患者由来iPS細胞を用いたチェディアック・東症候群のミエロイド細胞における病態再現

京都大学新制・論文博士博士(医学)乙第13569号論医博第2295号京都大学大学院医学研究科医学専攻(主査)教授 濵﨑 洋子, 教授 生田 宏一, 教授 滝田 順子学位規則第4条第2項該当Doctor of Medical ScienceKyoto UniversityDFA

Novel Heterogenous CHS1 Mutations Identified in Five Japanese Patients with Chediak-Higashi Syndrome

Chediak-Higashi syndrome (CHS) is a rare, autosomal recessive disorder characterized by oculocutaneous albinism, recurrent bacterial infections and progressive neurological dysfunction. We demonstrate novel heterogenous mutations of CHS1, the responsive gene of CHS, identified in five Japanese patients with CHS. Patients 1, 2, and 3 were siblings, and they had albinism of the skin and hair. They all had a heterogenous two-base deletion (c.5541-5542 del AA, p.Q1847fsX1850) in exon 18. Patient 4 had a heterogenous single-base insertion (c.3944-3945 ins C, p.T1315fsX1331) in exon 10. The patient exhibited severe early-onset phenotype and suffered from hemophagocytic lymphohistiocytosis. Patient 5 had two heterogenous nonsense mutations; c.7982C>G, p.S2661X in exon 30 and c.8281A>T, p.R2761X in exon 31. The patient suffered from infections in childhood and had visual disturbance and albinism of the skin and hair. The CHS1 mutations described here have not been reported previously

ORAI1 Genetic Polymorphisms Associated with the Susceptibility of Atopic Dermatitis in Japanese and Taiwanese Populations

Atopic dermatitis is a chronic inflammatory skin disease. Multiple genetic and environmental factors are thought to be responsible for susceptibility to AD. In this study, we collected 2,478 DNA samples including 209 AD patients and 729 control subjects from Taiwanese population and 513 AD patients and 1027 control subject from Japanese population for sequencing and genotyping ORAI1. A total of 14 genetic variants including 3 novel single-nucleotide polymorphisms (SNPs) in the ORAI1 gene were identified. Our results indicated that a non-synonymous SNP (rs3741596, Ser218Gly) associated with the susceptibility of AD in the Japanese population but not in the Taiwanese population. However, there is another SNP of ORAI1 (rs3741595) associated with the risk of AD in the Taiwanese population but not in the Japanese population. Taken together, our results indicated that genetic polymorphisms of ORAI1 are very likely to be involved in the susceptibility of AD

Radio-copper-labeled Cu-ATSM: an indicator of quiescent but clonogenic cells under mild hypoxia in a Lewis lung carcinoma model

The purpose of this study is to reveal characteristics of 64Cu-labeled diacetyl-bis(N4-methylthiosemicarbazone) ([64Cu]Cu-ATSM) duringcell proliferation and hypoxia by autoradiography imaging and immunohistochemical staining.Methods: The intratumoral distributions of [64Cu]Cu-ATSM and [18F]-2-fluoro-2-deoxy-D-gloucose ([18F]FDG) in mice implanted withLewis lung carcinoma (LLC1) tumor cells according to dual autoradiography were compared with the immunohistochemical staining patternsof proliferating markers [Ki-67 and 5-bromo-2\u27-deoxyuridine (BrdU)] and a hypoxic marker (pimonidazole). A clonogenic assay wasperformed using the cells of LLC1 tumor-implanted mice, and it was compared with the distribution of [64Cu]Cu-ATSM.Results: [64Cu]Cu-ATSM mainly accumulated at the edge of tumors, whereas [18F]FDG was distributed inside the tumor and inside the[64Cu]Cu-ATSM accumulation. The number of Ki-67-positive cells/area tended to increase with [18F]FDG accumulation and decrease with[64Cu]Cu-ATSM accumulation. On the other hand, the number of BrdU-positive cells/area was negatively correlated with [18F]FDGaccumulation and positively correlated with [64Cu]Cu-ATSM accumulation. High [64Cu]Cu-ATSM accumulation was found outside thehigh-[18F]FDG-accumulation and pimonidazole-positive regions. Colony formation ability was significantly higher in the tumor cellsobtained from high-[64Cu]Cu-ATSM-accumulation regions than the cells from the intermediate- and the low-accumulation regions.Conclusion: [64Cu]Cu-ATSM accumulation regions in tumor cells indicate quiescent but clonogenic tumor cells under mild hypoxia. [64Cu]Cu-ATSM could play an important role in planning appropriate tumor radiotherapy

Effects of placental transfusion in extremely low birthweight infants: Meta-analysis of long- and short-term outcomes

Background Risks and benefits of increasing placental transfusion in extremely preterm infants (extremely low birthweight [ELBW], \u3c1000 g) are ill defined. We performed a meta-analysis to compare long- and short-term outcomes of ELBW infants in trials of enhanced placental transfusion regimens. Study Design and Methods We conducted a meta-analysis of randomized controlled trials (RCTs) of delayed umbilical cord clamping or umbilical cord milking in compliance with PRISMA and Cochrane Collaborative guidelines for systematic reviews. We searched multiple databases for medical literature up to December 2012. Inclusion criteria were preterm infants less than 30 weeks and less than 1000 g birthweight, randomly assigned to enhanced placental transfusion (either delayed cord clamping or cord milking) versus immediate cord clamping. The primary outcome was standardized neurodevelopmental outcome at 18 to 24 months corrected age using a standardized scale. Several short-term outcomes were also evaluated as secondary variables. Results We found 19 studies of which 10 studies could be included (n = 199). Three reported neurodevelopmental outcomes, none of which showed significant rates of disability. Two reported these at 18 to 24 months (n = 42) but used different scales preventing pooling. Short-term benefits of enhanced placental strategies included better blood pressure and hemoglobin on admission, along with reduced blood transfusions, a trend to reduced intraventricular hemorrhage, and episodes of late-onset sepsis. Conclusions Strategies to enhance placental transfusion may improve short-term outcomes of ELBW infants. However, paucity of data on neurodevelopmental outcomes and safety concerns tempers enthusiasm for these interventions. Appropriately designed RCTs to assess short-term and long-term outcomes are needed in ELBW infants. © 2013 American Association of Blood Banks

Assessment of the Tumor Redox Status in Head and Neck Cancer by 62Cu-ATSM PET.

PURPOSE:Tumor redox is an important factor for cancer progression, resistance to treatments, and a poor prognosis. The aim of the present study was to define tumor redox (over-reduction) using 62Cu-diacetyl-bis(N4-methylthiosemicarbazone) (62Cu-ATSM) PET and compare its prognostic potential in head and neck cancer (HNC) with that of 2-deoxy-2-[18F]fluoro-D-glucose (18F-FDG). METHODS:Thirty HNC patients (stage II-IV) underwent pretreatment 62Cu-ATSM and 18F-FDG PET scans. Maximum standardized uptake values (SUVATSM and SUVFDG) and tumor-to-muscle activity concentration ratios (TMRATSM and TMRFDG) were measured. Reductive-tumor-volume (RTV) was then determined at four thresholds (40%, 50%, 60%, and 70% SUVATSM), and total-lesion-reduction (TLR) was calculated as the product of the mean SUV and RTV for 62Cu-ATSM. In 18F-FDG, metabolic-tumor-volume (MTV) and total-lesion-glycolysis (TLG) were obtained at a threshold of 40%. A ROC analysis was performed to determine % thresholds for RTV and TLR showing the best predictive performance, and these were then used to determine the optimal cut-off values to stratify patients for each parameter. Progression-free-survival (PFS) and cause-specific-survival (CSS) were evaluated by the Kaplan-Meier method. RESULTS:The means ± standard deviations of PFS and CSS periods were 16.4±13.4 and 19.2±12.4 months, respectively. A ROC analysis determined that the 70% SUVATSM threshold for RTV and TLR was the best for predicting disease progression and cancer death. Optimal cut-offs for each index were SUVATSM = 3.6, SUVFDG = 7.9, TMRATSM = 3.2, TMRFDG = 5.6, RTV = 2.9, MTV = 8.1, TLR = 14.0, and TLG = 36.5. When the cut-offs for TMRATSM and TLR were set as described above in 62Cu-ATSM PET, patients with higher TMRATSM (p = 0.03) and greater TLR (p = 0.02) showed significantly worse PFS, while patients with greater TLR had significantly worse CSS (p = 0.02). Only MTV in 18F-FDG PET predicted differences in PSF and CSS (p = 0.03 and p = 0.03, respectively). CONCLUSION:Tumor redox parameters measured by 62Cu-ATSM PET may be determinants of HNC patient outcomes and help define optimal patient-specific treatments

Kaplan-Meier curves of progression-free survival (PFS) for ⁶²Cu-ATSM PET (a) and ¹⁸F-FDG PET (b) in patients with HNC.

<p>Two groups of high (dotted lines) and low (solid lines) tracer accumulation were determined by each cut-off value of the tumor-to-muscle ratios (TMR_ATSM and TMR_FDG). TMR_ATSM, one of the intensity-based redox parameters, showed a significant difference in PFS between two groups (<i>p</i> = 0.03), whereas TMR_FDG, one of the intensity-based metabolic parameters, did not (<i>p</i> = 0.15). The three-year PFS rate was 74% for patients with lower accumulation tumors (TMR_ATSM ≤ 3.2) and 29% for those with over-reductive tumors (TMR_ATSM > 3.2).</p

Patient Characteristics.

<p>Patient Characteristics.</p