymphocyte count (ALC) during induction therapy is predictive of treatment outcome in de novo acute lymphoblastic leukemia (ALL); however, the significance of ALC on outcomes remains controversial. In the present study, we assessed the significance of ALC at day 29 (ALC-29), the end of induction therapy, on outcomes in our Japanese cohort. The outcomes of 141 patients aged ≤18 years with newly diagnosed ALL who were enrolled on the JACLS ALL-02 at our hospitals were analyzed in terms of ALC-29. Patients with ALC-29 ≥750/μL (n = 81) had a superior 5-year EFS (95.2 ± 2.7 vs 84.3 ± 4.8 %, P = 0.016) and OS (100 vs 87.0 ± 4.7 %, P = 0.0062). A multivariate analysis identified ALC-29 ≥750/μL as a significant predictor of improved EFS and OS after controlling for confounding factors. A multiple linear regression model revealed a significant inverse relationship between the percentage of blasts in bone marrow on day 15 and ALC-29 (P = 0.005). These results indicate that ALC is a simple prognostic factor in childhood ALL, and, thus, has the potential to refine current risk algorithms.SPRINGER JAPAN KK, Nov. 2015, International journal of hematology, 102(5) (5), 594 - 601, English, Domestic magazine[Refereed]Scientific journal
- (一社)日本小児血液・がん学会, Oct. 2015, 日本小児血液・がん学会雑誌, 52(4) (4), 342 - 342, Japanese外来化学療法中(ALL B12臨床試験)の中心静脈カテーテルの管理についての検討[Refereed]
- 日本癌学会, Oct. 2015, 日本癌学会総会記事, 74回, J - 1346, English神経芽腫の発症・進展におけるRab6BとRab28の役割[Refereed]
- 日本癌学会, Oct. 2015, 日本癌学会総会記事, 74回, P - 3035, EnglishDENNドメイン蛋白質DENND2Aによる神経芽腫発の発症・進展の制御機構[Refereed]
- (一社)日本小児血液・がん学会, Oct. 2015, 日本小児血液・がん学会雑誌, 52(4) (4), 257 - 257, EnglishDENNドメイン蛋白質DENND2Aによる神経芽腫の制御機構(DENN domain protein DENND2A mediates the progression of neuroblastoma)[Refereed]
- (一社)日本小児血液・がん学会, Oct. 2015, 日本小児血液・がん学会雑誌, 52(4) (4), 257 - 257, English高リスク神経芽腫患者の骨髄と末梢血における微小残存病変の発現についての検討(Differential expression of minimal residual disease markers in peripheral blood and bone marrow samples from high-risk neuroblastoma patients)[Refereed]
- BACKGROUND: More than 90% of spinal muscular atrophy (SMA) patients show homozygous deletion of SMN1 (survival motor neuron 1). They retain SMN2, a highly homologous gene to SMN1, which may partially compensate for deletion of SMN1. Although the promoter sequences of these two genes are almost identical, a GCC insertion polymorphism has been identified at c.-320_-321 in the SMN1 promoter. We have also found this insertion polymorphism in an SMN2 promoter in an SMA patient (Patient A) who has SMA type 2/3. PURPOSE: The aims of this study were to determine the frequency of the GCC insertion polymorphism in SMA patients, and to evaluate its effect on SMN transcription efficiency. PATIENTS AND METHODS: Fifty-one SMA patients, including Patient A, were involved in this study. SMN2 transcript levels in white blood cells were measured by real-time polymerase chain reaction. Screening of the GCC insertion polymorphism was performed using denaturing high-pressure liquid chromatography. The transcription efficiency of the promoter with the insertion mutation was evaluated using a reporter-gene assay. RESULTS: All SMA patients in this study were homozygous for SMN1 deletion. Patient A retained two copies of SMN2, and showed only a small amount of SMN2 transcript in white blood cells. We detected a GCC insertion polymorphism at c.-320_-321 only in Patient A, and not in 50 other SMA patients. The polymorphism had a slight but significant negative effect on transcription efficiency. DISCUSSION AND CONCLUSION: Patient A was judged to be an exceptional case of SMA, because the GCC insertion polymorphism rarely exists in SMN1-deleted SMA patients. The GCC insertion polymorphism did not enhance the transcriptional efficiency of SMN2. Thus, this GCC insertion polymorphism in the SMN2 promoter may not be associated with the milder phenotype of the patient. Patient A suggests that there are other unknown factors modifying the clinical phenotype of SMA.ELSEVIER SCIENCE BV, Aug. 2015, Brain & development, 37(7) (7), 669 - 76, English, International magazine[Refereed]Scientific journal
- 兵庫県と県内4つの保健所設置市(神戸市・尼崎市・西宮市・姫路市)を対象とし、公衆衛生行政医師の「人数」「配置」「業務内容」などについてアンケート調査を行った。結果、兵庫県下の保健衛生行政関係部署には、県庁・市役所に7名、保健所に24名、神戸市の保健センターに9名、精神保健福祉センターに2名、県衛生研究所に1名の計43名の医師が存在し、4市の保健所にも複数の医師が配置されていた。県庁や市役所における医師業務は主として施策の企画立案や総合調整であり、保健所と保健センターでは感染症・結核業務や成人・母子保健等の事業に携わっていた。公衆衛生行政医師2名にインタビューを行い、「公衆衛生医師の道を選んだきっかけ」「これまでの経歴・仕事内容」「キャリアパスと人生設計」「公衆衛生医師を目指す医学部学生や若手研修医に勧める勉強法」などについて聴取したので、併せて報告した。(一社)神緑会, Aug. 2015, 神緑会学術誌, 31, 7 - 11, Japanese[Refereed]
- BACKGROUND: Spinal muscular atrophy is caused by survival motor neuron gene SMN1 mutations. SMN1 produces a full-length SMN1 protein isoform encoded by exons 1-7, and an axonal-SMN protein isoform encoded by exons 1-3 and intron 3. The axonal-SMN protein is expressed only in the embryonic period and plays a significant role in axonal growth. However, there has been no report on contribution of axonal-SMN to spinal muscular atrophy severity until now. PATIENTS: Two Japanese boys with spinal muscular atrophy type 1 in our study presented with generalized muscle weakness and respiratory insufficiency soon after birth and required an artificial ventilator from early infancy. Patient 1 was compound heterozygous for two SMN1 mutations, whole-gene deletion, and an intragenic mutation (c.819_820insT). He retained one copy of SMN1 producing the N-terminal part of SMN1 including axonal-SMN. On the other hand, patient 2 was homozygous for SMN1 deletion. Both of them showed the same copy number of spinal muscular atrophy-modifying genes, NAIP and SMN2. These findings suggested that the C-terminal domain of full-length SMN1 determined the severity, irrespective of presence or absence of axonal-SMN expression. CONCLUSION: In patient 1, the C-terminal domain of full-length SMN1 determined spinal muscular atrophy severity, rather than the axonal-SMN, one copy of which could be present and intact. The presence or absence of axonal-SMN may not impact disease severity in spinal muscular atrophy type 1 patients.ELSEVIER SCIENCE INC, Jun. 2015, Pediatric neurology, 52(6) (6), 638 - 41, English, International magazine[Refereed]Scientific journal
- Attention Deficit/Hyperactivity Disorder (ADHD): age related change of completion time and error rates of Stroop test.BACKGROUND: Attention Deficit/Hyperactivity Disorder (ADHD) is a common neurobehavioral problem in children throughout the world. The Stroop test has been widely used for the evaluation of ADHD symptoms. However, the age-related change of the Stroop test results has not been fully clarified until now. METHODS: Sixty-five ADHD and 70 age-matched control children aged 6-13 years were enrolled in this study. ADHD was diagnosed based on DSM-IV criteria. We examined the completion time and error rates of the Congruent Stroop test (CST) and Incongruent Stroop test (IST) in ADHD and control children. RESULTS: No significant difference was observed in the completion time for CST or IST between the ADHD and control children at 6-9 years old. However, ADHD children at 10-13 years old showed significantly delayed completion time for the CST and IST compared with controls of the same age. As for the error rates of the CST and IST, ADHD and control children at 6-9 years old showed no difference. However, error rates of CST and IST in the ADHD children at 10-13 years were significantly higher than those of control of the same age. CONCLUSIONS: Age may influence the results of Stroop test in ADHD children. For the ages of 10-13 years old, the Stroop test clearly separates ADHD children from control children, suggesting that it may be a useful screening tool for ADHD among preadolescent children.Apr. 2015, The Kobe journal of medical sciences, 61(1) (1), E19-26 - 26, English, Domestic magazine[Refereed]Scientific journal
- (一社)日本病理学会, Mar. 2015, 日本病理学会会誌, 104(1) (1), 273 - 273, Japanese神経芽腫の進展における腫瘍関連炎症細胞と腫瘍関連間質細胞の相互作用の解明[Refereed]
- Despite the introduction of 13-cis-retinoic acid (13-cis-RA) into the current chemotherapy, more than half of high-risk neuroblastoma patients have experienced tumor relapses driven by chemoresistant cancer stem cells (CSCs) that can be isolated by their ability to grow as spheres. Although aldehyde dehydrogenase (ALDH) has been used to characterize CSCs in certain cancers, ALDH remains elusive in neuroblastoma. In the present study, we determined ALDH activity and expression of its 19 isoforms in spheres and parental cells of neuroblastoma. ALDH activity and several ALDH isoforms were consistently induced in spheres of different neuroblastoma cells. While ALDH1A2, ALDH1L1 and ALDH3B2 expression was consistently induced in spheres and associated with the sphere and colony formation, only ALDH1A2 expression was significantly correlated with the poor prognosis of neuroblastoma patients. ALDH1A2 expression was further associated with the growth and undifferentiation of neuroblastoma xenografts and the resistance of neuroblastoma cells to 13-cis-RA. These results suggest that ALDH1A2 is involved in the regulation of CSC properties in neuroblastoma.SPANDIDOS PUBL LTD, Mar. 2015, International journal of oncology, 46(3) (3), 1089 - 98, English, International magazine[Refereed]Scientific journal
- (公社)日本小児科学会, Feb. 2015, 日本小児科学会雑誌, 119(2) (2), 410 - 410, Japanese化学療法中に血液培養陽性となった急性リンパ球白血病症例の臨床的検討[Refereed]
- SMA screening system using dried blood spots on filter paper: application of COP-PCR to the SMN1 deletion test.BACKGROUND: Spinal muscular atrophy (SMA) is a common neuromuscular disorder caused by mutations in SMN1. More than 95% of SMA patients carry homozygous SMN1 deletions. Thus, the SMN1 deletion test should be performed initially as part of the diagnostic process. However, SMN2, a highly homologous gene, hampers detection of SMN1 deletion. To differentiate between SMN1 and SMN2, many analysis methods have been developed yet they are not all available worldwide. AIM: To establish a simple but accurate SMN1-deletion detection system that can be used worldwide. METHODS: Fifty DNA samples (29 SMA patients and 21 controls) from dried blood spots (DBS) on filter paper were assayed. All participants had previously been screened for SMA by PCR-restriction fragment length polymorphism (PCR-RFLP) using DNA extracted from freshly collected blood. DNA was extracted from DBS that had been stored at room temperature (20-25℃) for between 1 and 8 years. Competitive oligonucleotide priming-PCR (COP-PCR) was performed to distinguish SMN1 and SMN2 exon7. RESULTS: DNA yield from an 11-mm diameter DBS circle was 21,171 ± 7,485 ng (mean ± SD), with an 260/280 OD ratio from 1.49 to 2.1(mean ± SD; 1.67 ±0.13). Nucleotide sequencing confirmed gene-specific amplification of SMN1 and SMN2 by COP-PCR. SMN1 and SMN2 COP-PCR results are completely consistent with those obtained by PCR-RFLP. CONCLUSION: We have combined DNA extraction from DBS on filter paper with COP-PCR that specifically detects SMN1 and SMN2, establishing a new SMN1-deletion detection system with practical application worldwide.Jan. 2015, The Kobe journal of medical sciences, 60(4) (4), E78-85 - 85, English, Domestic magazine[Refereed]Scientific journal
- 2015, 臨床細胞分子遺伝, 20, 6 - 8, Japanese神経芽腫の細胞分子遺伝学Scientific journal
- BACKGROUND: Spinal muscular atrophy (SMA) is a common neuromuscular disorder caused by mutation of the survival of the motor neuron 1 (SMN1) gene. More than 95% of SMA patients carry a homozygous deletion of SMN1. SMA can be screened for by polymerase chain reaction and high-resolution melting analysis (PCR-HRMA) using DNA extracted from dried blood spots (DBSs) stored on filter paper. However, there are two major problems with this approach. One is the frequent poor quality/quantity of DNA extracted from DBSs on filter paper, and the other is the difficulty in designing primer sets or probes to separate allele-specific melting curves. In this study, we addressed these problems and established a rapid, accurate and simple screening system for SMA with PCR-HRMA using DNA extracted from DBSs on filter paper. METHODS: Seventy individuals were assayed in this study, 42 SMA patients and 28 controls, all of whom had been previously been screened for SMA by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) using DNA extracted from freshly collected blood. In this study, the DNA of each individual was extracted from dried blood that had been spotted onto cards and stored at room temperature (20 - 25 degrees C) for between 1 and 8 years. PCR amplification of 30 or 45 cycles was performed using 50 ng of DNA and was immediately followed by HRMA. SMN1 and SMN2 products were co-amplified using a previously designed primer set (R111 and 541C770) containing two single nucleotide differences. RESULTS: The absorbance ratio at 260/280 of DNA extracted from DBSs ranged from 1.49 to 2.1 (mean ± SD; 1.66 ± 0.12), suggesting high-purity DNA. Thirty cycles of PCR amplification were insufficient to amplify the target alleles; PCR with 45 cycles was, however, successful in 69 out of 70 samples. PCR-HRMA using the R111/541C770 primer set enabled separation of the normalized melting curves of the samples with no SMN1 from those with SMN1 and SMN2. CONCLUSIONS: DBSs on filter paper can be a good source of DNA for the diagnosis of diseases and PCR-HRMA using DNA extracted from DBSs is an alternative method to detect the SMN1 deletion. These findings suggest that the SMA screening system using PCR-HRMA with DBSs on filter paper is practicable in a large population study over a long time period.CLIN LAB PUBL, 2015, Clinical laboratory, 61(5-6) (5-6), 575 - 80, English, International magazine[Refereed]Scientific journal
- Post-transplant lymphoproliferative disorder (PTLD) is a well-recognized aggressive disease commonly associated with Epstein-Barr virus (EBV) infection after hematopoietic stem cell transplantation (HSCT). Although rituximab (RTX) is incorporated into the first-line therapy for EBV-PTLD patients, the outcome of the clinically overt disease is still not optimal mainly due to the regrowth of tumor cells. The proliferation of CD20-/CD19+ tumor cells is increasingly reported in RTX-treated EBV-PTLD patients, whereas the emergence of CD20-/CD19- tumor cells is barely recognized. Here, we report a fatal case of an 18-year-old patient who developed EBV-PTLD after allogeneic HSCT for anaplastic large-cell lymphoma. On day 60 after HSCT, the patient developed abdominal pain, watery diarrhea, and low-grade fever. Colon biopsy revealed the proliferation of CD20+/CD19+/EBV-encoded RNA (EBER)+ tumor cells, and an increase of EBV DNA was detected in peripheral blood (PB). He was treated with RTX for EBV-PTLD and was cleared of EBV DNA in PB. However, he manifested high-grade fever, pancytopenia, and elevated soluble interleukin-2 receptor with a prominent hemophagocytosis in bone marrow aspirates and was treated with etoposide for hemophagocytic lymphohistiocytosis (HLH) complication. He then developed EBV DNA positivity in PB and finally died of Bacteroides fragilis sepsis subsequent to bloody stool and ileus on day 163. Autopsy revealed erosion and bleeding in the whole colon with the proliferation of CD20-/CD19-/EBER+ tumor cells. Immunohistochemical analysis uncovered the CD3-/CD56-/CD79a+/CD79b+ B-cell origin of tumor cells. This case clinically demonstrates the removal of both CD20 and CD19 antigens from EBER+ B cells in an RTX-treated EBV-PTLD patient with HLH complication.SPRINGER, Dec. 2014, European journal of pediatrics, 173(12) (12), 1615 - 8, English, International magazine[Refereed]Scientific journal
- BACKGROUND: Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder caused by deletion or intragenic mutation of SMN1. SMA is classified into several subtypes based on clinical severity. It has been reported that the copy number of SMN2, a highly homologous gene to SMN1, is associated with clinical severity among SMA patients with homozygous deletion of SMN1. The purpose of this study was to clarify the genotype-phenotype relationship among the patients without homozygous deletion of SMN1. METHODS: We performed molecular genetic analyses of SMN1 and SMN2 in 112 Japanese patients diagnosed as having SMA based on the clinical findings. For the patients retaining SMN1, the PCR or RT-PCR products of SMN1 were sequenced to identify the mutation. RESULTS: Out of the 112 patients, 106 patients were homozygous for deletion of SMN1, and six patients were compound heterozygous for deletion of one SMN1 allele and intragenic mutation in the retained SMN1 allele. Four intragenic mutations were identified in the six patients: p.Ala2Val, p.Trp92Ser, p.Thr274TyrfsX32 and p.Tyr277Cys. To the best of our knowledge, all mutations except p.Trp92Ser were novel mutations which had never been previously reported. According to our observation, clinical severity of the six patients was determined by the type and location of the mutation rather than SMN2 copy number. CONCLUSION: SMN2 copy number is not always associated with clinical severity of SMA patients, especially SMA patients retaining one SMN1 allele.ELSEVIER SCIENCE BV, Nov. 2014, Brain & development, 36(10) (10), 914 - 20, English, International magazine[Refereed]Scientific journal
- (一社)日本小児血液・がん学会, Oct. 2014, 日本小児血液・がん学会雑誌, 51(4) (4), 233 - 233, English化学療法中に虫垂炎を発症した小児悪性腫瘍症例の検討(Acute appendicitis of cancer children during chemotherapy: A single center experience)[Refereed]
- (一社)日本小児血液・がん学会, Oct. 2014, 日本小児血液・がん学会雑誌, 51(4) (4), 250 - 250, English神経芽腫がん幹細胞の発生・分化における細胞内小胞輸送の制御因子Rab6Bの役割(Role of a membrane traffic regulator Rab6B in the development of cancer stem cells in neuroblastoma)[Refereed]
- (一社)兵庫県医師会, Mar. 2014, 兵庫県医師会医学雑誌, 56(2) (2), 52 - 52, Japanese神経芽腫微小残存病変(MRD)の評価法に関する研究[Invited]
- Spinal muscular atrophy (SMA) is a common neuromuscular disorder with autosomal recessive inheritance, resulting in the degeneration of motor neurons. The incidence of the disease has been estimated at 1 in 6000-10,000 newborns with a carrier frequency of 1 in 40-60. SMA is caused by mutations of the SMN1 gene, located on chromosome 5q13. The gene product, survival motor neuron (SMN) plays critical roles in a variety of cellular activities. SMN2, a homologue of SMN1, is retained in all SMA patients and generates low levels of SMN, but does not compensate for the mutated SMN1. Genetic analysis demonstrates the presence of homozygous deletion of SMN1 in most patients, and allows screening of heterozygous carriers in affected families. Considering high incidence of carrier frequency in SMA, population-wide newborn and carrier screening has been proposed. Although no effective treatment is currently available, some treatment strategies have already been developed based on the molecular pathophysiology of this disease. Current treatment strategies can be classified into three major groups: SMN2-targeting, SMN1-introduction, and non-SMN targeting. Here, we provide a comprehensive and up-to-date review integrating advances in molecular pathophysiology and diagnostic testing with therapeutic developments for this disease including promising candidates from recent clinical trials.WILEY-BLACKWELL, Sep. 2013, Annals of human genetics, 77(5) (5), 435 - 63, English, International magazine[Refereed]Scientific journal
- Minimal residual disease (MRD) is derived from tumor-initiating cells (TICs) and is responsible for tumor relapse. Neuroblastoma is characterized by extreme tumor heterogeneity, and more than half of high-risk patients experience tumor relapse. To overcome tumor heterogeneity and achieve more sensitive detection of MRD, several sets of real-time RT-PCR markers have been reported for MRD monitoring in neuroblastoma patients from different centers. However, these markers vary across centers and are still being validated. In the present study, we validated the ability of 14 commonly used real-time RT-PCR markers to detect MRD based on their expression in neuroblastoma TICs, and we developed a novel MRD detection protocol, which scored the samples as MRD-positive when the expression of one of the 11 real-time RT-PCR markers (CHRNA3, CRMP1, DBH, DCX, DDC, GABRB3, GAP43, ISL1, KIF1A, PHOX2B and TH) exceeded the normal range. By using this protocol, we prospectively monitored MRD in 73 bone marrow (BM), 12 peripheral blood stem cell and 8 peripheral blood samples from 14 neuroblastoma patients treated at a single center. We scored 100, 56, 56 and 57% BM cytology-positive, elevated vanillylmandelic acid (VMA), elevated homovanillic acid (HVA) and elevated neuron-specific enolase (NSE) samples as MRD-positive, respectively. MRD was also positive in 48, 45, 46 and 43% of the BM cytology-negative and normal VMA, normal HVA and normal NSE samples, respectively. These results suggest that the present MRD detection protocol based on the expression of a set of 11 real-time RT-PCR markers in neuroblastoma TICs achieves sensitive MRD monitoring in neuroblastoma patients.Apr. 2013, Oncology reports, 29(4) (4), 1629 - 36, English, International magazine[Refereed]Scientific journal
- (NPO)日本小児血液・がん学会・(NPO)日本小児がん看護学会・(公財)がんの子供を守る会, Nov. 2012, 日本小児血液・がん学会学術集会・日本小児がん看護学会・公益財団法人がんの子どもを守る会公開シンポジウムプログラム総会号, 54回・10回・17回, 291 - 291, Japanese神経芽腫症例におけるmultiple real-time RT-PCR markerを用いた微小残存病変(MRD)解析[Refereed]
- Neuroblastoma is an aggressive pediatric tumor that accounts for 15% of cancer-related deaths in children. More than half of high-risk neuroblastoma patients develop tumor relapse that is lethal in most cases. A small population of tumor-initiating cells (TICs), recently identified from high-risk neuroblastoma patients as spheres, is believed to be responsible for tumor relapse. Rab family small G proteins are essential in controlling membrane traffic and their misregulation results in several cancers. Rab15 was originally isolated as a brain-specific Rab protein regulating the endocytic recycling pathway and was recently identified as a downstream target of the neural transcription factor Atoh1. Previously, we identified two alternatively spliced Rab15 isoforms in neuroblastoma cells and showed a significant correlation between Rab15 expression and neuronal differentiation. As aberrant alternative splicing is intimately associated with an increasing number of cancers, its use as a new diagnostic and/or prognostic biomarker has attracted considerable attention. In the present study, we explored cancer-associated changes of Rab15 alternative splicing in neuroblastoma TICs. We found that Rab15 alternative splicing generated two novel isoforms designated as Rab15(AN2) and Rab15(AN3) in addition to two known isoforms designated as Rab15(CN) and Rab15(AN1). Although both Rab15(AN2) and Rab15(AN3) contained premature termination codons, they were detected in not only neuroblastoma cells but also in normal human tissues. One isoform was predominantly expressed in the brain and testis, while the other isoform was more specifically expressed in the brain. In neuroblastoma, Rab15 isoform balance measured by the Rab15(CN)/Rab15(AN1+AN2+AN3) ratio was significantly decreased in spheres compared to parental cells. These results suggest that Rab15 alternative splicing may serve as a biomarker to discriminate TICs from non-TICs in neuroblastoma.SPANDIDOS PUBL LTD, Jun. 2012, Oncology reports, 27(6) (6), 2045 - 9, English, International magazine[Refereed]Scientific journal
- (一社)日本小児神経学会, May 2012, 脳と発達, 44(Suppl.) (Suppl.), S216 - S216, JapaneseSMN2遺伝子量解析による予測よりも軽症の経過をとった脊髄性筋萎縮症患者に対するプロモーター解析
- OBJECTIVES: A growing number of epidemiological studies have demonstrated that the consumption of green tea inhibits the growth of a variety of cancers. Epigallocatechin gallate (EGCG), the most abundant catechin in green tea, has been shown to have an anti-cancer effect against many cancers. Most cancers are believed to be initiated from and maintained by a small population of tumor-initiating cells (TICs) that are responsible for chemotherapeutic resistance and tumor relapse. In neuroblastoma, an aggressive pediatric tumor that often relapses and has a poor prognosis, TICs were recently identified as spheres grown in a serum-free non-adherent culture used for neural crest stem cell growth. Although EGCG has been reported to induce growth arrest and apoptosis in neuroblastoma cells, its effect on neuroblastoma TICs remains to be defined. METHODS: Gene expression was analyzed by real-time reverse transcription polymerase chain reaction (RT-PCR). The effects of EGCG on cell proliferation, apoptosis, and sphere formation were determined by cell counting, propidium iodide staining, and sphere (>100 μm in diameter) counting, respectively. RESULTS: Neuroblastoma BE(2)-C cells showed increased expression of stem cell markers (nanog homeobox [NANOG] and octamer-binding transcription factor 4 [OCT4]), as well as decreased expression of neuronal differentiation markers (Cu(2+)-transporting ATPase alpha polypeptide [ATP7A] and dickkopf homolog 2 [DKK2]) in spheres grown in serum-free non-adherent culture, compared to parental cells grown in conventional culture. Although EGCG induced growth arrest and apoptosis in the parental cells in a dose-dependent manner, it was not effective against spheres. However, EGCG potently inhibited sphere formation in the BE(2)-C cells. CONCLUSIONS: The present results suggest that EGCG may inhibit the development of TICs in BE(2)-C cells.May 2012, Environmental health and preventive medicine, 17(3) (3), 246 - 51, English, International magazine[Refereed]Scientific journal
- Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder that is caused by loss of the survival motor neuron gene, SMN1. SMA treatment strategies have focused on production of the SMN protein from the almost identical gene, SMN2. Valproic acid (VPA) is a histone deacetylase inhibitor that can increase SMN levels in some SMA cells or SMA patients through activation of SMN2 transcription or splicing correction of SMN2 exon 7. It remains to be clarified what concentration of VPA is required and by what mechanisms the SMN production from SMN2 is elicited. We observed that in two fibroblast cell lines from Japanese SMA patients, more than 1mM of VPA increased SMN2 expression at both the transcript and protein levels. VPA increased not only full-length (FL) transcript level but also exon 7-excluding (Δ7) transcript level in the cell lines and did not change the ratio of FL/Δ7, suggesting that SMN2 transcription was mainly activated. We also found that VPA modulated splicing factor expression: VPA increased the expression of splicing factor 2/alternative splicing factor (SF2/ASF) and decreased the expression of heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1). In conclusion, more than 1mM of VPA activated SMN2 transcription and modulated the expression of splicing factors in our SMA fibroblast cell lines.ELSEVIER SCIENCE BV, Mar. 2012, Brain & development, 34(3) (3), 213 - 22, English, International magazine[Refereed]Scientific journal
- (一社)日本衛生学会, Feb. 2012, 日本衛生学雑誌, 67(2) (2), 329 - 329, Japanese
- (一社)日本衛生学会, Feb. 2012, 日本衛生学雑誌, 67(2) (2), 270 - 270, Japanese茶カテキンの神経芽腫がん幹細胞に対する抗がん作用の検討[Refereed]
- AIM: Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder. It is caused by mutations in the SMN1, and its clinical severity is modified by copy number variations of the SMN2. According to previous studies, deletion of SMN1 exon 7 is the most frequently observed in patients with SMA. Therefore, molecular analyses exploiting this genetic lesion could be beneficial in the diagnosis of SMA. Unfortunately, in many geographical regions, physicians do not have the latest molecular screening technologies at their immediate disposal. Thus, to overcome this issue, we developed an SMA-diagnosing system using dried blood spots (DBS) placed on filter paper to facilitate remote diagnosis. METHODS: In this study, we validate the applicability of DBS on Flinders Technology Associates (FTA) filter paper for detecting SMN1 exon 7 deletions and copy number variations of SMN1 and SMN2. To detect exon 7 deletions in SMN1, polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis was conducted by using DNA extracted from the DBS on FTA filter paper that had been stored at room temperature for a period of up to 4 years. To determine the copy numbers of SMN1 and SMN2, we carried out SYBR green-based real-time PCR by using the same blood specimens. RESULTS: The results obtained from the DBS on FTA filter paper were in complete concordance with those analyses using fresh blood specimens. This indicates that DBS on filter papers is a reliable method for SMA patient detection and carrier screenings. CONCLUSION: The SMA-diagnosing system, combined with the mailing of DBS on filter paper, will be beneficial for patients suffering from neuromuscular disorders in areas with limited or no access to diagnostic facilities with molecular capabilities.MARY ANN LIEBERT, INC, Feb. 2012, Genetic testing and molecular biomarkers, 16(2) (2), 123 - 9, English, International magazine[Refereed]Scientific journal
- (NPO)日本小児がん学会, Nov. 2011, 小児がん, 48(プログラム・総会号) (プログラム・総会号), 341 - 341, Japanese初診時急性白血病様所見を呈した胞巣型横紋筋肉腫の1例[Refereed]
- AIM: Spinal muscular atrophy (SMA) is a well-defined autosomal recessive neuromuscular disorder caused by mutations in the survival motor neuron 1 (SMN1) gene. The most frequently observed mutation is a deletion of exon 7, which has been documented in >95% of SMA patients. A novel technique for detecting mutations known as high-resolution melting analysis (HRMA) has rapidly become the tool of choice for screening pathogenic genetic variants. In the present study, we attempt to validate the applicability of HRMA to the detection of exon 7 deletions and other intragenic mutations in SMN1. RESULTS: Three primer sets were adopted in our HRMA screening for deletion of SMN1 exon 7. In screening attempts utilizing two primer sets, the results of HRMA were not compatible with those obtained by polymerase chain reaction-restriction fragment length polymorphism. Therefore, we applied a modified protocol using revised primer sets, which resulted in an absolute compatibility of results between HRMA and polymerase chain reaction-restriction fragment length polymorphism. With regard to screenings for intragenic mutations in SMN1 exon 3, two primer sets were adopted for use in HRMA. In the initial HRMA screening using the first primer set, we failed to identify any intragenic mutations; however, when using a revised primer set, HRMA successfully detected the presence of a c.275G>C mutation. CONCLUSION: HRMA is a simple but versatile tool to add to the existing arsenal of diagnostic techniques that could aid clinicians/researchers in diagnosing SMA. However, as we demonstrate in the present study, the design and selection of primers is of monumental importance in ensuring the successful application of HRMA to screening for pathogenic variants.MARY ANN LIEBERT INC, Oct. 2011, Genetic testing and molecular biomarkers, 15(10) (10), 677 - 84, English, International magazine[Refereed]Scientific journal
- 金原出版(株), Oct. 2011, 小児科, 52巻, 11号, pp. 1535-1542(11) (11), 1535 - 1542, Japanese[Refereed]
- Neuroblastoma is the most common extracranial solid tumor in children and accounts for 15% of pediatric cancer deaths. Although retinoic acid (RA) is currently used to treat high-risk neuroblastoma patients in the clinic, RA-responsiveness is variable and unpredictable. Since no alterations in the RA-signaling pathway have been found in neuroblastoma cells, molecules correlated with RA-induced differentiation will provide predictive markers of RA-responsiveness for clinical use. The Rab family of small G proteins are key regulators of membrane traffic and play a critical role in cell differentiation and cancer progression. Although an increasing number of cancer-associated alternative splicing events have been identified, alternative splicing of Rab proteins remains to be characterized in neuroblastoma. In the present study, we focused on Rab15 that was originally identified as a brain-specific Rab protein and regulates the endocytic recycling pathway. We identified alternatively spliced Rab15 isoforms designated as Rab15CN and Rab15AN in neuroblastoma cells. Rab15CN was composed of 7 exons encoding 212 amino acids and showed brain-specific expression. Alternative splicing of exon 4 generated Rab15AN that was predicted to encode 208 amino acids and was predominantly expressed in testis. RA induced neuronal differentiation of neuroblastoma BE(2)-C cells and specifically up-regulated Rab15CN expression. Reciprocally, RA-induced differentiation was observed in Rab15CN-expressing BE(2)-C cells in preference to Rab15AN-expressing BE(2)-C cells. Furthermore, Rab15CN expression was also specifically up-regulated during RA-induced differentiation of newly established neuroblastoma cells from high-risk patients. These results suggest that Rab15 expression correlates with RA-induced differentiation of neuroblastoma cells.SPANDIDOS PUBL LTD, Jul. 2011, Oncology reports, 26(1) (1), 145 - 51, English, International magazine[Refereed]Scientific journal
- Licorice ingestion, as well as mutations in the HSD11B2 gene, inhibits 11β-hydroxysteroid dehydrogenase type 2 (11βHSD2) enzyme activity, causing the syndrome of apparent mineral corticoid excess (AME). However, the combined effect of licorice ingestion and an HSD11B2 mutation has never been reported, until now. In this study, we demonstrated that licorice ingestion can produce overt hypertension in an individual without medical history of hypertension who is heterozygous for wild-type and mutant HSD11B2 genes. Our patient was a 51-year-old female with serious hypertension who had been taking herbal medicine containing licorice for more than one year. She was clinically diagnosed as having licorice intoxication, because she did not present with hypertension after ceasing the herbal medicine. Molecular analysis showed that she carried a missense mutation, c.40C>T, in HSD11B2. In conclusion, licorice ingestion is an environmental risk factor for hypertension or AME state in patients with a mutation in HSD11B2. Carrying a mutation in HSD11B2 is, conversely, a genetic risk factor for licorice-induced hypertension or AME state. Herbal medicine containing licorice may, therefore, be contraindicated in patients with an HSD11B2 mutation.HINDAWI PUBLISHING CORPORATION, 2011, Evidence-based complementary and alternative medicine : eCAM, 2011, 646540 - 646540, English, International magazine[Refereed]Scientific journal
- Molecular characterization of the 5'-UTR of retinal dystrophin reveals a cryptic intron that regulates translational activity.PURPOSE: Mutations in the dystrophin (DMD) gene cause Duchenne or Becker muscular dystrophy (DMD/BMD). DMD contains a retina-specific promoter in intron 29. The short R-dystrophin transcript from this promoter has a retina-specific exon 1 (R1) joined to exon 30 of the DMD gene. It has been claimed that this is responsible for the ophthalmological problems observed in DMD/BMD. This research characterizes the structure of the 5'-untranslated region (5'-UTR) of human R-dystrophin. METHODS: The 5'-UTR of the human R-dystrophin transcript was amplified from human retina and 20 other human tissue RNAs by reverse transcription polymerase chain reaction (RT-PCR). Amplified products were identified by sequencing. The translational activities of transcripts bearing differing 5'-UTRs were measured using a dual luciferase assay system. RESULTS: RT-PCR amplification of the R-dystrophin transcript from the retina using a conventional primer set revealed one product comprising exon R1 and exons 30 to 32 (R-dys α). In contrast, three amplified products were obtained when a forward primer at the far 5'-end of exon R1 was employed for RT-PCR. R-dys α, and a shorter form in which 98 bp was deleted from exon R1 (R-dys β), were the two major products. A minor, short form was also identified, in which 143 bp was deleted from exon R1 (R-dys γ). The two primary retinal products (R-dys α and β) encoded an identical open reading frame. The 98 bp deleted in R-dys β was identified as a cryptic intron that was evolutionarily acquired in higher mammals. The shorter R-dys β was expressed in several tissues with a wide range in expression level, while R-dys α was retina specific. The 5'-UTRs of R-dys α and β were examined for translational activity using a dual luciferase assay system. Unexpectedly, the 5'-UTR of R-dys β showed lower translational activity than that of R-dys α. This lower activity was presumed to be due to the removal of internal ribosome entry sites by activation of cryptic intron splicing. CONCLUSIONS: An evolutionarily-acquired cryptic intron was identified in the 5'-UTR of the human R-dystrophin transcript. The two abundant R-dystrophin transcripts in the retina showed different translational activities in vitro owing to their differential splicing of the cryptic intron. This evolutionarily-acquired alternative splicing may act as a molecular switch that regulates translation of the R-dystrophin transcript.MOLECULAR VISION, Dec. 2010, Molecular vision, 16(277-79) (277-79), 2590 - 7, English, International magazine[Refereed]Scientific journal
- (公社)日本小児科学会, Dec. 2010, 日本小児科学会雑誌, 114(12) (12), 1946 - 1946, JapaneseSMN1遺伝子の片側アレルの欠失を認め、もう一方のアレルに点突然変異を認めた脊髄性筋萎縮症の1例
- 金原出版(株), Dec. 2010, 小児科, 51(13) (13), 1797 - 1806, Japanese小児医学最近の進歩 脊髄性筋萎縮症up to date 医学的管理[Refereed]
- 金原出版(株), Nov. 2010, 小児科, 51(12) (12), 1687 - 1695, Japanese小児医学最近の進歩 脊髄性筋萎縮症up to date 遺伝子診断[Refereed]
- 金原出版(株), Oct. 2010, 小児科, 51(11) (11), 1535 - 1542, Japanese小児医学最近の進歩 脊髄性筋萎縮症up to date 遺伝子診断[Refereed]
- Most chromosomal rearrangements including the mixed lineage leukemia (MLL) gene are manifested as leukemia and predict a poor prognosis. Although more than 50 MLL-rearrangement partners are characterized, MLL-related leukemogenesis remains to be understood. Here we report a case of a 3-year old boy bearing a novel MLL-rearrangement with the suppressor of actin mutations 1-like (SACM1L) gene in the absence of leukemia. Bone marrow cells harboring the MLL-SACM1L rearrangement appeared during chemotherapy for acute lymphoblastic leukemia with hyperdiploidy and were continuously detected over 7 years without clonal expansion.PERGAMON-ELSEVIER SCIENCE LTD, Oct. 2010, Leukemia research, 34(10) (10), 1398 - 401, English, International magazine[Refereed]Scientific journal
- Generalized epilepsy with febrile seizures plus (GEFS+) is a childhood genetic epilepsy syndrome. GEFS+ includes a wide spectrum of clinical manifestations, and SCN1A mutations have frequently been reported among the GEFS+-related gene abnormalities. In this study, to clarify the distributions of the clinical subtypes, we analyzed 34 families with GEFS+ in Indonesia using the hospital records of the patients and questionnaires for the family members. The number of patients with febrile seizures plus (FS+), FS+ and afebrile generalized/partial seizures, borderline severe myoclonic epilepsy in infancy (SMEB) and severe myoclonic epilepsy in infancy (SMEI) were 9, 11, 7, and 7, respectively. Most patients had a family history of febrile seizures. Next, we performed molecular analyses to clarify the contributions of SCN1A mutations to the development of the GEFS+ subtypes. Only 3 of 34 probands showed SCN1A mutations. These mutations were two missense mutations, p.V1612I and p.C1756G, in two patients with SMEI and SMEB, and one silent mutation, p.G1762G, in a patient with FS+ and afebrile partial seizures. In conclusion, the majority of GEFS+ patients in Indonesia were not associated with SCN1A mutations. To detect the GEFS+-causing mutations, we must search and analyze other genes in these patients.ELSEVIER SCIENCE BV, Jun. 2010, Epilepsy research, 90(1-2) (1-2), 132 - 9, English, International magazine[Refereed]Scientific journal
- The uridine diphosphoglucuronate-glucuronosyltransferase 1A1 (UGT1A1) gene encodes the enzyme responsible for bilirubin glucuronidation. To evaluate the contribution of UGT1A1 promoter mutations to neonatal jaundice, we determined the genotypes of c.-3279T>G, c.-3156G>A, and A(TA)7TAA in Malay infants with neonatal jaundice (patients) and in infants without neonatal jaundice (controls). In our population study, only c.-3279T>G was associated with neonatal jaundice. The genotype distributions between both groups were significantly different (p = 0.003): the frequency of homozygosity for c.-3279G was much higher in patients than those in controls. Allele frequency of c.-3279G was significantly higher in patients than those in controls (p = 0.006). We then investigated changes in transcriptional activity because of c.-3279T>G. Luciferase reporter assay in HepG2 cells demonstrated that transcriptional activity of the c.-3279G allele was significantly lower than that of the c.-3279T allele in both the absence and presence of bilirubin. Luciferase reporter assay in COS-7 cells elucidated that c.-3279T>G modified the synergistic effects of the nuclear factors associated with transcriptional machinery. In conclusion, the c.-3279T>G mutation in the UGT1A1 promoter is a genetic risk factor for neonatal jaundice.INT PEDIATRIC RESEARCH FOUNDATION, INC, Apr. 2010, Pediatric research, 67(4) (4), 401 - 6, English, International magazine[Refereed]Scientific journal
- 2010, Eastern Journal of Medicine, Vol. 15, No. 4, pp. 155-162, EnglishGenetic disorders associated with neonatal jaundice[Refereed]
- Tight junctions (TJs) and adherens junctions (AJs) comprise epithelial apical junctions that adhere neighboring epithelial cells and determine tissue organization. They are highly dynamic structures that undergo continuous remodeling during physiological morphogenesis and under pathological conditions. The assembly and disassembly of epithelial apical junctions is regulated by the interplay between a variety of cellular processes, such as the remodeling of actin cytoskeletons and the endocytic recycling of apical junctional proteins, and coordinated by many signaling pathways. Accumulating evidences demonstrate that Rab family small G proteins are crucially involved in the regulation of epithelial apical junctions. Rab proteins localized both at endosomes and apical junctions can influence the assembly and disassembly of epithelial apical junctions. In this review, we summarize how Rab proteins influence epithelial apical junctions and describe the role of Rab8/13-a junctional Rab13-binding protein (JRAB)/molecule interacting with CasL-like 2 (MICAL-L2) complexes in the regulation of epithelial apical junctions.FRONTIERS IN BIOSCIENCE INC, Jan. 2009, Frontiers in bioscience, 14, 2115 - 29, English, International magazine[Refereed]Scientific journal
- Rab3 subfamily small G proteins (Rab3A, Rab3B, Rab3C, and Rab3D) control the regulated exocytosis in neuronal/secretory cells. Rab3B is also detected and upregulated in non-neuronal/non-secretory cells, whereas its function remains elusive. In the present study, we identified growth-arrest-specific gene 8 (Gas8), an evolutionally conserved microtubule-binding protein that is upregulated in growth-arrested NIH 3T3 cells and involved in the dynein motor regulation in flagellar/ciliary axoneme, as a novel Rab3B-binding protein using a yeast two-hybrid system. Rab3B as well as Gas8 was upregulated in growth-arrested NIH 3T3 cells and enriched in testis and lung with well-developed flagella/cilia. Gas8 was specifically interacted with the GTP-bound form of Rab3B and co-localized with Rab3B at the Golgi in NIH 3T3 cells. Furthermore, Rab3B was relocated upon expression of the Rab3B-binding domain of Gas8. These results suggest that Gas8 links Rab3B to microtubules in NIH 3T3 cells.ELSEVIER SCIENCE INC, Jun. 2008, Archives of biochemistry and biophysics, 474(1) (1), 136 - 42, English, International magazine[Refereed]Scientific journal
- Tight junctions (TJs) are cell-cell adhesive structures that undergo continuous remodeling. We previously demonstrated that Rab13 and a junctional Rab13-binding protein (JRAB)/molecule interacting with CasL-like 2 (MICAL-L2) localized at TJs and mediated the endocytic recycling of the integral TJ protein occludin and the formation of functional TJs. Here, we investigated how JRAB/MICAL-L2 was targeted to TJs. Using a series of deletion mutants, we found the plasma membrane (PM)-targeting domain within JRAB/MICAL-L2. We then identified actinin-4, which was originally isolated as an actin-binding protein associated with cell motility and cancer invasion/metastasis, as a binding protein for the PM-targeting domain of JRAB/MICAL-L2, using a yeast two-hybrid system. Actinin-4 was colocalized with JRAB/MICAL-L2 at cell-cell junctions and linked JRAB/MICAL-L2 to F-actin. Although actinin-4 bound to JRAB/MICAL-L2 without Rab13, the actinin-4-JRAB/MICAL-L2 interaction was enhanced by Rab13 activation. Depletion of actinin-4 by using small interfering RNA inhibited the recruitment of occludin to TJs during the Ca(2+) switch. During the epithelial polarization after replating, JRAB/MICAL-L2 was recruited from the cytosol to cell-cell junctions. This JRAB/MICAL-L2 recruitment as well as the formation of functional TJs was delayed in actinin-4-depleted cells. These results indicate that actinin-4 is involved in recruiting JRAB/MICAL-L2 to cell-cell junctions and forming functional TJs.AMER SOC MICROBIOLOGY, May 2008, Molecular and cellular biology, 28(10) (10), 3324 - 35, English, International magazine[Refereed]Scientific journal
- Doc2 alpha and Munc13-4 regulate Ca(2+) -dependent secretory lysosome exocytosis in mast cells.The Doc2 family comprises the brain-specific Doc2alpha and the ubiquitous Doc2beta and Doc2gamma. With the exception of Doc2gamma, these proteins exhibit Ca(2+)-dependent phospholipid-binding activity in their Ca(2+)-binding C2A domain and are thought to be important for Ca(2+)-dependent regulated exocytosis. In excitatory neurons, Doc2alpha interacts with Munc13-1, a member of the Munc13 family, through its N-terminal Munc13-1-interacting domain and the Doc2alpha-Munc13-1 system is implicated in Ca(2+)-dependent synaptic vesicle exocytosis. The Munc13 family comprises the brain-specific Munc13-1, Munc13-2, and Munc13-3, and the non-neuronal Munc13-4. We previously showed that Munc13-4 is involved in Ca(2+)-dependent secretory lysosome exocytosis in mast cells, but the involvement of Doc2 in this process is not determined. In the present study, we found that Doc2alpha but not Doc2beta was endogenously expressed in the RBL-2H3 mast cell line. Doc2alpha colocalized with Munc13-4 on secretory lysosomes, and interacted with Munc13-4 through its two regions, the N terminus containing the Munc13-1-interacting domain and the C terminus containing the Ca(2+)-binding C2B domain. In RBL-2H3 cells, Ca(2+)-dependent secretory lysosome exocytosis was inhibited by expression of the Doc2alpha mutant lacking either of the Munc13-4-binding regions and the inhibition was suppressed by coexpression of Munc13-4. Knockdown of endogenous Doc2alpha also reduced Ca(2+)-dependent secretory lysosome exocytosis, which was rescued by re-expression of human Doc2alpha but not by its mutant that could not bind to Munc13-4. Moreover, Ca(2+)-dependent secretory lysosome exocytosis was severely reduced in bone marrow-derived mast cells from Doc2alpha knockout mice. These results suggest that the Doc2alpha-Muunc13-4 system regulates Ca(2+)-dependent secretory lysosome exocytosis in mast cells.AMER ASSOC IMMUNOLOGISTS, Apr. 2008, Journal of immunology, 180(7) (7), 4774 - 84, English, International magazine[Refereed]Scientific journal
- The assembly of tight junctions (TJs) and adherens junctions (AJs) is regulated by the transport of integral TJ and AJ proteins to and/or from the plasma membrane (PM) and it is tightly coordinated in epithelial cells. We previously reported that Rab13 and a junctional Rab13-binding protein (JRAB)/molecule interacting with CasL-like 2 (MICAL-L2) mediated the endocytic recycling of an integral TJ protein occludin and the formation of functional TJs. Here, we investigated the role of Rab13 and JRAB/MICAL-L2 in the transport of other integral TJ and AJ proteins claudin-1 and E-cadherin to the PM by using a Ca(2+)-switch model. Although knockdown of Rab13 specifically suppressed claudin-1 and occludin but not E-cadherin transport, knockdown of JRAB/MICAL-L2 and expression of its Rab13-binding domain (JRAB/MICAL-L2-C) inhibited claudin-1, occludin, and E-cadherin transport. We then identified Rab8 as another JRAB/MICAL-L2-C-binding protein. Knockdown of Rab8 inhibited the Rab13-independent transport of E-cadherin to the PM. Rab8 and Rab13 competed with each other for the binding to JRAB/MICAL-L2 and functionally associated with JRAB/MICAL-L2 at the perinuclear recycling/storage compartments and PM, respectively. These results suggest that the interaction of JRAB/MICAL-L2 with Rab8 and Rab13 coordinates the assembly of AJs and TJs.AMER SOC CELL BIOLOGY, Mar. 2008, Molecular biology of the cell, 19(3) (3), 971 - 83, English, International magazine[Refereed]Scientific journal
- Epithelial cell scattering recapitulates the first steps of carcinoma invasion/metastasis. While the balance between cell-cell adhesive activity and cell motility ultimately determines this process, its molecular mechanisms remain unclear. Adherence junctions and tight junctions (TJs) are primarily responsible for cell-cell adhesive activity and subjected to dynamic remodeling. We previously showed that Rab13 and its effector protein JRAB/MICAL-L2 mediate the endocytic recycling of the integral TJ protein occludin and the assembly of functional TJs. In this study, we examined the role of Rab13 and JRAB/MICAL-L2 in the scattering of Madin-Darby canine kidney (MDCK) cells in response to 12-O-tetradecanoylphorbol-13-acetate (TPA). Knockdown of Rab13 in canine MDCK cells suppressed the TPA-induced scattering, and this phenotype was restored by re-expression of human Rab13. During TPA-induced MDCK cell scattering, Rab13 was transiently activated and returned to its basal level, and both Rab13 and JRAB/MICAL-L2 were colocalized with F-actin at cell-cell contact sites and then accumulated at emerging lamellipodial structures. TPA-induced MDCK cell scattering was also inhibited by knockdown of canine JRAB/MICAL-L2 and rescued by re-expression of mouse JRAB/MICAL-L2. These results indicate that Rab13 and JRAB/MICAL-L2 are involved in epithelial cell scattering.NATURE PUBLISHING GROUP, Mar. 2008, Oncogene, 27(12) (12), 1687 - 1695, English[Refereed]Scientific journal
- A proper balance between cell adhesion and repulsion is essential for cellular morphogenesis during epithelial-mesenchymal transition and mesenchymal-epithelial transition. A number of ligand-receptor pairs including hepatocyte growth factor/scatter factor-Met and semaphorin-plexin are known to control this balance through the complex intracellular signaling pathways. Cell adhesion to other cells and extracellular matrix (ECM) is mediated by cell adhesion molecules (CAMs) and ECM receptors, respectively, which are associated with cytoskeleton through a variety of plaque proteins strengthening and/or weakening adhesion activities. Cell repulsion requires the downregulation of cell adhesion and the extensive changes in cytoskeletal dynamics. The endocytic recycling of CAMs and ECM receptors has recently emerged as an important mechanism to control the balance between cell adhesion and repulsion. Molecule interacting with CasL (MICAL) family proteins are originally identified as a plaque protein associated with ECM receptors integrins and implicated in semaphorin-plexin dependent repulsive axon guidance. We have recently shown that MICAL family protein JRAB/MICAL-L2 functions as an effector protein for Rab family small G protein Rab13 and regulates the endocytic recycling of tight junctional CAM occludin and controls the adhesion and repulsion of epithelial cells.Feb. 2008, The journal of medical investigation : JMI, 55(1-2) (1-2), 9 - 16, English, Domestic magazine[Refereed]Scientific journal
- The Rab family small G proteins are localized to distinct subsets of intracellular membranes and play a key role in membrane traffic through the interaction with their specific effector protein(s). Rab13 is identified as a plaque protein at tight junctions (TJs) and has been shown to regulate the assembly of functional TJs in epithelial cells. We have demonstrated that Rab13 mediates the endocytic recycling of integral TJ protein occludin, and identified a junctional Rab13-binding protein (JRAB)/molecule interacting with CasL-like 2 (MICAL-L2) as a Rab13 effector protein using a yeast two-hybrid system. JRAB/MICAL-L2 has a calponin-homology domain in the N-terminus, a LIM domain in the middle, and a coiled-coil domain at the C-terminus, and specifically binds to the GTP-bound form of Rab13 via its C-terminus. It is localized to TJs in epithelial cells and distributed along stress fibers in fibroblasts. In epithelial cells, JRAB/MICAL-L2 as well as Rab13 mediates the endocytic recycling of occludin, but not transferrin receptor, and the formation of functional TJs. This chapter describes the procedures for the isolation of JRAB/MICAL-L2 and the analysis of its functions.ELSEVIER ACADEMIC PRESS INC, 2008, Methods in enzymology, 438, 141 - 53, English, International magazine[Refereed]Scientific journal
- The dynamic turnover of adherens junctions (AJs) and tight junctions (TJs) is essential for epithelial morphogenesis during normal development and differentiation. Although the endocytic recycling of E-cadherin is characterized and implicated in AJ turnover, the molecular basis for TJ turnover is poorly understood. Occludin and claudins are distinct transmembrane proteins localized to the TJs. Although claudins are an indispensable structural component of TJ strands, depletion of occludin in mice reveals well-developed TJ strands and complex histological abnormalities. To examine the intracellular transport of transmembrane proteins to and from the cell surface, cell-surface biotinylation is a proven powerful method. Using this method, we successfully demonstrated that occludin was endocytosed and recycled back to the cell surface in both fibroblastic baby hamster kidney (BHK) and epithelial MTD-1A cells. The endocytic recycling of occludin as well as the formation of functional TJs was dependent on Rab13 and a junctional Rab13-binding protein (JRAB)/molecule interacting with CasL-like 2 (MICAL-L2). We describe the method to study the intracellular transport of occludin to and from the cell surface in both fibroblastic and epithelial cells.2008, Methods in molecular biology (Clifton, N.J.), 440, 89 - 96, English, International magazine[Refereed]Scientific journal
- The dynamic turnover of tight junctions (TJs) is essential for epithelial-mesenchymal transitions and/or mesenchymal-epithelial transitions during epithelial morphogenesis. We previously demonstrated that Rab13 specifically mediates the endocytic recycling of occludin. Here, we identified MICAL-L2 (molecule interacting with CasL-like 2) as a novel Rab13-binding protein. Immunoprecipitation and immunofluorescence microscopy showed that MICAL-L2 specifically bound to the GTP-bound form of Rab13 via its C terminus, which contained a coiled-coil domain, and localized at TJs in epithelial MTD-1A cells. Recycling assay demonstrated that a MICAL-L2 mutant lacking the Rab13-binding domain (MICAL-L2-N) specifically inhibited the endocytic recycling of occludin but not transferrin receptor. Ca2+ switch assay further revealed that MICAL-L2-N as well as Rab13 Q67L inhibited the recruitment of occludin to the plasma membrane, the development of transepithelial electrical resistance, and the formation of a paracellular diffusion barrier. MICAL-L2 was displaced from TJs upon actin depolymerization and was distributed along radiating actin cables and stress fibers in Ca2+-depleted MTD-1A and fibroblastic NIH3T3 cells, respectively. These results suggest that MICAL-L2 mediates the endocytic recycling of occludin and the formation of functional TJs by linking Rab13 to actin cytoskeleton. We rename MICAL-L2 as JRAB (junctional Rab13-binding protein).AMER SOC CELL BIOLOGY, May 2006, Molecular biology of the cell, 17(5) (5), 2465 - 75, English, International magazine[Refereed]Scientific journal
- 株式会社医学書院, Apr. 2006, 生体の科学, 57(2) (2), 128 - 133, Japanese[Refereed]
- During epithelial morphogenesis, adherens junctions (AJs) and tight junctions (TJs) undergo dynamic reorganization, whereas epithelial polarity is transiently lost and reestablished. Although ARF6-mediated endocytic recycling of E-cadherin has been characterized and implicated in the rapid remodeling of AJs, the molecular basis for the dynamic rearrangement of TJs remains elusive. Occludin and claudins are integral membrane proteins comprising TJ strands and are thought to be responsible for establishing and maintaining epithelial polarity. Here we investigated the intracellular transport of occludin and claudins to and from the cell surface. Using cell surface biotinylation and immunofluorescence, we found that a pool of occludin was continuously endocytosed and recycled back to the cell surface in both fibroblastic baby hamster kidney cells and epithelial MTD-1A cells. Biochemical endocytosis and recycling assays revealed that a Rab13 dominant active mutant (Rab13 Q67L) inhibited the postendocytic recycling of occludin, but not that of transferrin receptor and polymeric immunoglobulin receptor in MTD-1A cells. Double immunolabelings showed that a fraction of endocytosed occludin was colocalized with Rab13 in MTD-1A cells. These results suggest that Rab13 specifically mediates the continuous endocytic recycling of occludin to the cell surface in both fibroblastic and epithelial cells.AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Jan. 2005, The Journal of biological chemistry, 280(3) (3), 2220 - 8, English, International magazine[Refereed]Scientific journal
- Lead, (株)先端医学社, Jun. 2004, G.I.Research, 12(3) (3), 242 - 248, Japanese細胞内小胞輸送のしくみ[Refereed]
- The Rab3 family small G proteins (Rab3A-D) are involved in the regulated secretory pathway of brain and secretory tissues. Among Rab3-interacting proteins, Rabphilin-3, Rim, and Noc2, all of which contain a conserved Rab3-binding domain (RBD3), are generally recognized Rab3 effector proteins in neurons and secretory cells. Although Rab3B was also detected in epithelial cells, its function remained unknown. We isolated cDNA sequences from human epithelial Caco2-cell mRNA by degenerate RT-PCR based on the conserved amino acid sequence of RBD3. Multiple cDNA clones were identified as encoding Noc2. Northern blot analysis revealed that Noc2 mRNA was expressed not only in secretory tissues but also in epithelial tissues and cell lines. A pull-down assay demonstrated that Noc2 bound to Rab3B in a GTP-dependent manner. When Noc2 was co-expressed with the GTP-bound form of Rab3B, it was recruited from the cytosol to perinuclear membranes. Furthermore, overexpression of Noc2 inhibited the cell-surface transport of basolateral vesicular stomatitis virus glycoprotein. These results suggest that Noc2 functions as a potential Rab3B effector protein in epithelial cells.ACADEMIC PRESS INC ELSEVIER SCIENCE, Mar. 2004, Biochemical and biophysical research communications, 316(1) (1), 218 - 25, English, International magazine[Refereed]Scientific journal
- 一般社団法人日本外科学会, Mar. 2004, 日本外科学会雑誌, 105, 472 - 472, JapanesePS-086-1 Rabファミリー低分子量G蛋白質Rab13によるタイトジャンクションの接着分子Occludinの小胞輸送制御機構
- (株)羊土社, Sep. 2003, 実験医学, 21(14) (14), 2032 - 2038, Japanese【細胞内輸送研究の最前線】 細胞内輸送の異常と疾患 シナプス可塑性を支える小胞輸送の制御機構[Refereed]
- Regulated transport of proteins to distinct plasma membrane domains is essential for the establishment and maintenance of cell polarity in all eukaryotic cells. The Rab family small G proteins play a crucial role in determining the specificity of vesicular transport pathways. Rab3B and Rab13 localize to tight junction in polarized epithelial cells and cytoplasmic vesicular structures in non-polarized fibroblasts, but their functions are poorly understood. Here we examined their roles in regulating the cell-surface transport of apical p75 neurotrophin receptor (p75NTR), basolateral low-density lipoprotein receptor (LDLR), and tight junctional Claudin-1 using transport assay in non-polarized fibroblasts. Overexpression of Rab3B mutants inhibited the cell-surface transport of LDLR, but not p75NTR and Claudin-1. In contrast, overexpression of Rab13 mutants impaired the transport of Claudin-1, but not LDLR and p75NTR. These results suggest that Rab3B and Rab13 direct the cell-surface transport of LDLR and Claudin-1, respectively, and may contribute to epithelial polarization.ACADEMIC PRESS INC ELSEVIER SCIENCE, Aug. 2003, Biochemical and biophysical research communications, 308(2) (2), 270 - 5, English, International magazine[Refereed]Scientific journal
- 徳島医学会, Apr. 2003, 四国医学雑誌, 59(1〜2) (1〜2), 2 - 6, Japanese【トランスレーショナルリサーチ ここまで近づいた基礎研究と臨床の現場】 徳島発の臨床応用を目指した基礎研究 トランスレーショナルリサーチを視野にいれた「小胞輸送」研究[Refereed]Scientific journal
- Presenilin-binding protein forms aggresomes in monkey kidney COS-7 cells.A novel presenilin-binding protein (PBP) is specifically expressed in the brain, and its level in the soluble fraction of Alzheimer's disease (AD) brains is much less than that in the age-matched controls. Recently, several proteins, including presenilin (PS), have been found to form structures of aggregated proteins, called aggresomes, when the production of the proteins exceeds their rate of degradation by proteasomes. Based on these observations it has been proposed that the aggresome may represent one of the mechanisms forthe formation of cytoplasmic deposits which are linked to the pathogenesis of neurodegenerative disorders including AD. It is shown here that the overexpression of PBP or the suppression of proteasome activity in monkey kidney COS-7 cells leads to the accumulation of detergent-insoluble and multiubiquitinated PBP aggregates. PBP also forms aggregates in primary cultures of neurons in the presence of a proteasome inhibitors. PBP aggregates have the characteristics of aggresomes, including the localization to microtubule organization centers and the disruption of intermediate filaments. These observations suggest that the malfunctioning of the proteasome can cause the formation of PBP aggresomes, which may lead to AD.Aug. 2002, Journal of neurochemistry, 82(4) (4), 819 - 27, English, International magazine[Refereed]Scientific journal
- The delta subunit of AP-3 is required for efficient transport of VSV-G from the trans-Golgi network to the cell surface.Vesicular stomatitis virus glycoprotein (VSV-G) is a transmembrane protein that functions as the surface coat of enveloped viral particles. We report the surprising result that VSV-G uses the tyrosine-based di-acidic motif (-YTDIE-) found in its cytoplasmic tail to recruit adaptor protein complex 3 for export from the trans-Golgi network. The same sorting code is used to recruit coat complex II to direct efficient transport from the endoplasmic reticulum to the Golgi apparatus. These results demonstrate that a single sorting sequence can interact with sequential coat machineries to direct transport through the secretory pathway. We propose that use of this compact sorting domain reflects a need for both efficient endoplasmic reticulum export and concentration of VSV-G into specialized post-trans-Golgi network secretory-lysosome type transport containers to facilitate formation of viral coats at the cell surface.May 2002, Proceedings of the National Academy of Sciences of the United States of America, 99(10) (10), 6755 - 60, English, International magazine[Refereed]Scientific journal
- American Society for Biochemistry & Molecular Biology (ASBMB), May 1999, Journal of Biological Chemistry, 274(22) (22), 15937 - 15946[Refereed]Scientific journal
- American Society for Biochemistry & Molecular Biology (ASBMB), Oct. 1998, Journal of Biological Chemistry, 273(41) (41), 26931 - 26938[Refereed]Scientific journal
- Elsevier BV, Jan. 1998, Trends in Cell Biology, 8(1) (1), 21 - 25[Refereed]Scientific journal
- Lead, American Association for the Advancement of Science (AAAS), Jul. 1997, Science, 277(5325) (5325), 556 - 558[Refereed]Scientific journal
- The p21 protein inhibits the activity of cyclin-Cdk complexes and suppresses cell cycle progression. Wild type p53 can induce p21, but mutated p53 cannot. Previous studies have demonstrated that mutation of p53 is absent in neuroblastoma (NB). These reports prompted us to examine whether p53 induced p21 in NB. We examined the expression of p21 and p53 mRNA in eight NB, two Ewing's sarcoma (ES) and two primitive neuroectodermal tumor (PNET) cell lines by Northern blot analysis, and sequenced p53 cDNA of these cells. Although p53 mRNA was detected in all analyzed cell lines by Northern blot analysis, p21 mRNA was detected in six NB but not in two NB, two ES and two PNET cell lines. We detected the point mutation of p53 at codon 273 (CGT to TGT) in one NB and two ES cell lines. The non-transforming substitution at codon 72 (CCC to CGC) was detected in all analyzed cell lines. One PNET cell line had a large deletion of p53 cDNA. These results showed that p21 mRNA was usually expressed in NB but not in ES and PNET. This may suggest that the down stream of the p53 signal transduction pathway in NB is different from that of the closely related tumors of ES and PNET. 1997 Japan Pediatric Society.Wiley, 1997, Pediatrics International, 39(5) (5), 590 - 594, English[Refereed]Scientific journal
- Cloning of a Brain-type Isoform of Human Rab GDI and Its Expression in Human Neuroblastoma Cell Lines and Tumor SpecimensRab proteins, a family of Ras-related small GTP-binding proteins, play a key role in regulating intracellular vesicle trafficking. Rab GDP dissociation inhibitor (GDI3) forms a soluble complex with Rab proteins and thereby prevents the exchange of GDP for GTP. Recently, two isoforms of Rab GDI cDNA were isolated from rats and mice. In this study, we have isolated a brain-type isoform of human Rab GDI cDNA and examined its expression in neuroblastoma. We tentatively designate it as human Rab GDI a (hu GDI a) and another human Rab GDI, as human Rab GDI 0 (hu GDI β). Hu GDI a cDNA encodes a protein of 447 amino acids with a deduced molecular weight of 50,200. Northern blot analysis revealed that hu GDI a gene is expressed abundantly in the brain but much less in other tissues, while hu GDI 0 gene is ubiquitously expressed. All human neuroblastoma cell lines and tumor specimens examined express hu GDI a gene to various extents, while a human T cell leukemia cell line, MOLT3, does not The levels of both hu GDI a and 0 mRNA were constant in a human neuroblastoma cell line, NB1, during its neuronal differentiation, while Rab3A and neurofilament-L gene expression and the number of neurosecretory granules were elevated at this condition. These results suggest that hu GDI a gene expression is not related to the differentiation state of neuronal cells. © 1995, American Association for Cancer Research. All rights reserved.Lead, Nov. 1995, Cancer Research, 55(22) (22), 5445 - 5450, English[Refereed]Scientific journal
- Phosphodiesterase I/nucleotide pyrophosphatase is a widely expressed membrane-bound enzyme that cleaves diester bonds of a variety of substrates. We have cloned brain-type cDNA for this enzyme from rat brain and designated it PD-Iα (M. Narita, J. Goji, H. Nakamura, and K. Sano, 1994, J. Biol. Chem. 269: 28235-28242). In this study we have isolated cDNA and genomic DNA encoding human PD-Iα. Human PD-Iα cDNA, designated PDNP2 in HGMW nomenclature, has a 2589-nucleotide open reading frame encoding a polypeptide of 863 amino acids with a calculated M(r) of 99,034. Northern blot analysis revealed that human PD-Iα transcript was present in brain, lung, placenta, and kidney. The database analysis showed that human PD-Iα was identical with human autotaxin (ATX), a novel tumor motility-stimulating factor, except that human PD-Iα lacks 156 nucleotides and 52 amino acids of human ATX. Human PD- Iα and human ATX are likely to be alternative splicing products from the same gene. The 5' region of the human PDNP2 gene contains four putative binding sites of transcription factor Sp1 without typical TATA or CAAT boxes, and there is a potential octamer binding motif in intron 2. From the results of fluorescence in situ hybridization, the human PDNP2 gene is located at chromosome 8q24.1. © 1995 Academic Press. All rights reserved.Elsevier BV, 1995, Genomics, 30(2) (2), 380 - 384, English[Refereed]Scientific journal
- Molecular cloning and characterization of two rab GDI species from rat brain: Brain-specific and ubiquitous typesRab GDP dissociation inhibitor (GDI) is a regulatory protein for Rab proteins that controls not only the GDP/GTP exchange reaction of Rab proteins but also their translocation between the cytosol and cell membranes. Recently, rab GDI cDNAs have been isolated from human, bovine, rat, and Drosophila, and these Rab GDI proteins indicated an important role in the regulation of intracellular vesicle traffic. In this study, we have isolated two different rab GDI cDNAs, designated rat rab GDI α and β, from a rat brain cDNA library using bovine rab GDI cDNA as a probe. Rat rab GDI α and β show 99 and 86% amino acid identity with bovine rab GDI, respectively, indicating that rat rab GDI α is a rat counterpart of bovine rab GDI. Both rat Rab GDI α and β proteins expressed in Escherichia coli showed a similar degree of activity of regulating the GDP/GTP exchange reaction to that of bovine Rab GDI using Rab3A and Rab11 as substrates. Northern blot analysis revealed that rab GDI α mRNA was expressed abundantly in brain but much less in other tissues, whereas rat rab GDI β mRNA was ubiquitously expressed. Reverse transcription-polymerase chain reaction analysis revealed that astrocytes expressed rat rab GDI β gene but not rat rab GDI α gene. These results indicate that Rab GDI α is involved in sorting of highly specialized vesicles in brain such as neurosecretory vesicles, whereas Rab GDI β plays a general role in vesicular trafficking in diverse types of cells.Lead, May 1994, Journal of Biological Chemistry, 269(19) (19), 14191 - 14198, English[Refereed]Scientific journal
- (公社)日本小児科学会, May 1992, 日小児会誌, 96(5) (5), 1270 - 1273, JapaneseDonarth-Landsteiner 抗体が IgM に属し抗I特異性を示した発作性寒冷血色素尿症の1例[Refereed]
- 1児にWilson-Mikity症候群,他の2児に異なる先天奇形を伴った超未熟児の五胎例排卵誘発剤投与により妊娠した母親から出生し,第1子がcongenital cystic adenomatoid malfomation of the lung (CCAM),第2子がWilson-Mikity症候群,第4子がMalrotation Meconium ileus, Meconium peritonitisを呈した超未熟児の五胎例を経験した.5児ともに死亡するという不幸な転帰となったが,1) 5児中2児にCCAMとMalrotationという異なる2種類の先天奇形をみたこと,2) 5児中1児のみに高IgM血症を伴うWilson Mikity症候群をみたこと等の点で注目された(一社)日本周産期・新生児医学会, Mar. 1992, 日本新生児学会雑誌, 28(1) (1), 120 - 125, Japanese[Refereed]
- We measured the platelet count (Plt, mean platelet volume (MPV), platelet distribution width (PDW), platletcrit (Pct), RBC distribution width (RDW) and serum antiepileptic drug levels in 152 outpatients attending our neurology clinic (45 patients receiving VPA monotherapy, 34 receiving PB monotherapy, 13 CBZ monotherapy, 49 nonmedicated), and obtained MPV nomograms.JAPAN EPILEPSY SOCIETY, Mar. 1992, Journal of the Japan Epilepsy Society, 10(1) (1), 28 - 33, Japanese
In patients given VPA monotherapy, we recognized the following features:
1) Statistically significant (p<0.01) decrease of Plt.
2) Statistically significant (p<0.01) increase of MPV.
3) Deviation of MPV nomogram.
There was no correlation between Plt, MPV or serum level and the duration of VPA medication. PDW and Pct showed no change.
In patients given PB and CBZ monotherapy, there was no change in Plt, MPV, PDW, Pct or the MPV nomogram. 当科神経外来通院中の患児142例 (VPA単剤服用中の患児: 45例, PB単剤服用中の患児: 34例, CBZ単剤服用中の患児: 14例, 抗けいれん剤を服用していない患児: 49例) を対象に血小板数 (Plt), 平均血小板容積 (MPV), 血小板分布幅 (PDW), 血小板クリット (Pct), 赤血球分布幅 (RDW), 各抗けいれん剤血中濃度を測定し, MPV nomogram を作成した。
VPA単剤服用患児において,
1) Pltの有意 (p<0.01) な減少
2) MPVの有意 (p<0.01) な増加
3) MPV nomogramの偏位
を認めたが, 血中濃度および投与期間との間に相関関係はなく, PDW, Pctの有意な変化はみられなかった.
PBおよびCBZ単剤服用患児では, Plt, MPV, PDW, Pctの有意な変化はみられなかった。[Refereed] - Fibroblast growth factor (FGF) plus insulin induced DNA synthesis in and proliferation of NIH/3T3 cells. The protein kinase C-activating phorbol ester, 12-o-tetradecanoylphorbol 13-acetate (TPA), inhibited both the DNA synthesis and cell proliferation induced by FGF plus insulin. The concentration of TPA required for50% inhibition of the DNA synthesis was about 5 nM. Phorbol-12,13-dibutyrate, another protein kinase C-activating phorbol ester, also inhibited the DNA synthesis but 4α-phorbol-12,13-didecanoate, known to be inactive for this enzyme, was ineffective. DNA synthesis started at about 12 h after the addition of FGF plus insulin. The inhibitory action of TPA on the DNA synthesis was observed when it was added within 12 h after the addition of FGF plus insulin. These results suggest that phorbol esters exhibit an antiproliferative action through protein kinase C activation in NIH/3T3 cells, and that this action of phorbol esters is due to inhibition of the progression from the late G1 to the S phase of the cell cycle. © 1988 BY THE JOURNAL OF BIOCHEMISTRY.Oxford University Press, 1988, Journal of Biochemistry, 104(1) (1), 53 - 56, English[Refereed]Scientific journal
- (公社)日本小児保健協会, May 2023, 小児保健研究, 82(講演集) (講演集), 108 - 108, Japanese
- 本研究の目的は,新型コロナウイルス感染症(COVID-19)流行下における日本国内の一市中病院小児科外来のワクチン接種児数の変化を明らかにし,その要因と対策を考察することである。本研究では対象機関における小児のワクチン11種類に注目し,2016年～2019年のワクチン接種児数の平均と2020年のワクチン接種児数とを比較した。COVID-19流行後の1年間を通してみると,ロタウイルス,B型肝炎,Hib,肺炎球菌,DPT・IPV,MR,水痘,日本脳炎,DTの9種類のワクチン接種児数は減少し,インフルエンザ,ムンプスの2種類のワクチン接種児数は増加した。COVID-19流行初期の報告では,乳児に比して年長児でより顕著にワクチン接種児数が減少していたが,本研究では乳児でより顕著な減少が認められ,児の年齢が上がるにつれて減少幅は縮小した。これには,本研究の対象機関がCOVID-19患者を診療しており,乳児の保護者が病院受診によるCOVID-19罹患・重症化をより強く懸念していることが影響したと考えた。本研究で接種児数が増加したインフルエンザワクチンとムンプスワクチンには,対象機関のある自治体が独自の助成制度を新設していた。ワクチン接種児数の変化につながる要因の解明とそれに基づいた対策の実施には,医療・保健・福祉・教育・行政機関の連携・協働が益々重要になる。(著者抄録)(公社)日本小児保健協会, May 2023, 小児保健研究, 82(3) (3), 304 - 312, Japanese
- Down症児の感覚と発達特性について検討した.当院にて「ダウン症児の赤ちゃん体操」を行い,歩行の完成を認めた33人の児の保護者に対し,「日本版感覚プロファイル(SP)」と発達特性をみる「日本語版乳幼児自閉症チェックリスト(M-CHAT)」の記入を依頼し,17人から結果を得た.SPでは低登録が「高い」以上の割合が有意に多く(p<0.001),Down症児の感覚閾値の高さと受動的行動が示唆された.感覚処理項目では,口腔感覚で41%の児が「高い」以上を示した(p=0.041).感覚探求が「高い」以上の児では歩行開始年齢が有意に高く,歩行開始年齢が遅れる傾向が明らかになった.M-CHATのスクリーニング陽性者は8人(47.1%)で,陽性者と陰性者の間で,SPの結果に有意な差はみられなかった.Down症児の療育を進めていくうえで個々の感覚特性に見合った対応を考えていく必要がある.(著者抄録)(一社)日本小児精神神経学会, Apr. 2023, 小児の精神と神経, 63(1) (1), 73 - 80, Japanese
- (公社)日本小児科学会, Apr. 2023, 日本小児科学会雑誌, 127(4) (4), 625 - 625, Japanese汎血球減少と斜指を契機に診断したMECOM関連症候群の女児
- (公社)日本小児科学会, Apr. 2023, 日本小児科学会雑誌, 127(4) (4), 632 - 632, Japanese血性鼻汁をきっかけに診断したLCHの男児
- グルコース6リン酸脱水素酵素(G6PD)欠損症は赤血球酵素異常の中で最も頻度が高く,世界で4億人の患者がいるとされるが本邦での有病率は0.1%未満と少ない。感染症や特定の薬剤投与での酸化ストレスにより溶血発作が生じる。症例は当院でフォロー中のG6PD欠損症の18歳男性で,受診2日前から発熱し,受診当日の朝,排尿中に意識消失し自宅で倒れているところを発見され当院に救急搬送された。SARS-CoV-2陽性,Hgb6.6g/dL,LDH 1,894U/L,ハプトグロビン7mg/dLからCOVID-19に伴う溶血発作と診断し赤血球輸血を行った。輸血後全身状態は改善するも,その後も貧血は進行し解熱するまでの間に計6単位の赤血球輸血を要した。我々が知る限り,本症例が本邦でCOVID-19を契機に溶血発作を生じた初の報告である。G6PD欠損症患者は溶血発作,血栓傾向など重症化リスクが高いためCOVID-19発症時には慎重な対応が重要である。(著者抄録)兵庫県小児科医会, 2023, 兵庫県小児科医会報, (79) (79), 13 - 17, Japanese
- (一社)日本輸血・細胞治療学会, Apr. 2022, 日本輸血細胞治療学会誌, 68(2) (2), 261 - 261, Japanese
- (公社)日本小児科学会, Mar. 2022, 日本小児科学会雑誌, 126(3) (3), 552 - 552, Japanese神戸市東部療育センター診療所の開所後3年間のまとめ
- 日本家族看護学会, Sep. 2021, 日本家族看護学会学術集会プログラム・抄録集, 28回, 55 - 55, Japanese特別支援学校に通うことで児童生徒の家族に生じる体験に関する研究
- (一社)日本血液学会, Sep. 2021, 日本血液学会学術集会, 83回, PS - 2, English当科における小児造血細胞移植患者の精子保存(The sperm preservation in pediatric patients who underwent hematopoietic stem cell transplantation)
- (一社)日本家族看護学会, Sep. 2021, 日本家族看護学会学術集会プログラム・抄録集, 28回, 55 - 55, Japanese特別支援学校に通うことで児童生徒の家族に生じる体験に関する研究
- (一社)日本血液学会, Sep. 2021, 日本血液学会学術集会, 83回, PS - 2, EnglishThe sperm preservation in pediatric patients who underwent hematopoietic stem cell transplantation(和訳中)
- (一社)日本輸血・細胞治療学会, May 2021, 日本輸血細胞治療学会誌, 67(2) (2), 307 - 307, Japanese
- (公社)日本小児科学会, Apr. 2021, 日本小児科学会雑誌, 125(4) (4), 681 - 681, Japanese多発乳児血管腫に対するプロプラノロール投与
- (公社)日本小児科学会, Feb. 2021, 日本小児科学会雑誌, 125(2) (2), 250 - 250, English高リスク神経芽腫における微小残存病変(MRD)と腫瘍マーカーの相関に関する臨床的検討(Clinical analysis of the correlation between minimal residual disease and tumor markers in high-risk neuroblastoma)
- (公社)日本小児科学会, Jun. 2020, 日本小児科学会雑誌, 124(6) (6), 1028 - 1028, Japanese予防接種の際に顔面蒼白を指摘されHb2.3g/dLであった1歳6ヵ月女児
- (公社)日本小児科学会, Jun. 2020, 日本小児科学会雑誌, 124(6) (6), 1035 - 1035, Japanese最近当科で経験した上咽頭癌の2例
- 産業開発機構(株), May 2020, 映像情報Medical, 52(5) (5), 40 - 45, Japanese
- WILEY, Dec. 2019, PEDIATRIC BLOOD & CANCER, 66, S50 - S51, EnglishThree Cases of Nasopharyngeal Carcinoma in a Single InstituteSummary international conference
- WILEY, Dec. 2019, PEDIATRIC BLOOD & CANCER, 66, S81 - S82, EnglishLevel of Seven Neuroblastoma-Associated mRNAs Analyzed by ddPCR is Correlated Between Bone Marrow and Peripheral Blood in High-risk NeuroblastomaSummary international conference
- (一社)日本小児精神神経学会, Oct. 2019, 小児の精神と神経, 59(3) (3), 269 - 270, Japanese修正18〜24ヵ月のNICU退院児における感覚特性の検討
- WILEY, Nov. 2018, PEDIATRIC BLOOD & CANCER, 65, S38 - S39, EnglishQuantitation of Minimal Residual Disease by Droplet Digital PCR in High-Risk Neuroblastoma PatientsSummary international conference
- AMER ASSOC CANCER RESEARCH, Oct. 2018, CANCER RESEARCH, 78(19) (19), EnglishSummary international conference
- 2018, 日本小児科学会雑誌, 122(4) (4)乳幼児感覚プロファイルを用いた発達障害児の感覚特性に関する検討
- 2017, 日本小児精神神経学会プログラム・抄録集, 117th, 49, Japanese乳幼児期における発達障害特性と感覚特性との関連についての検討
- (一社)日本周産期・新生児医学会, Jun. 2016, 日本周産期・新生児医学会雑誌, 52(2) (2), 643 - 643, Japanese臍帯由来間葉系幹細胞を用いた脊髄性筋萎縮症の病態解析の試み
- 2016, 日本小児血液・がん学会雑誌(Web), 53(4) (4)PET-MRI所見に基づく肺・下鼻甲介・腎生検が原疾患の診断に有効であった二次性血球貪食性リンパ組織球症の1例
- 2016, 日本小児血液・がん学会雑誌(Web), 53(4) (4)G-CSF投与と自家末梢血幹細胞移植を併用しbi-weekly VDC-IE療法を完遂し得た切除不能ユーイング肉腫例
- (一社)日本臓器保存生物医学会, Oct. 2015, Organ Biology, 22(3) (3), 43 - 43, Japanese
- (一社)日本新生児成育医学会, Sep. 2015, 日本新生児成育医学会雑誌, 27(3) (3), 579 - 579, JapaneseUBアナライザーとビリルビン誘導蛍光タンパク質(UnaG)を用いた新たなアンバウンドビリルビン測定法の開発
- ELSEVIER SCI LTD, Sep. 2015, EUROPEAN JOURNAL OF CANCER, 51, S20 - S20, EnglishContribution of cancer-associated fibroblasts and M2-polarized macrophages to neuroblastoma developmentSummary international conference
- AMER SOC HEMATOLOGY, Dec. 2014, BLOOD, 124(21) (21), EnglishAbsolute Lymphocyte Counts at the End of Induction Is a Prognostic Indicator in Childhood Acute Lymphoblastic LeukemiaSummary international conference
- AMER ASSOC CANCER RESEARCH, Oct. 2014, CANCER RESEARCH, 74(19) (19), EnglishSummary international conference
- AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Sep. 2005, JOURNAL OF BIOLOGICAL CHEMISTRY, 280(36) (36), 32048 - 32048, EnglishRab13 mediates the continuous endocytic recycling of occludin to the cell surface. (vol 280, pg 2220, 2005)Others
- 日本癌学会, 25 Aug. 2003, 日本癌学会総会記事, 62nd, 305 - 305, JapaneseRabファミリー低分子量G蛋白質によるタイトジャンクション膜蛋白質の小胞輸送の制御機構
- 徳島医学会, Apr. 2003, 四国医学雑誌, 59(1〜2) (1〜2), 110 - 111, JapaneseRab3BとRab13低分子量G蛋白質によるゴルジ体 細胞膜間小胞輸送の制御機構
- A 10-month-old male infant (case 1) and another male infant aged 1 year and 11 months (case 2) were admitted to our department because of fever, watery diarrhea and convulsion. On admission, they were unconscious and showed rigidity of the limbs. Laboratory examination revealed a marked increase in GOT and GPT, a decrease in platelet and antithrombin III and an increase in FDP. Metabolic acidosis was found by blood gas analysis. Brain CT showed an extensive area of low density in case 1, and low density centering on the cerebral basal ganglia and brainstem in case 2. Rotavirus was detected in case 2 by fecal examination. The clinical pictures in these cases closely resembled those of hemorrhagic shock and encephalopathy (HSE) reported by Levin et al. in 1983. The etiology of this disease is currently unknown, and its prognosis in poor. The relationship between this disease and rotavirus should be examined in future studies. 症例1は10カ月の男児, 症例2は1歳11カ月の男児.ともに発熱, 水様性下痢, 痙攣を主訴に当科に入院した.入院時, 意識なく四肢硬直していた.検査所見では, GOT, GPTが著明に上昇し, また血小板の減少, fibrinogen degradation products (FDP) の上昇, アンチトロンビンIIIの低下を認めた.血液ガス分析で代謝性アシドーシスを呈した.頭部CTにて症例1では広範囲にわたって低吸収領域を, 症例2では大脳基底核, 脳幹部を中心に低吸収領域を認めた.症例2の糞便検査にてロタウイルスを証明した.この臨床像は, 1983年にLevinらが提唱したhemorrhagic shock and encephalopathy (HSE) と酷似した.現在のところ原因不明であり予後不良の疾患であるがロタウイルスとの関係について今後検討を重ねていく必要があると考えられた.THE JAPANESE SOCIETY OF CHILD NEUROLOGY, 1992, NO TO HATATSU, 24(1) (1), 71 - 77, Japanese
- 第30回日本疫学会学術総会, Feb. 2020, Japanese先天性サイトメガロウィルス感染は自閉症スペクトラム障害のリスク因子か？Poster presentation
- 第61回日本小児血液・がん学会学術集会, Nov. 2019, JapaneseLevel of seven neuroblastoma-associated mRNAs analyzed by droplet digital PCR is correlated between bone marrow and peripheral blood in high-risk neuroblastomaOral presentation
- 第61回日本小児血液・がん学会学術集会, Nov. 2019, JapaneseSpontaneously developed undifferentiated sarcoma and rhabdomyosarcoma in mdx micePoster presentation
- 第61回日本小児血液・がん学会学術集会, Nov. 2019, JapaneseThree cases of nasopharyngeal carcinoma in a single instituteOral presentation
- 第78回日本癌学会学術総会, Sep. 2019, EnglishExpression of seven neuroblastoma-associated mRNAs is correlated between bone marrow and peripheral blood in high-risk neuroblastoma patientsOral presentation
- 第121回日本小児精神神経学会, Jun. 2019, Japanese修正18～24ヶ月のNICU退院児における感覚特性の検討Oral presentation
- 122回日本小児科学会学術集会, Apr. 2019, Japanese小児術前検査における凝固異常の頻度Poster presentation
- 第122回日本小児科学会学術集会, Apr. 2019, Japanese化学療法中に発症した水腎症に対しDouble-Jカテーテル留置を要したALL男児Poster presentation
- 第124回日本解剖学会, Mar. 2019, Japanese臍帯組織由来Muse細胞の栄養膜細胞への分化の可能性Oral presentation
- 第18回日本再生医療学会総会, Mar. 2019, Japanese臍帯組織由来Muse細胞の栄養膜細胞への分化Oral presentation
- 第41回日本造血細胞移植学会総会, Mar. 2019, Japanese移植後の混合キメラに対してデキサメサゾン投与が奏効した家族性血球貪食リンパ組織球症3型の1例Poster presentation
- 13th St. Jude-VIVA Forum in Pediatric Oncology, Feb. 2019, EnglishThe Retrospective Analysis of High-Dose Chemotherapy and Autologous Stem Cell Transplantation for High-Risk Padiatric OsteosarcomaPoster presentation
- 第276回日本小児科学会兵庫県地方会, Feb. 2019, Japanese予防接種の際に顔面蒼白を指摘されHb2.3g/dLであった1歳6か月女児Oral presentation
- ASCB/EMBO 2018 meeting, Dec. 2018, English, San Diego, International conferenceDMD transcription profiling in spontaneously developed tumor from three mdx mice revealed extensive intron retentions and Dp71 isoform expressionPoster presentation
- 第60回日本小児血液・がん学会学術集会, Nov. 2018, English, 京都市, Domestic conferenceSpontaneous development of spindle cell sarcoma in mdx micePoster presentation
- 第60回日本小児血液・がん学会学術集会, Nov. 2018, English, 京都市, Domestic conferenceQuantitation of minimal residual disease by droplet digital PCR in high-risk neuroblastoma patientsOral presentation
- The 50th Congress of the International Society of Paediatric Oncology, Nov. 2018, English, 京都市, International conferenceEXPRESSION PROFILES OF SEVEN MINIMAL RESIDUAL DISEASE MARKERS IN BONE MARROW AND PERIPHERAL BLOOD OF PATIENTS WITH HIGH‑RISK NEUROBLASTOMAPoster presentation
- 第80回日本血液学会学術集会, Oct. 2018, Japanese, 大阪市, Domestic conferenceダウン症候群に伴う小児骨髄性白血病の第二再発にアザシチジンが奏功した一例Poster presentation
- 第77回日本癌学会学術総会, Sep. 2018, English, 大阪市, Domestic conferenceDetection of minimal residual disease in high-risk neuroblastoma patients by digital PCROral presentation
- 第60回日本小児神経学会学術集会, Jun. 2018, English, 千葉市, Domestic conferenceWhen does SMN shortage become evident in spinal muscular atrophy patients?Oral presentation
- 第60回日本小児神経学会学術集会, Jun. 2018, English, 千葉市, Domestic conferenceIs congenital cytomegalovirus infection associated with autism spectrum disorder?Oral presentation
- 第274回日本小児科学会兵庫県地方会, May 2018, Japanese, 神戸市, Domestic conferenceMycobacterium chelonaeによる菌血症を発症したダウン症候群に伴う骨髄性白血病の女児例Oral presentation
- Neuroblastoma Research Association 2018, May 2018, English, San Francisco, International conferenceElevated minimal residual disease marker expression in peripheral blood of three high‑risk neuroblastoma casesPoster presentation
- 第121回日本小児科学会学術集会, Apr. 2018, Japanese, 福岡市, Domestic conference日齢38で発症した後天性サイトメガロウイルス感染による血小板減Poster presentation
- 第121回日本小児科学会学術集会, Apr. 2018, Japanese, 福岡市, Domestic conferenceプロプラノロールシロップで治療した乳児血管腫9例の検討Poster presentation
- 第121回日本小児科学会学術集会, Apr. 2018, Japanese, 福岡市, Domestic conference5年以上のRomiplostim投与後に無治療経過観察に至った幼児慢性免疫血小板減少性紫斑病Oral presentation
- AACR Pediatric Cancer Research: From Basic Science to the Clinic, Dec. 2017, English, American association for cancer research, Atlanta, USA, International conferenceA pediatric ETV6-ABL1-positive acute lymphoblastic leukemia case with ETV6-ABL1-indepent acquired resistance to tyrosine kinase inhibitorPoster presentation
- 日本人類遺伝学会 第62回大会, Nov. 2017, Japanese, 日本人類遺伝学会, 神戸, Domestic conferenceGenotype-phenotype correlation in patients with chromosome 13q partial deletionPoster presentation
- 第59回日本小児血液・がん学会学術集会, Nov. 2017, Japanese, 京都市, Domestic conference単一施設における骨肉腫に対する大量化学療法についての後方視的検討Oral presentation
- 第79回日本血液学会学術集会, Oct. 2017, Japanese, 日本血液学会, 東京, Domestic conferencePediatric acute lymphoblastic leukemia with ETV6-ABL1Poster presentation
- 第272回日本小児科学会兵庫県地方会, Sep. 2017, Japanese, 日本小児科学会, 姫路, Domestic conference発達の遅れを機にアレイCGHで診断したSmith-Magenis症候群の1例Oral presentation
- 第272回日本小児科学会兵庫県地方会, Sep. 2017, Japanese, 日本小児科学会, 姫路, Domestic conference日齢38で発症した血小板減少症の男児Oral presentation
- 第53回日本周産期・新生児医学会学術集会, Jul. 2017, Japanese, 日本周産期・新生児医学会, 横浜, Domestic conferenceUBアナライザーとニホンウナギ由来の蛍光タンパク質（UnaG）を用いた新たなアンバウンドビリルビン測定法の開発Public symposium
- 第53回日本周産期・新生児医学会学術集会, Jul. 2017, Japanese, 日本周産期・新生児医学会, 横浜, Domestic conferenceSGA児の胎盤における胎盤形成遺伝子Retrotransposon-like1(Rtl1)メチル化異常の検討Public symposium
- 10th International Conference on HHV-6&7, Jul. 2017, English, 10th International Conference on HHV-6&7, ベルリン, ドイツ, International conferenceExpression of CD134(OX40) on T cells is a trigger of human herpesvirus 6B reactivation and replication after hematopoietic cell transplantationPoster presentation
- 第117回日本小児精神神経学会, Jun. 2017, Japanese, 日本小児神経学会, 東京, Domestic conference乳幼児期における発達障害特性と感覚特性との関連についての検討Oral presentation
- 第59回日本小児神経学会学術集会, Jun. 2017, Japanese, 日本小児神経学会, 大阪, Domestic conference乳幼児期における発達障害児の感覚特性についての検討～自閉症および知的障害特性との関連～Oral presentation
- 第31回ヘルペスウイルス研究会, Jun. 2017, Japanese, 第31回ヘルペスウイルス研究会, 松江, Domestic conference造血幹細胞移植後のT細胞におけるCD134(OX40）の発現が、移植後HHV-6B再活性化および増殖の誘因となるOral presentation
- 第16回日本再生医療学会総会, May 2017, Japanese, 日本再生医療学会, 仙台, Domestic conference臍帯組織中に存在する非腫瘍性多能性幹細胞Muse細胞の機能解析Oral presentation
- 270回 日本小児科学会兵庫県地方会, May 2017, Japanese, 日本小児科学会, 尼崎, Domestic conference乳幼児感覚プロファイルを用いた発達障害児の感覚特性に関する検討Oral presentation
- 第16回日本再生医療学会総会, May 2017, Japanese, 日本再生医療学会, 仙台, Domestic conference早産児の臍帯由来間葉系幹細胞の増殖におけるWNTシグナル経路の役割Oral presentation
- 第30回近畿小児科学会, May 2017, Japanese, 日本小児科学会, 大阪, Domestic conference前処置なしで造血幹細胞移植を行い混合キメラで推移しているX連鎖重症複合免疫不全症Oral presentation
- 第16回日本再生医療学会総会, May 2017, Japanese, 日本再生医療学会, 仙台, Domestic conferenceブレオマイシン誘発肺障害モデルラットにおける臍帯由来間葉系幹細胞の効果Oral presentation
- The 2nd Congress of joint European Neonatal Society, May 2017, English, MCA Scientific Events, Venice, Italy, International conferenceA new evaluation index for left-right unequal ventriculomegaly in very low birth weight infants: Association with walking disabilities at 1.6 years of corrected agePoster presentation
- 第61回日本新生児成育医学会, Dec. 2016, Japanese, 日本新生児成育医学会, 大阪, Domestic conference臍帯由来間葉系幹細胞のWntシグナル経路の遺伝子発現は児の在胎週数により異なるOral presentation
- 第58回日本小児血液・がん学会学術集会, Dec. 2016, Japanese, 日本小児血液・がん学会, 東京, Domestic conferencePET-MRI所見に基づく肺・下鼻甲介・腎生検が原疾患の診断に有効であった二次性血球貪食リンパ組織球症の1例Poster presentation
- 第58回日本小児血液・がん学会学術集会, Dec. 2016, Japanese, 日本小児血液・がん学会, 東京, Domestic conferenceG-CSF投与と自家末梢血幹細胞移植を併用しbi-weekly VDC-IE療法を完遂し得た切除不能ユーイング肉腫例Oral presentation
- 第79回日本血液学会学術集会, Oct. 2016, Japanese, 日本血液学会, 横浜, Domestic conferenceReemergence of a translocation t(11;19)(q23;p13.1) in the absence of leukemiaPoster presentation
- 第78回日本血液学会学術集会, Oct. 2016, Japanese, 日本血液学会, 横浜, Domestic conferenceAggressive double-hit leukemia/lymphoma with t(14;18)(q32;q21) and t(8;21)(q24;q11.2)Poster presentation
- 第52回日本周産期・新生児医学会学術集会, Jul. 2016, Japanese, 日本周産期・新生児医学会, 富山, Domestic conference臍帯由来間葉系幹細胞を用いた脊髄性筋萎縮症の病態解析の試みPoster presentation
- 第52回日本周産期・新生児医学会学術集会, Jul. 2016, Japanese, 日本周産期・新生児医学会, 富山, Domestic conference極低出生体重児の正期産相当時頭部MRIにおける側脳室拡大と修正1歳6ヶ月時歩行確立との関連性Oral presentation
- Advances in Neuroblastoma Research Conference 2016, Jun. 2016, English, Advances in Neuroblastoma Research Conference, Cairns, Australia, International conferenceRab6B mediates the progression of neuroblastoma through the interaction with MTMR5Poster presentation
- Advances in Neuroblastoma Research Conference 2016, Jun. 2016, English, Advances in Neuroblastoma Research Conference, Cairns, Australia, International conferenceDENN domain protein DENND2A regulates the progression of neuroblastoma.Poster presentation
- The 119th Annual Meeting of the Japan Pedeatric Society, May 2016, Japanese, Japan Pediatric Society, 札幌, Domestic conferenceMinimal residual disease monitoring in neuroblastoma patients.Oral presentation
- 第119回日本小児科学会学術集会, May 2016, Japanese, 日本小児科学会, 札幌, Domestic conference次世代シークエンサーによる重症心身障害者施設での原因遺伝子解析Oral presentation
- 第15回日本再生医療学会, Mar. 2016, Japanese, 日本再生医療学会, 大阪, Domestic conference臍帯由来間葉系幹細胞における児の在胎週数による転写制御解析Oral presentation
- 第15回日本再生医療学会, Mar. 2016, Japanese, 日本再生医療学会, 大阪, Domestic conference臍帯組織中に存在する多能性幹細胞Muse細胞の自己複製能と多分化能の検証Oral presentation
- The 57th Annual Meeting of the Japanese Society of Pediatric Hematology / Oncology, Nov. 2015, Japanese, The Japanese Society of Pediatric Hematology / Oncology, 甲府市, Domestic conferenceDifferential expression of minimal residual disease markers in peripheral blood and bone marrow samples from high-risk neuroblastoma patientsOral presentation
- 第57回日本小児血液がん学会学術集会, Nov. 2015, Japanese, 日本小児血液がん学会, 山梨, Domestic conference外来化学療法中（ALL B12臨床試験）の中心静脈カテーテルの管理についての検討Poster presentation
- 第42回日本臓器保存生物医学会学術集会, Nov. 2015, Japanese, 日本臓器保存生物医学会, 盛岡, Domestic conference「細胞移植の新たな展開」臍帯組織中に存在する多能性幹細胞Muse細胞の多能性の解析Public symposium
- 第57回日本小児血液がん学会学術集会, Nov. 2015, Japanese, 日本小児血液がん学会, 山梨, Domestic conferenceDifferential expression of minimal residual disease markers in peripheral blood and bone marrow samples from high-risk neuroblastoma patients.Oral presentation
- The 57th Annual Meeting of the Japanese Society of Pediatric Hematology / Oncology, Nov. 2015, Japanese, The Japanese Society of Pediatric Hematology / Oncology, 甲府市, Domestic conferenceDENN domain protein DENND2A mediates the progression of neuroblastomaOral presentation
- 第49回日本小児内分泌学会学術集会, Oct. 2015, Japanese, 日本小児内分泌学会, 東京, Domestic conference神戸市における成長ホルモン治療の適応と想定されるSGA性低身長症の発生頻度Oral presentation
- 第49回日本小児内分泌学会学術集会, Oct. 2015, Japanese, 日本小児内分泌学会, 東京, Domestic conference神戸市におけるLate-preterm児の3歳時低身長の発生頻度Oral presentation
- 第74回日本癌学会学術総会, Oct. 2015, Japanese, 日本癌学会, 名古屋, Domestic conference神経芽腫の発症・進展におけるRab6BとRab28の役割Oral presentation
- 第60回日本新生児成育医学会・学術集会, Oct. 2015, Japanese, 日本新生児成育医学会, 盛岡, Domestic conferenceUBアナライザーとビリルビン誘導蛍光タンパク質（UnaG）を用いた新たなアンバウンドビリルビン測定法の開発Poster presentation
- 第49回日本小児内分泌学会学術集会, Oct. 2015, Japanese, 日本小児内分泌学会, 東京, Domestic conferenceSmall-for-gestational age児の身長のキャッチアップ率：神戸市のpopulation-based縦断研究Poster presentation
- 第49回日本小児内分泌学会学術集会, Oct. 2015, Japanese, 日本小児内分泌学会, 東京, Domestic conferenceSmall-for-gestational ageを伴うlate-preterm児の3歳時低身長発生における出生身長の影響Poster presentation
- 第77回日本血液学会学術集会, Oct. 2015, Japanese, 日本血液学会, 金沢, Domestic conferencePersistent MRSE infection and CV line-related thrombosis in AML patient in neutropenic phase.Oral presentation
- 第74回日本癌学会学術総会, Oct. 2015, Japanese, 日本癌学会, 名古屋, Domestic conferenceDENNドメイン蛋白質DENND2Aによる神経芽腫発の発症・進展の制御機構Oral presentation
- 18th ECCO - 40th ESMO European Cancer Congress, Sep. 2015, English, ECCO/ESMO, Vienna, Austria, International conferenceContribution of cancer-associated fibroblasts and M2-polarized macrophages to neuroblastoma developmentPoster presentation
- 日本解剖学会第61回東北・北海道連合支部学術集会, Aug. 2015, Japanese, 日本解剖学会, 盛岡, Domestic conference臍帯組織中に存在する多能性幹細胞Muse細胞の多能性の解析Oral presentation
- International society for stem cell research annual meeting, Jun. 2015, English, ISSCR, Stockholm, スウェーデン, International conferenceAnalysis of pluripotency in Muse cells derived from human umbilical cord tissue.Poster presentation
- 第265 回 日本小児科学会兵庫県地方会, May 2015, Japanese, 日本小児科学会兵庫県地方会, 神戸, Domestic conference左鎖骨部腫瘤を呈した生後6ヶ月の乳児例Oral presentation
- 第104回日本病理学会総会, Apr. 2015, Japanese, The Japanese Sciety of Pathology, 名古屋, Domestic conference神経芽腫の進展における腫瘍関連炎症細胞と腫瘍関連間質細胞の相互作用の解明Oral presentation
- The 11th Asian Society for Pediatric Research, Apr. 2015, English, Asian Society for Pediatric Research, 大阪, International conferenceTwo high-risk neuroblastoma cases with early detection of tumor relapse/re-growth by consecutive minimal residual disease monitoring.Poster presentation
- The 11th Asian Society for Pediatric Research, Apr. 2015, English, Asian Society for Pediatric Research, 大阪, International conferenceNeuroblastoma minimal residual disease markers are differentially expressed in peripheral blood and bone marrow samples.Poster presentation
- Pediatric Academic Societies Annual Meeting, Apr. 2015, English, Pediatric Academic Societies, San Diego, アメリカ, International conferenceA novel measurement method for serum unconjugated bilirubin concentrations using a bilirubin-inducible fluorescent protein.Poster presentation
- Pediatric Cancer at The Crossroad, Nov. 2013, English, American Association for Cancer Research, San Diego, USA, Purpose: Neuroblastoma is the most common extra-cranial solid tumor that accounts for ~15% of all cancer-related deaths in children. Despite current aggressive therapies, more than half of high-risk neuroblastoma patients have experienced a tumor relapse leading to mostly cancer-related deaths. As in most cancers, recurrent neuroblastoma is primarily driven by chemoresistant ca, International conferenceIdentification of ALDH1A2 as a critical ALDH isoform in neuroblastoma cells with cancer stem cell phenotypePoster presentation
- 第54回日本小児血液・がん学会学術集会, Dec. 2012, Japanese, 日本小児血液・がん学会, 横浜, 【諸言】神経芽腫の予後は全体的には改善したが、進行例の長期予後は未だ不良であり、微小残存病変 (MRD) の正確 な評価が予後改善には不可欠である。今回我々は、MRD 検索の感度向上のため、multiple real-time RT-PCR marker を 用いた MRD 測定系を樹立し、各マーカーの感度、臨床情報との相関について解析を行った。【方法】既報の 11 の MRD マーカー (CHRNA3, CRMP1, DBH, DCX, DDC, GABRB3, GAP43, ISL1, KIF1A, PHOX2B, TH) について、神経芽腫 tumor initiating cells を段階的に希釈して検量線を作成し 、基準値を設定した。倫理委員会の 承認の下、2011 年 2 月から 2012 年 6 月までに当科で経験した, Domestic conference神経芽腫症例におけるmultiple real-time RT-PCR markerを用いた微小残存病変(MRD)解析Oral presentation
- 日本人類遺伝学会 第57回大会, Oct. 2012, English, The Japan Society of Human Genetics, 東京, [Background] Spinal muscular atrophy (SMA) is an autosomal recessive inherited neuromuscular disease that is caused by loss of the survival motor neuron gene, SMN1. Every SMA patient retains SMN2 which encodes the same protein (SMN) as SMN1 does. However, alternative splicing of SMN2 produces much amount of exon 7-lacking (△7) SMN2 transcript and hampers the production of ful, Domestic conferenceSalbutamol modulates SMN2 expression in SMA fibroblastOral presentation
- Advances in Neuroblastoma Research 2012 Conference, Jun. 2012, English, Tronto, Background: Neuroblastoma is characterized by its tumor heterogeneity driven by the differentiation of tumor-initiating cells (TICs) that can be isolated as spheres. Although many diagnostic and/or prognostic biomarkers have been proposed, the response of, International conferenceRab15 alternative splicing correlates with differentiation of neuroblastoma cells.Poster presentation
- Advances in Neuroblastoma Research 2012 Conference, Jun. 2012, English, Tronto, Background: Minimal residual disease (MRD) is derived from tumor-initiating cells (TICs) consisting of only a small population of tumor, and its activation leads to tumor relapse. Because over 50% of high-risk neuroblastoma patients experience tumor relap, International conferenceMinimal residual disease monitoring in neuroblastoma patients by a set of real-time RT-PCR markers.Poster presentation
- The 54th Annual Meeting of the Japanese Sciety of Child Neurology, May 2012, Japanese, The Japanese Society of Child Neurology, 北海道, 【目的】脊髄性筋萎縮症（SMA）は，SMN2遺伝子コピー数が少なければ重症化することが報告されてきたが，SMN2遺伝子コピー数が少ない軽症例も存在する．一方，SMN1/2遺伝子プロモーターの塩基配列はすべて同一であると報告されてはいるが，十分には研究されてこなかった．今回，我々は，SMN2遺伝子コピー数以外の症状修飾因子を明らかにする目的で，SMN2遺伝子コピー数による予測よりも軽症の経過をとった症例について，プロモーター解析を行った．また，SMNI/2遺伝子プロモーター多型性の頻度についても報告する．【症例と方法】症例は2歳男児．1歳6か月時には伝い歩きが可能であった．遺伝子解析の結果，SMN1遺伝子は欠失し，SMN2遺伝子は2コピー存在していた．（1）症例のSMN2遺伝子プロモーター領域のダイレクトシークエンスを行った．（2）SMN1遺伝子欠失群, Domestic conferenceSMN2遺伝子量解析による予測よりも軽症の経過をとった脊髄性筋萎縮症患者に対するプロモーター解析Oral presentation
- 第82回日本衛生学会総会, Mar. 2012, Japanese, 日本衛生学会, 京都, Domestic conferenceMolecular analysis of SMN genes in the patients who were clinically diagnosed as habing SMA.Poster presentation
- 第82回日本衛生学会総会, Mar. 2012, Japanese, 日本衛生学会, 京都, Domestic conferenceA study on anti-cancer effect of tea catechin against neuroblastoma fumor initiating cells.Oral presentation
- 第53回日本小児血液・がん学会学術集会, Nov. 2011, Japanese, 日本小児血液・がん学会, 前橋, Domestic conferenceA case of alveolar rhabdomyosarcoma mimicking acute leukemia of admissionPoster presentation
- 日本小児科学会雑誌, Dec. 2010, JapaneseSMN1遺伝子の片側アレルの欠失を認め、もう一方のアレルに点突然変異を認めた脊髄性筋萎縮症の1例
- 第250回日本小児科学会兵庫県地方会, May 2010, Japanese, 神戸, Domestic conferenceSMN1遺伝子の片側アレルの欠失を認め、もう一方のアレルに点突然変異を認めた脊髄性筋委縮症の1例Oral presentation
- 第81回日本生化学会大会・第31回日本分子生物学会年会合同大会, Dec. 2008, JapaneseRab8/13-JRAB/MICAL-L2 complexes control epithelial apical junctions[Invited]
- 第60回日本細胞生物学会大会, Jun. 2008, Japanese高次細胞機能発現におけるエキソサイトーシス系Rab ファミリー低分子量G蛋白質の作用機構
- 第66回日本癌学会総会, Oct. 2007, JapaneseActinin-4によるJRAB/MICAL-L2の細胞膜局在の制御機構
- 第66回日本癌学会総会, Oct. 2007, Japaneseタイトジャンクションとアドヘレンスジャンクションの形成におけるJRAB/MICAL-L2とRab8/13の役割
- 第59回日本細胞生物学会, May 2007, JapaneseJRAB/MICAL-L2の細胞膜局在におけるactinin-4の役割
- 第59回日本細胞生物学会, May 2007, JapaneseDoc2α modulates secretory lysosome exocytosis in mast cells
- 第65回日本癌学会総会, Sep. 2006, JapaneseRab13結合蛋白質JRABによるタイトジャンクション形成の制御機構
- 第65回日本癌学会総会, Sep. 2006, Japanese上皮細胞の細胞分散(cell scattering)におけるRab13-JRAB系の役割
- 第45回日本生化学会中国・四国支部例会, May 2004, JapaneseRabファミリー低分子量G蛋白質による細胞極性・接着の制御機構
- 第104回日本外科学会定期学術集会, Apr. 2004, JapaneseRabファミリー低分子量G蛋白質Rab13によるタイトジャンクションの接着分子Occludinの小胞輸送制御機構
- 第62回日本癌学会総会, Sep. 2003, JapaneseRabファミリー低分子量G蛋白質によるタイトジャンクション膜蛋白質の小胞輸送の制御機構
- 第56回日本細胞生物学会大会, May 2003, Japanese細胞極性と接着を支える小胞輸送の制御機構：Rabファミリー低分子量G蛋白質の役割
- 日本学術振興会, 科学研究費助成事業 基盤研究(C), 基盤研究(C), 神戸大学, 01 Apr. 2021 - 31 Mar. 2024神経芽腫のがん微小環境制御における間葉系幹細胞の役割に関する研究神経芽腫は、神経堤細胞が交感神経系へ分化する過程で発生する代表的な小児難治性固形がんで、小児がん死亡の約1/6を占めている。特に、高リスク神経芽腫患者の大部分は、一旦は治療に反応して寛解を達成するが、その半数以上が再発して極めて治療困難になり、その長期生存率は未だ50%に達していない。これには、治療後に微小残存病変(MRD)として体内に残存したがん細胞が再活性化し、異なる形質を示すようになることが重要だと考えられる。化学療法や放射線療法は、治療後に亜致死となったがん細胞および微小環境中の間質細胞に細胞老化を誘導(治療誘発細胞老化：TIS)し、エクソソームやサイトカイン等の分泌(細胞老化随伴分泌現象：SASP)を促していることが明らかになり、治療困難のメカニズムを理解するためには、がん細胞の異なる形質の発現を担う分泌因子の同定が必須だと考えられる。これまでに申請者らは、MRDの新規評価法を開発してその動態を明らかにすると共に、神経芽腫細胞と微小環境の主要な構成細胞で分泌活性の高い間葉系幹細胞(MSC)との相互作用を明らかにしてきた。そこで本研究では、MSCのSASPによって分泌される分子を同定し、その機能を明らかにすることを試みる。本年度の研究では、高リスク神経芽腫治療に用いられるシスプラチン(CDDP)やテモゾロミド(TMZ)によるMSCのSASPによって分泌される分子群を同定した。
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe Gakuin University, 01 Apr. 2021 - 31 Mar. 2024Myostatin antisense nucleic acid therapy for rhabdomyosarcoma横紋筋肉腫(Rhabdomyosarcoma:RMS)は、筋原性の高悪性腫瘍で集学的治療により予後は改善した。しかし、治療抵抗群もあり、より効果的な治療法の開発が喫緊の課題である。ミオスタチンは筋肉の増殖を抑制するミオカインで、筋萎縮の治療法としてミオスタチン阻害に関する研究が盛んである。申請者は、ミオスタチン遺伝子(MSTN)のスプライシングを阻害してミオスタチンの産出を抑えるアンチセンス核酸(MSTN-ASO)の開発に成功した。このMSTN-ASOは想定通り筋芽細胞の増殖を促進した。一方で、同じMSTN-ASOがRMS細胞の増殖を阻害することを示唆する結果が得られていた。 今年度は、まず、RMS細胞でMSTN-ASOが増殖を阻害することを明確にした。RMS細胞のASOによる増殖阻害とMSTN発現レベルを比較した結果、MSTN-ASOによる増殖阻害はMSTNが発現しているRMS細胞でみられることが示された。MSTN-ASOによる増殖阻害がみられたRMS細胞ではMSTN-ASOによるMSTN mRNAの産生抑制と、ミオスタチンシグナルの低下が観察された。次にRMS細胞の増殖阻害を細胞周期に注目して検討を行った。細胞周期インジケーターFast-Fucciを利用し細胞周期停止を検証したところ、S期の進行が阻害されていることが示唆された。今後は細胞周期停止に関わる因子の同定とマウスxenograftモデルでの解析を進める。
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), Kobe University, 01 Apr. 2017 - 31 Mar. 2020, Principal investigatorHigh-risk neuroblastoma causes more than 50% relapse and shows therapy resistance. This is mainly due to tumor heterogeneity that is induced by tumor microenvironment and therapy stress and controlled by Rab family small G proteins (Rabs), key regulators of membrane traffic. In the present study, we have identified some members of Rabs, which potentially control the secretion of growth factors and tumor markers in neuroblastoma microenvironment.Competitive research funding
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), 01 Apr. 2016 - 31 Mar. 2019Dystrophin intron retention analysis to identify new targets for Antisense Oligonucleotide mediated RNA modulation in RhabdomyosarcomaDystrophin is regarded as a potential tumor suppressor gene because it was shown to be mutated in a variety of rhabdomyosarcoma and DMD patients sometimes develop some kind of tumors. Therefore, this project was intended to understand the significance of dystrophin as a tumor suppressor in rhabdomyosarcoma. In this project, the applicants showed that dystrophin intron retention was an important factor of rhabdomyosarcoma formation. They next, designed an antisense oligonucleotide to target the removal of this retained intron. Abolishment of the retained intron was successful. In addition, they showed that abolishing intron retention, increased dystrophin production of a carboxyl-terminal dystrophin isoform. Moreover, the cell proliferation of the rhabdomyosarcoma reduced drastically when intron retention was abolished. They equally demonstrated the cell specificity, sensitivity and specie specificity of the antisense oligonucleotide. This is antisense could be potential drug for RMS
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), Grant-in-Aid for Young Scientists (B), Kobe University, 01 Apr. 2016 - 31 Mar. 2018Role of DENND2A for development of neuroblastomaNeuroblastoma is a typical recurrent and refractory tumor in children and its long-term survival remains less than 40%. Neuroblastoma relapse is caused by activation of chemo-resistant cancer stem cells (CSCs). Key regulators of intracellular vesicle trafficking, DENN domain proteins, were implicated in this process. In this study, a member of DENND domain proteins DENND2A and its target Rab family small G protein Rab9B were found to be involved in the development and progression of neuroblastoma.
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), Grant-in-Aid for Young Scientists (B), Kobe University, 01 Apr. 2014 - 31 Mar. 2017Mesenchymal stem cell therapy for neonatal hypoxic ischemic encephalopathyTreatments for neonatal hypoxic ischemic encephalopathy have been limited. Mesenchymal stem cells (MSCs) have significant potential for use in cell therapy for various diseases, through their ability to home to sites of injury, suppress immune responses, and undergo self-renewal and differentiation. The objective of this study is to characterize the gestational age-dependent change in MSCs from fetal tissues and to assess whether the MSCs could be useful for neonatal brain injury due to hypoxia or ischemia. First, we isolated umbilical cord-derived MSCs(UC-MSCs) from infants delivered at 22 to 40 weeks gestational age. UC-MSCs from preterm infants showed high proliferative capacity with increased levels of Wnt signaling pathway gene expression compared with those from term infants. Second, the lung injuries on rat model of bleomycin-induced hypoxemia and pulmonary fibrosis could be reduced by UC-MSCs, regardless of origin’s gestation.
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe University, 01 Apr. 2014 - 31 Mar. 2017, Principal investigatorMore than half of high-risk neuroblastoma patients have experienced tumor relapses and their overall survival rates remain less than 40%. To improve their prognosis, it is necessary to develop a novel therapeutic approach targeting minimal residual disease (MRD) comprised of chemoresistant cancer stem cells (CSCs). In the present study, we have identified a member of Rab family small G proteins (Rabs), which control the development of neuroblastoma CSCs, and its effector protein. We have then characterized their upstream/downstream signaling pathways.Competitive research funding
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), Grant-in-Aid for Young Scientists (B), Kobe University, 01 Apr. 2014 - 31 Mar. 2016Role of DENN domain proteins in the development of neuroblastoma cancer stem cells.More than half of high-risk neuroblastoma patients experience tumor relapses and less than 40% can expect a long-term survival. Intracellular vesicle transport seems to play an important role on the development of neuroblastoma cancer stem cells that are responsible for tumor relapses. In this study, we focused on DENN domain proteins, regulators of intracellular vesicle transport, and revealed that a member of DENN domain proteins is involved in the progression of neuroblastoma.
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe University, 01 Apr. 2013 - 31 Mar. 2016The development of new therapy for neonatal chronic lung disease using mesenchymal stem cells derived from fetal appendages.Chronic lung disease (CLD) of prematurity is a common and serious complication in preterm infants. To date, there is no effective treatment for CLD. Perinatal inflammation and regulation of the immune responses have been reported to play a crucial role in the development of CLD. Mesenchymal stem cells (MSCs) are multipotent stromal cells that can differentiate into a variety of cell types, including osteocytes, adipocytes and chondrocytes. Recently, MSCs have been reported to secrete a variety of factors that promote tissue repair and decrease inflammatory and immune reactions. The objective of study was to develop new therapy for CLD using MSCs isolated from fetal appendages in premature infants. In this study, we successfully established a simple and efficient method isolating MSCs from umbilical cord. Furthermore, to determine the effectiveness of MSCs for CLD, we developed an experimental animal model of intra-tracheal bleomycin-induced lung injury.
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe University, 01 Apr. 2013 - 31 Mar. 2016Spinal muscular atrophy (SMA) is a genetic neuromuscular disorder. SMA is caused by the loss of the SMN1 gene. Although the SMN2 gene also produces SMN protein, it is not enough to compensate for the loss of the SMN1 gene. Salbutamol (adrenergic drug) was reported to improve SMA symptoms. To clarify the mechanism, we treated SMA fibroblasts lacking the SMN1 gene with salbutamol, and analyzed SMN2 mRNA and SMN protein levels. Salbutamol increased SMN protein levels in a dose-dependent manner, although SMN2 mRNA levels were not changed. The salbutamol-induced increase in SMN protein level was blocked by PKA inhibitor and deubiquitinase inhibitor. Immunoprecipitation assay using HeLa cells showed that salbutamol decreased ubiquitinated SMN protein levels, suggesting that salbutamol inhibited ubiquitination. These findings suggest that salbutamol increases SMN protein levels in SMA cells by inhibiting ubiquitin-mediated SMN degradation via activating β2-adrenergic receptor-PKA pathways.Competitive research funding
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe University, 2011 - 2013, Principal investigatorMore than 50% of high-risk neuroblastoma patients have experienced tumor relapses caused by chemoresistant cancer stem cells (CSCs). Consequently, their overall survival rates remain less than 40%. To improve their prognosis, it is essential to understand the molecular mechanism of how CSCs are generated and maintained in neuroblastoma. In the present study, we have focused on the key regulators of membrane traffic, Rab family small G proteins (Rabs), and identified a member of Rabs controlling the development of neuroblastoma CSCs.Competitive research funding
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe University, 2010 - 2012Establishment of treatment strategy for spinal muscular atrophybased on the SMN2gene transcription controlMore than 95 % of spinal muscular atrophy (SMA) patients show homozygous deletion of SMN1. SMN2, a highly homologous gene of SMN1, compensates for the SMN1deletion to some degree; copy number of SMN2is inversely correlated to the clinical severity of SMA. Although the promoter sequences of the two genes are almost identical, c.-318 GCC insertion has been identified as a specific polymorphism to the SMN1 promoter. In this study, we found c.-318 GCC insertion polymorphism in the SMN2 promoter of an SMN1-deleted SMA patient with milder phenotype than expected for low copy number of SMN2. However, transcript amount of SMN2in the white blood cells was smaller than other five SMN1-deleted SMA patients, suggesting that the polymorphism did not increase the transcriptional activity. Besides, reporter gene assay using plasmid constructs with or without c.-318 GCC insertion polymorphism demonstrated that the polymorphism had a slightly negative effect on the transcription efficiency. In conclusion,c.-318 GCC insertion polymorphism in the SMN2 promoter may not be associated with the milder phenotype of the patient, suggesting the presence of non-SMN2-related modifying factors of SMA severity. Our experimental data using plasmid constructs with or without c.-318 GCC insertion polymorphism suggested that in thecase of medical treatment for SMA, it is necessary to change the kind and quantity of the SMN2 -activating medicine by the presence of c.-318 GCC insertion polymorphism.
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe University, 2009 - 2011Neuroblastoma is an aggressive solid tumor that accounts for approximately 15% of all cancer-related deaths in children. More than half of high-risk neuroblastoma patients are estimated to experience relapses and their overall survival rates remain less than 40%. Identification of a new prognostic factor is essential to improve the prognosis of neuroblastoma patients. Aberrant alternative splicing is intimately associated with an increasing number of cancers, and its use as a new diagnostic and/or prognostic biomarker has attracted considerable attention. In the present study, we identified four alternatively spliced isoforms of Rab15, originally isolated as a brain-specific Rab protein. Rab15 isoform balance correlated with differentiation of neuroblastoma tumor-initiating cells(TICs) and Rab15 alternative splicing may serve as a biomarker to discriminate TICs from non-TICs in neuroblastoma.Competitive research funding
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), The University of Tokushima, 2008 - 2010Role of Rab13 binding protein JRAB/MICAL-L2 in regulating cell polarity and adhesionDynamic rearrangement of epithelial cell junctions, tight junction (TJ) and adherens junction (AJ), is essential to maintain the homeostasis of the organism in response to various stimuli. The transport of integral TJ and AJ proteins, claudins, occludins and cadherins, to and/or from the plasma membrane enables this rearrangement. We previously revealed that Rab13 and JRAB/MICAL-L2 (JRAB) mediated the endocytic recycling of occludin. In the present study, we identified Rab8 and actinin-4 as JRAB-binding proteins by using yeast two-hybrid screening. Rab13 and Rab8 competed with each other for the binding to JRAB and functionally associated with JRAB at the plasma membrane and recycling endosome, respectively. Furthermore, Rab13-bound JRAB was targeted to the plasma membrane through its binding to actinin-4. These results suggest that JRAB regulates epithelial cell polarity and adhesion through its binding to Rab13, Rab8 and actinin-4.
- 科学研究費補助金／基盤研究(C), 2010Competitive research funding
- 科学研究費補助金／基盤研究(C), 2009, Principal investigatorCompetitive research funding
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), The University of Tokushima, 2006 - 2007Role of Rab 13 and its effector in establishing cell polarityTight junction (TJ) and adherens junction (AJ) are located at the apical end of the basolateral membrane of polarized epithelial cells. During polarization, epithelial cells first form spot-like primordial AJ at the tips of the initial cell-cell contacts and develop belt-like mature AJ. In parallel with the maturation of AJ, TJ is formed at the apical side of AJ. Whereas epithelial polarization entails a complex interplay between the coordinated TJ and AJ assembly and several fundamental cellular processes, the transport of integral TJ and AJ proteins to and/or from the plasma membrane is essential for the control of TJ and AJ assembly. We previously reported that Rab 13 and a junctional Rab13-binding protein(JRAB)/molecule interacting with CasL-like 2 (MICAL-L2) mediated the endocytic recycling of an integral TJ protein occludin and the formation of functional TJ. In the present study, we examined the role of Rab13 and JRAB/MICAL-L2 in the transport of other integral TJ and AJ proteins claudin-1 and E-cadherin to the plasma membrane. Although knockdown of Rab13 specifically inhibited claudin-1 and occludin but not E-cadherin transport, knockdown of JRAB/MICAL-L2 and expression of its Rab13-binding domain(JRAB/MICAL-L2-C)retarded claudin-1, occludin, and E-cadherin transport. We then identified Rab8 as another JRAB/MICAL-L2-C-binding protein. Knockdown of Rab8 inhibited the Rab13-independent transport of E-cadherin to the plasma membrane. Rab13 and Rab8 competed with each other for the binding to JRAB/MICAL-L2 and functionally associated with JRAB/MICAL-L2 at the plasma membrane and recycling endosome, respectively. These results suggest that the interactions of JRAB/MICAL-L2 with Rab13 and Rab8 play a crucial role in establishing cell polarity.
- 日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, 徳島大学, 2003 - 2007多細胞生物の可塑性を支える小胞輸送の制御機構本研究では、Rabファミリー低分子量G蛋白質を中心とした細胞内小胞輸送の制御分子群に焦点を絞って解析し、多細胞生物の可塑性を支える小胞輸送の制御機構について解析を行った。情報の可塑性については、Rab3の活性制御蛋白質であるRab3GEPに直接結合し、Rab3GAPに間接的に結合するシナプス小胞蛋白質Rabconnection-3についてマウス個体と海馬の初代神経培養細胞を用いた解析を行い、本蛋白質がシナプス形成に関与している可能性を示す結果を得ている。一方、形態の可塑性については、これまでに私どもはタイトジャンクション(TJ)の細胞間接着分子であるoccludinのリサイクリングを制御するRab13-JRAB系を見出しているが、本年度はRab13-JRAB系がTJの今ひとつの接着分子claudinの輸送にも関与することを示した。さらに、JRABはRab13以外にRab8とも結合し、Rab8-JRAB系は、アドヘレンスジャンクションの接着分子E-cadherinの細胞膜への輸送に関与することを明らかにした。一方、これまでにJRABがアクチン細胞骨格系に沿って局在することを示していたが、本年度の本研究で、JRABがアクチン結合蛋白質のひとつであるアクチニン4と直接結合することを示した。また、JRABはアクチニン4と結合することによって細胞膜にリクルートされて接着分子の輸送に機能すること、その結合はRab13により促進されることを示した。このように、本年度の本研究は順調に進展し、当初の目的はほぼ達成できた。
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), The University of Tokushima, 2004 - 2005Role of Rab13 in regulating cell polarity and adhesionTight junctions (TJs) are continuous, circumferential belt-like structures located at the apical end of the intercellular space, where they delineate the boundaries between the apical and basolateral domains of the plasma membrane of epithelial cells. TJs are composed of integral TJ proteins and TJ plaque proteins. While integral TJ proteins including occludin and claudins mediate cell-cell adhesion and create physical intercellular barrier, TJ plaque proteins cluster integral TJ proteins and form an organizing platform for a variety of scaffolding, signaling, and vesicular transport proteins. Rab13 is identified as a TJ plaque protein in epithelial cells and is implicated in the assembly of functional TJs. We previously demonstrated that Rab13 specifically mediated the endocytic recycling of occludin. In the present study, we have identified MICAL-L2 (molecule interacting with CasL-like 2) as a novel Rab13-binding protein and renamed it as JRAB (junctional Rab13-binding protein). Immunoprecipitation and immunofluorescence microscopy showed that JRAB specifically bound to the GTP-bound form of Rab13 via its C-terminus, which contained a coiled-coil domain, and localized at TJs in epithelial MTD-1A cells. Recycling assay demonstrated that a JRAB mutant lacking the Rab13-binding domain (JRAB-N) specifically inhibited the endocytic recycling of occludin, but not transferrin receptor. Ca^<2+>-switch assay further revealed that JRAB-N as well as Rab13 Q67L inhibited the recruitment of occludin to the plasma membrane, the development of transepithelial electrical resistance, and the formation of a paracellular diffusion barrier. JRAB was displaced from TJs upon actin depolymerization in MTD-1A cells and was distributed along stress fibers in fibroblastic NIH3T3 cells. These results suggested that JRAB mediated the endocytic recycling of occludin and the formation of functional TJs by linking Rab13 to actin cytoskeleton.
- 日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, 徳島大学, 2003 - 2004メンブレントラフィックにおけるタイトジャンクション膜タンパク質の選別機構タイトジャンクション(TJ)は、物理的に隣り合う細胞間隙の物質透過を制限し、細胞膜を頂端側と側基底側に分離すると共に、細胞膜上の膜ドメインとしてシグナル伝達分子を集積することによって、細胞極性を形成・維持している。TJを構成する膜タンパク質であるクローディンやオクルーディンは、細胞膜まで小胞輸送されてTJを形成すると考えられる。細胞内小胞輸送の代表的な制御系であるRabファミリー低分子量Gタンパク質(Rab)には60以上のメンバーが同定されており、個々のメンバーが特定の細胞内膜系に局在して小胞輸送および膜ドメインの形成を制御している。これまでに、TJ膜タンパク質と頂端側および側基底側膜タンパク質との選別機構を明らかにする目的で、TJに局在するRab13が、クローディンのゴルジ体から細胞膜への輸送およびオクルーディンのエンドソームから細胞膜へのリサイクリングを特異的に制御することを明らかにしてきた。そこで、本年度の研究では、Rab13の作用機構を明らかにする目的で、Rab13の標的タンパク質の同定を試み、以下の結果を得た。1)酵母Two-hybrid法を用いて、GTP結合型Rab13に特異的に結合する分子としてJRABを同定した。2)JRABは脳、肝臓、腎臓、肺に発現し、マウス乳腺上皮(MTD-1A)細胞のTJに局在した。3)JRABのRab13結合領域欠失変異体(JRAB-N)は、オクルーディンの細胞膜へのリサイクリングを特異的に抑制した。4)JRAB-Nは、カルシウムスイッチしたMTD-1A細胞における機能的TJの形成を抑制した。これらの結果から、Rab13は、標的タンパク質JRABを用いて、TJ膜タンパク質と頂端側および側基底側膜タンパク質との選別を制御していることが示唆された。このように、本年度の研究は予想以上に進展し、当初の目的はほぼ達成できた。
- Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), The University of Tokushima, 2002 - 2003Role of the Rab family small G proteins in cell polarity and adhesionTight junctions (TJs) are points of adhesion between epithelial cells that ensure the development and maintenance of the apical and basolateral plasma membrane (PM) domains. Proteins constituting TJs include the transmembrane proteins mediating cell-cell adhesion and the cytosolic plaque proteins linking to the cytoskeleton and participating in the intracellular signaling. Occludin and claudins constitute the transmembrane proteins in TJs. The Rab family small G proteins, which consist of more than 60 family members in mammalian cells, play a crucial role in etermining the specificity of vesicular transport pathways. Two Rab family members, Rab3B and Rab13, localize to TJs in polarized epithelial cells and cytoplasmic vesicular structures in non-polarized fibroblasts, but their functions are poorly understood. In the present study, we have examined the role of Rab3B and Rab13 in regulating the vesicular transport of TJ transmembrane proteins. In the exocytotic pathways, we found that Rab3B and Rab13 direct th cell-surface transport of a basolateral transmembrane protein, low-density lipoprotein receptor, and claudin-1, respectively. We also found that a pool of occludin was continuously endocytosed and recycled back to the cell-surface in epithelial cells. Our results indicated that Rabl3, but not Rab3B, specifically regulated the endocytic recycling of occludin. Biochemical endocytosis and recycling assays demonstrated that Rab13 directed the recycling of endocytosed occludin, but not its endocytosis. Furthermore, we found that Rab13 did not affect the recycling of transferrin receptor, that was recycled to the basolateral PM domain. We isolated a protein that was specifically interacted with GTP-bound form of Rab13, but not its GDP-bound form. Our results suggested that Rab13 was a crucial regulator for the transport of occludin and claudins to TJs.
- 日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, 徳島大学, 2002 - 2002小胞体における積み荷タンパク質の選別機構?輸送シグナルをプローベとして細胞膜タンパク質は、小胞体において積み荷として選別され、さらにゴルジ体の最もトランス側に位置するトランスゴルジ(TGN)において、異なる輸送小胞へ選別されて細胞膜へ輸送される。これまでに、水疱性口内炎ウィルスの糖タンパク質(VSV-G)を細胞膜タンパク質のモデルとして、小胞体〜ゴルジ体間の小胞輸送はシグナル依存的な選択的輸送であること、および、VSV-Gの小胞体からの輸送シグナル(YxDxEシグナル)は、小胞体から発芽するCOPII輸送小胞へVSV-Gを濃縮していることを明らかにしてきた。そこで、本年度の本研究では、YxDxEシグナルの作用機構を明らかにする目的で、YxDxEシグナルを認識する分子の同定を試み、以下の結果を得た。1)酵母Two-hybrid法を用いて、アダプター複合体AP-3のδサブユニットをYxDxEシグナルに結合する分子として同定した。2)YxDxEシグナルとδサブユニットとの結合は、酸性のアスパラギン酸とグルタミン酸のみならずタイロシンにも依存した。3)δサブユニットのVSV-G結合領域を含む欠失変異体は、VSV-Gの小胞体〜ゴルジ体間輸送には作用せず、ゴルジ体〜細胞膜間輸送を特異的に抑制した。4)δサブユニットを欠失したMochaマウスから調製したfibroblastsでは、VSV-Gの小胞体〜ゴルジ体間輸送は正常で、ゴルジ体〜細胞膜間輸送に障害が認められた。5)VSV-Gは、アダプター複合体AP-3依存的にライソソームへ輸送されるLAMP1を細胞膜へミスターゲッティングさせた。これらの結果から、YxDxEシグナルは、小胞体においてはCOPIIコートを、TGNにおいてはアダプター複合体AP-3をシークエンシャルにそれぞれリクルートして、VSV-Gの細胞膜への輸送を制御していることが示唆された。このように、本年度の研究は予想以上に進展し、当初の目的はほぼ達成できた。