B cell-derived GABA elicits IL-10⁺ macrophages to limit anti-tumour immunity

GABAを標的とする抗腫瘍免疫機構 --代謝産物を介した免疫細胞間制御の一端を解明--. 京都大学プレスリリース. 2021-11-10.Small, soluble metabolites not only are essential intermediates in intracellular biochemical processes, but can also influence neighbouring cells when released into the extracellular milieu1-3. Here we identify the metabolite and neurotransmitter GABA as a candidate signalling molecule synthesized and secreted by activated B cells and plasma cells. We show that B cell-derived GABA promotes monocyte differentiation into anti-inflammatory macrophages that secrete interleukin-10 and inhibit CD8⁺ T cell killer function. In mice, B cell deficiency or B cell-specific inactivation of the GABA-generating enzyme GAD67 enhances anti-tumour responses. Our study reveals that, in addition to cytokines and membrane proteins, small metabolites derived from B-lineage cells have immunoregulatory functions, which may be pharmaceutical targets allowing fine-tuning of immune responses

Predictive model of bosentan-induced liver toxicity in Japanese patients with pulmonary arterial hypertension

Bosentan, an endothelin receptor antagonist, has been widely used as a first-line medication for the treatment of pulmonary arterial hypertension (PAH). It has been shown to improve symptoms of hypertension, exercise capacity, and hemodynamics and prolong time to clinical worsening. However, liver dysfunction is a major side effect of bosentan treatment that could hamper the optimal management of patients with PAH. Previously, we demonstrated, using drug metabolism enzymes and transporters analysis, that the carbohydrate sulfotransferase 3 (CHST3) and CHST13 alleles are significantly more frequent in patients with elevated aminotransferases during therapy with bosentan than they are in patients without liver toxicity. In addition, we constructed a pharmacogenomics model to predict bosentan-induced liver injury in patients with PAH using two single-nucleotide polymorphisms and two nongenetic factors. The purpose of the present study was to externally validate the predictive model of bosentan-induced liver toxicity in Japanese patients. We evaluated five cases of patients treated with bosentan, and one presented with liver dysfunction. We applied mutation alleles of CHST3 and CHST13, serum creatinine, and age to our model to predict liver dysfunction. The sensitivity and specificity were calculated as 100% and 50%, respectively. Considering that PAH is a rare disease, multicenter collaboration would be necessary to validate our model.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Current Status of R&D and Device Manufacturing for ITER Procurement by Japan

Several key components of ITER – such as toroidal filed (TF) coils, conductor for center solenoid (CS) coils, components of neutral beam (NB) system, electron cyclotron heating system, divertor, detritiation system, blanket remote handling system, plasma diagnostic systems, and test blanket system – have been designed and manufactured by JADA (Japan Domestic Agency) for ITER project. The recent highlights of progress and achievements by JADA will be reported.JADA procures all TF coil structures (19 sets), and a TF coil structure comprises inboard and outboard structure. The 4th TF coil structure has already been delivered to EU. Regarding the inboard structure, 10 sets of manufacturing have been completed, and 9 sets of welding assembly processes are ongoing. Regarding the outboard structure, 8 sets of manufacturing have been completed, and 11 sets of welding assembly processes are ongoing.JADA assembles 9 TF coils, and an assembly process of the 1st and 2nd TF coil is carrying out. Regarding the 1st TF coil, case closure welding and gap-filling impregnation have been completed, and then its final machining has started. Regarding the 2nd TF coil, the case closure welding has been completed, and the gap-filling impregnation will be started soon. The Final Acceptance Test (FAT) of 1st and 2nd TF coils will be completed by the end of 2019 and March 2020, respectively. JADA has contributed to the construction of the NB test facility (NBTF) at Padua in Italy, which has the identical design to the ITER NB system. Components for high voltage (HV) power supply – such as HV bussing, 1 MV transmission line, DC generators, and the test power supply – were fabricated and were delivered to Padua. The first integration test between components procured by EU and Japan has been completed in July 2019. DC 1.2 MV for 1 hour was applied to the high voltage deck 1 (HVD1, EU) and to the 1 MV insulating transformer (JA) through the 1 MV transmission line (JA). In September 2019, all components from the terminal of JA component to the Beam Source Vessel (EU) were finally connected by installing HV busing as the 1MV feedthrough to beam source. The FAT will be started in 2020.JADA procures 8 gyrotrons for heating and current drive. The FATs have been passed for 1st and 2nd gyrotron. Achieved parameters on CW operation were output power 1MW for 300 s with 50% of efficiency, 95 % of repeatability. 5 kHz power modulation for 60 seconds is also achieved. Regarding 3rd and 4th gyrotron, manufacturing has been completed, and conditioning of the 3rd gyrotron has started. Manufacture of anode/body power supplies for 4 gyrotrons also have been completed.As stated above, procurement activities carried out by JADA make steady progress in accordance with agreed schedule.12th Asia Plasma and Fusion Association Conferenc

Mesenchymal glioblastoma-induced mature de-novo vessel formation of vascular endothelial cells in a microfluidic device.

High vascularization is a biological characteristic of glioblastoma (GBM); however, an in-vitro experimental model to verify the mechanism and physiological role of vasculogenesis in GBM is not well-established. Recently, we established a self-organizing vasculogenic model using human umbilical vein endothelial cells (HUVECs) co-cultivated with human lung fibroblasts (hLFs). Here, we exploited this system to establish a realistic model of vasculogenesis in GBM. We developed two polydimethylsiloxane (PDMS) devices, a doughnut-hole dish and a 5-lane microfluidic device to observe the contact-independent effects of glioblastoma cells on HUVECs. We tested five patient-derived and five widely used GBM cell lines. Confocal fluorescence microscopy was used to observe the morphological changes in Red Fluorescent Protein (RFP)-HUVECs and fluorescein isothiocyanate (FITC)-dextran perfusion. The genetic and expression properties of GBM cell lines were analyzed. The doughnut-hole dish assay revealed KNS1451 as the only cells to induce HUVEC transformation to vessel-like structures, similar to hLFs. The 5-lane device assay demonstrated that KNS1451 promoted the formation of a vascular network that was fully perfused, revealing the functioning luminal construction. Microarray analysis revealed that KNS1451 is a mesenchymal subtype of GBM. Using a patient-derived mesenchymal GBM cell line, mature de-novo vessel formation could be induced in HUVECs by contact-independent co-culture with GBM in a microfluidic device. These results support the development of a novel in vitro research model and provide novel insights in the neovasculogenic mechanism of GBM and may potentially facilitate the future detection of unknown molecular targets

Base-resolution methylomes of gliomas bearing histone H3.3 mutations reveal a G34 mutant-specific signature shared with bone tumors.

Two recurrent mutations, K27M and G34R/V, in H3F3A, encoding non-canonical histone H3.3, are reported in pediatric and young adult gliomas, whereas G34W mutation is prevalent in bone tumors. In contrast to K27M mutation, it remains elusive how G34 mutations affect the epigenome. Here we performed whole-genome bisulfite sequencing of four G34R-mutated gliomas and the G34V-mutated glioma cell line KNS-42 for comparison with gliomas harboring K27M and no mutations in H3F3A and with G34W-mutated bone tumors. G34R-mutated gliomas exhibited lower global methylation levels, similar CpG island (CGI) methylation levels, and compromised hypermethylation of telomere-proximal CGIs, compared to the other two glioma subgroups. Hypermethylated regions specific to G34R-mutated gliomas were enriched for CGIs, including those of OLIG1, OLIG2, and canonical histone genes in the HIST1 cluster. They were notably hypermethylated in osteosarcomas with, but not without, G34W mutation. Independent component analysis revealed that G34 mutation-specific components shared a significant similarity between glioma and osteosarcoma, suggesting that G34 mutations exert characteristic methylomic effects regardless of the tumor tissue-of-origin. CRISPR/Cas9-mediated disruption of G34V-allele in KNS-42 cells led to demethylation of a subset of CGIs hypermethylated in G34R-mutated gliomas. These findings will provide a basis for elucidating epigenomic roles of G34 oncohistone in tumorigenesis