Gene therapy for gastric diseases.

Gene therapy for gastric cancer and gastric ulcer is a rationalized strategy since various genes correlate with these diseases. Since gene expressions in non-target tissues/cells cause side effects, a selective gene delivery system targeted to the stomach and/or cancer must be developed. The route of vector transfer (direct injection, systemic, intraperitoneal, gastric serosal surface and oral administration) is an important issue which can determine efficacy and safety. Strategies for cancer gene therapy can be categorized as suicide gene therapy, growth inhibition and apoptosis induction, immunotherapy, anti-angiogenesis, and others. Combination of the target gene with other genes and/or strategies such as chemotherapy and virotherapy is promising. Candidates for treatment of gastric ulcer are vascular endothelial growth factor, angiopoietin-1, serum response factor, and cationic host defense peptide cathelicidin. In this review, we discuss stomach- and cancer-targeted gene transfer methods and summarize gene therapy trials for gastric cancer and gastric ulcer

KAgoshima Galactic Object survey with Nobeyama 45-metre telescope by Mapping in Ammonia lines (KAGONMA): Discovery of parsec-scale CO depletion in the Canis Major star-forming region

In observational studies of infrared dark clouds, the number of detections of CO freeze-out onto dust grains (CO depletion) at pc-scale is extremely limited, and the conditions for its occurrence are, therefore, still unknown. We report a new object where pc-scale CO depletion is expected. As a part of Kagoshima Galactic Object survey with Nobeyama 45-m telescope by Mapping in Ammonia lines (KAGONMA), we have made mapping observations of NH3 inversion transition lines towards the star-forming region associated with the CMa OB1 including IRAS 07077-1026, IRAS 07081-1028, and PGCC G224.28-0.82. By comparing the spatial distributions of the NH3 (1,1) and C18O (J=1-0), an intensity anti-correlation was found in IRAS 07077-1026 and IRAS 07081-1028 on the ~1 pc scale. Furthermore, we obtained a lower abundance of C18O at least in IRAS 07077-1026 than in the other parts of the star-forming region. After examining high density gas dissipation, photodissociation, and CO depletion, we concluded that the intensity anti-correlation in IRAS 07077-1026 is due to CO depletion. On the other hand, in the vicinity of the centre of PGCC G224.28-0.82, the emission line intensities of both the NH3 (1,1) and C18O (J=1-0) were strongly detected, although the gas temperature and density were similar to IRAS 07077-1026. This indicates that there are situations where C18O (J=1-0) cannot trace dense gas on the pc scale and implies that the conditional differences that C18O (J=1-0) can and cannot trace dense gas are unclear.Comment: 19 pages, 15 figures, 4 tables, accepted for Publications of the Astronomical Society of Japan (PASJ). The version 1 is the Author's Original Version. My accepted manuscript will be publicly available on the arXiv one year after publication in the PAS

Multi log-normal density structure in Cygnus-X molecular clouds: A fitting for N-PDF without power-law

We studied the H$_2$ column density probability distribution function (N-PDF) based on molecular emission lines using the Nobeyama 45-m Cygnus X CO survey data. Using the DENDROGRAM and SCIMES algorithms, we identified 124 molecular clouds in the $^{13}$CO data. From these identified molecular clouds, an N-PDF was constructed for 11 molecular clouds with an extent of more than 0.4 deg$^2$. From the fitting of the N-PDF, we found that the N-PDF could be well-fitted with one or two log-normal distributions. These fitting results provided an alternative density structure for molecular clouds from a conventional picture. We investigated the column density, dense molecular cloud cores, and radio continuum source distributions in each cloud and found that the N-PDF shape was less correlated with the star-forming activity over a whole cloud. Furthermore, we found that the log-normal N-PDF parameters obtained from the fitting showed two impressive features. First, the log-normal distribution at the low-density part had the same mean column density ($\sim$ 10$^{21.5}$ cm$^{-2}$) for almost all the molecular clouds. Second, the width of the log-normal distribution tended to decrease with an increasing mean density of the structures. These correlations suggest that the shape of the N-PDF reflects the relationship between the density and turbulent structure of the whole molecular cloud but is less affected by star-forming activities.Comment: 14 pages, 7 Figures, Accepted in MNRA

Highly stomach-selective gene transfer following gastric serosal surface instillation of naked plasmid DNA in rats.

BACKGROUND: The purpose of this study was to achieve stomach-selective gene transfer in rats by our simple and novel administration method, which is gastric serosal surface instillation of naked plasmid DNA (pDNA). METHODS: Naked pDNA encoding firefly luciferase as a reporter gene was instilled onto the gastric serosal surface in male Wistar rats. As controls, we performed intraperitoneal, intragastric and intravenous administration of naked pDNA. At appropriate time intervals, we measured luciferase activities in the stomach and other tissues. RESULTS: Gene expression in the stomach 6 h after gastric serosal surface instillation of naked pDNA (5 microg) was significantly higher than that after using other administration methods. The present study is the first report on stomach-selective gene transfer following instillation of naked pDNA onto the gastric serosal surface in rats. Also, the gene expression level in the stomach 6 h after gastric serosal surface instillation of naked pDNA was markedly higher than that in other tissues. In a dose-dependent study, the gene expression level was saturated over 5 microg. Gene expression in the stomach was detected 3 h after gastric serosal surface instillation of naked pDNA. The gene expression level peaked 12-24 h after instillation of naked pDNA, then decreased to a level similar to 3 h at 48 h. CONCLUSIONS: Gastric serosal surface in stillation of naked pDNA can be a highly stomach-selective gene transfer method in rats

Improved stomach selectivity of gene expression following microinstillation of plasmid DNA onto the gastric serosal surface in mice

Stomach-selective gene transfer is a promising approach as a therapeutic strategy for refractory gastric diseases. In this study, we improved the stomach selectivity of gene expression following microinstillation of naked plasmid DNA (pDNA) onto the gastric serosal surface in mice. pDNA encoding firefly luciferase was used as a reporter gene. It was confirmed that the gene expression level in the stomach 6 h after gastric serosal surface microinstillation of pDNA was significantlyhigher than after intragastric, intraperitoneal and intravenous administration. Regarding selectivity ofgene expression, the gene expression level in the stomach after gastric serosal surfacemicroinstillation of 1 μg/1 μL (dose/volume) pDNA was 5.7 times higher than that in the spleen. In our previous study (30 μg/30 μL), the expression level in the stomach was 2.7 times higher than that in the spleen; therefore, the selectivity was 2.1 times higher in this study. When we investigated gene expression at various pDNA solution concentrations, the ratio of the gene expression level in the stomach to that in the spleen was the highest as 1 μg/1 μL of pDNA, which was considered the optimal concentration. Information in this study is useful for further development of target organ-selective gene delivery systems

The whole blood transcriptional regulation landscape in 465 COVID-19 infected samples from Japan COVID-19 Task Force

「コロナ制圧タスクフォース」COVID-19患者由来の血液細胞における遺伝子発現の網羅的解析 --重症度に応じた遺伝子発現の変化には、ヒトゲノム配列の個人差が影響する--. 京都大学プレスリリース. 2022-08-23.Coronavirus disease 2019 (COVID-19) is a recently-emerged infectious disease that has caused millions of deaths, where comprehensive understanding of disease mechanisms is still unestablished. In particular, studies of gene expression dynamics and regulation landscape in COVID-19 infected individuals are limited. Here, we report on a thorough analysis of whole blood RNA-seq data from 465 genotyped samples from the Japan COVID-19 Task Force, including 359 severe and 106 non-severe COVID-19 cases. We discover 1169 putative causal expression quantitative trait loci (eQTLs) including 34 possible colocalizations with biobank fine-mapping results of hematopoietic traits in a Japanese population, 1549 putative causal splice QTLs (sQTLs; e.g. two independent sQTLs at TOR1AIP1), as well as biologically interpretable trans-eQTL examples (e.g., REST and STING1), all fine-mapped at single variant resolution. We perform differential gene expression analysis to elucidate 198 genes with increased expression in severe COVID-19 cases and enriched for innate immune-related functions. Finally, we evaluate the limited but non-zero effect of COVID-19 phenotype on eQTL discovery, and highlight the presence of COVID-19 severity-interaction eQTLs (ieQTLs; e.g., CLEC4C and MYBL2). Our study provides a comprehensive catalog of whole blood regulatory variants in Japanese, as well as a reference for transcriptional landscapes in response to COVID-19 infection

DOCK2 is involved in the host genetics and biology of severe COVID-19

「コロナ制圧タスクフォース」COVID-19疾患感受性遺伝子DOCK2の重症化機序を解明 --アジア最大のバイオレポジトリーでCOVID-19の治療標的を発見--. 京都大学プレスリリース. 2022-08-10.Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge. Here we conducted a genome-wide association study (GWAS) involving 2, 393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3, 289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2 associated with the risk allele in these younger patients. DOCK2 expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n = 61 individuals) identified cell-type-specific downregulation of DOCK2 and a COVID-19-specific decreasing effect of the risk allele on DOCK2 expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target

Overscreening Induced by Ionic Adsorption at the Ionic Liquid/Electrode Interface Detected Using Neutron Reflectometry with a Rational Material Design

Neutron reflectometry (NR) has been utilized to study the electric double layer (EDL) of ionic liquids (ILs), however, further improvement of the sensitivity toward interfacial structure would be desirable. We recently proposed two ways to improve the NR sensitivity toward the EDL structure at the IL/electrode interface (J. Phys. Chem. C, 123 (2019) 9223). First, as the electrode, a thin film of metal (Nb) was used with the scattering length density (SLD) and thickness controlled to sensitively analyze the potential dependent EDL structure. Second, the IL cation and anion were chosen so that they have large size and large SLD difference, both of which also increase the sensitivity. In the present study, we have further explored this rational material design for the sensitivity enhancement, by changing the film metal from Nb to Bi whose SLD is closer to those for two bulk materials: Si and the IL used, trihexyltetradecylphosphonium bis(nonafluorobutanesulfonyl)amide. We successfully observed not only the first ionic layer in the EDL but also the overlayers, revealing that the IL cation is specifically adsorbed on the electrode and that the cation-rich first layer induces overscreening in the overlayers up to the third ionic layer

Human Chorionic Villous Differentiation and Placental Development

In humans, the placenta provides the only fetomaternal connection and is essential for establishing a pregnancy as well as fetal well-being. Additionally, it allows maternal physiological adaptation and embryonic immunological acceptance, support, and nutrition. The placenta is derived from extra-embryonic tissues that develop rapidly and dynamically in the first weeks of pregnancy. It is primarily composed of trophoblasts that differentiate into villi, stromal cells, macrophages, and fetal endothelial cells (FEC). Placental differentiation may be closely related to perinatal diseases, including fetal growth retardation (FGR) and hypertensive disorders of pregnancy (HDP), and miscarriage. There are limited findings regarding human chorionic villous differentiation and placental development because conducting in vivo studies is extremely difficult. Placental tissue varies widely among species. Thus, experimental animal findings are difficult to apply to humans. Early villous differentiation is difficult to study due to the small tissue size; however, a detailed analysis can potentially elucidate perinatal disease causes or help develop novel therapies. Artificial induction of early villous differentiation using human embryonic stem (ES) cells/induced pluripotent stem (iPS) cells was attempted, producing normally differentiated villi that can be used for interventional/invasive research. Here, we summarized and correlated early villous differentiation findings and discussed clinical diseases