Targeting Glycosylation Pathways and the Cell Cycle: Sugar-Dependent Activity of Butyrate-Carbohydrate Cancer Prodrugs

SummaryShort-chain fatty acid (SCFA)-carbohydrate hybrid molecules that target both histone deacetylation and glycosylation pathways to achieve sugar-dependent activity against cancer cells are described in this article. Specifically, n-butyrate esters of N-acetyl-d-mannosamine (But4ManNAc, 1) induced apoptosis, whereas corresponding N-acetyl-d-glucosamine (But4GlcNAc, 2), d-mannose (But5Man, 3), or glycerol (tributryin, 4) derivatives only provided transient cell cycle arrest. Western blots, reporter gene assays, and cell cycle analysis established that n-butyrate, when delivered to cells via any carbohydrate scaffold, functioned as a histone deacetylase inhibitor (HDACi), upregulated p21WAF1/Cip1 expression, and inhibited proliferation. However, only 1, a compound that primed sialic acid biosynthesis and modulated the expression of a different set of genes compared to 3, ultimately killed the cells. These results demonstrate that the biological activity of butyrate can be tuned by sugars to improve its anticancer properties

Common Gene Therapy Viral Vectors Do Not Efficiently Penetrate Sputum from Cystic Fibrosis Patients

Norwalk virus and human papilloma virus, two viruses that infect humans at mucosal surfaces, have been found capable of rapidly penetrating human mucus secretions. Viral vectors for gene therapy of Cystic Fibrosis (CF) must similarly penetrate purulent lung airway mucus (sputum) to deliver DNA to airway epithelial cells. However, surprisingly little is known about the rates at which gene delivery vehicles penetrate sputum, including viral vectors used in clinical trials for CF gene therapy. We find that sputum spontaneously expectorated by CF patients efficiently traps two viral vectors commonly used in CF gene therapy trials, adenovirus (d∼80 nm) and adeno-associated virus (AAV serotype 5; d∼20 nm), leading to average effective diffusivities that are ∼3,000-fold and 12,000-fold slower than their theoretical speeds in water, respectively. Both viral vectors are slowed by adhesion, as engineered muco-inert nanoparticles with diameters as large as 200 nm penetrate the same sputum samples at rates only ∼40-fold reduced compared to in pure water. A limited fraction of AAV exhibit sufficiently fast mobility to penetrate physiologically thick sputum layers, likely because of the lower viscous drag and smaller surface area for adhesion to sputum constituents. Nevertheless, poor penetration of CF sputum is likely a major contributor to the ineffectiveness of viral vector based gene therapy in the lungs of CF patients observed to date

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

ジヘイショウ スペクトラム シャ ノ シンタイ ズシキ ニ カンスル ケンキュウ : テ ノ シンテキ カイテン ニオケル シセイ ノ エイキョウ ニ ツイテ

The purpose of this study was to clarify the influence of the postural condition on the mental rotation task of hands among people with autism spectrum disorder (ASD). Nine people with ASD and seventeen age-matched healthy people participated in this study. Based on a previous developmental study, it was hypothesized that ASD participants would show a larger effect of hand posture than healthy participants. Four line drawings of hands; palm and back view of both left and right hands were presented to the participants on the computer screen at four different rotation angles. (0°,90°,180°, and 270°) The participants were asked to answer whether the presented picture was the left or right hand in the two different hand conditions, the palms-up posture or the backs-up posture. The participants' response time (RT) and the percentage of the correct responses were analyzed by ANOVA. Percentage of the correct response showed that the simple main effect of posture was significant for the ASD but not for healthy group. That is, when participants' hands were in the backs-up posture, the ASD group was more accurate for the back-view stimulus than palm-view stimulus. Concerning RTs, the ASD group tended to respond faster when their posture and the stimulus were matching than when the two were mismatching (p<0.09 for backs-up posture, p< 0.2 for palms-up posture). In contrast, the postural effect on the healthy group was limited to the backs-up posture as reported by the previous research. Thus, in terms of both accuracy and speed, the posture may have a larger effect on people with ASD than healthy people. To show more clear-cut group differences, additional data with more ASD participants are needed

Sample 20 s trajectories.

<p>Representative trajectories of (A) Adenovirus (Adv), (B) AAV5, (C) muco-adhesive PS control nanoparticles and (D) muco-inert control PS-PEG nanoparticles. All trajectories have MSD values within one standard deviation of the ensemble average.</p

Theoretical model of particle penetration across a sputum layer.

<p>(A) <i>S</i>chematic of the model, where particles are deposited in airway lumen on top of the CF sputum layer and must penetrate a 10 µm sputum layer to reach the epithelial cells. The pericilliary layer is much smaller in CF patients due to the collapsed cilia from the accumulated sputum <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0019919#pone.0019919-Boucher2" target="_blank">[60]</a>. (B) Estimated fraction of viral and synthetic particles that are capable of penetrating a 10 µm thick layer of CF sputum over time using Fick's second law. *Statistically significant difference when compared with AAV5, AdV or PS (p<0.05).</p

Transport mode distributions of AdV, AAV5, PS and PS-PEG particles.

<p>Particles were classified into either (i) immobile or hindered and (ii) diffusive <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0019919#pone.0019919-Lai4" target="_blank">[12]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0019919#pone.0019919-Lai5" target="_blank">[15]</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0019919#pone.0019919-Suk2" target="_blank">[17]</a>. *Statistically significant difference when compared with AAV5, AdV or PS within the same transport mode classification (p<0.05).</p