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

Application of Finite Element Method to Analyze the Influences of Process Parameters on the Cut Surface in Fine Blanking Processes by Using Clearance-Dependent Critical Fracture Criteria

The correct choice of process parameters is important in predicting the cut surface and obtaining a fully-fine sheared surface in the fine blanking process. The researchers used the value of the critical fracture criterion obtained by long duration experiments to predict the conditions of cut surfaces in the fine blanking process. In this study, the clearance-dependent critical ductile fracture criteria obtained by the Cockcroft-Latham and Oyane criteria were used to reduce the time and cost of experiments to obtain the value of the critical fracture criterion. The Finite Element Method (FEM) was applied to fine blanking processes to study the influences of process parameters such as the initial compression, the punch and die corner radii and the shape and size of the V-ring indenter on the length of the sheared surface. The effects of stress triaxiality and punch diameters on the cut surface produced by the fine blanking process are also discussed. The verified process parameters and tool geometry for obtaining a fully-fine sheared SPCC surface are described. The results showed that the accurate and stable prediction of ductile fracture initiation can be achieved using the Oyane criterion

Determination of the Values of Critical Ductile Fracture Criteria to Predict Fracture Initiation in Punching Processes

Punching processes are widely used for producing automobile parts, mechanical components, and other parts. To produce highly accurate parts, it is important to estimate the ratio of the sheared surface to the cut surface. Many researchers have applied the finite-element method (FEM) to analyze the ratio of the sheared surface to the fracture surface on cut surfaces by using ductile fracture criteria. However, it is difficult to determine the fracture criteria on the cut surface by tensile tests or bending tests because the punching process involves many complicated steps. In this study, FEM was applied to the punching process to determine the values of critical fracture criteria (C) by using the ductile fracture criteria proposed by Cockcroft and Latham, Oyane, and Ayada. The ductile fracture criteria were compared with the boundary between the shear surface and the fracture surfaces using experiments performed with a simple punching system. The values of the ductile fracture criteria for the fracture initiation of the formed cut surface were predicted under various clearances between the punch and the die with various punch diameters. The influence of stress triaxiality and the effect of punch diameter on the sheared surface length are also discussed

A Novel Method to Reduce ELISA Serial Dilution Assay Workload Applied to SARS-CoV-2 and Seasonal HCoVs

Assays using ELISA measurements on serially diluted serum samples have been heavily used to measure serum reactivity to SARS-CoV-2 antigens and are widely used in virology and elsewhere in biology. We test a method using Bayesian hierarchical modelling to reduce the workload of these assays and measure reactivity of SARS-CoV-2 and HCoV antigens to human serum samples collected before and during the COVID-19 pandemic. Inflection titers for SARS-CoV-2 full-length spike protein (S1S2), spike protein receptor-binding domain (RBD), and nucleoprotein (N) inferred from 3 spread-out dilutions correlated with those inferred from 8 consecutive dilutions with an R2 value of 0.97 or higher. We confirm existing findings showing a small proportion of pre-pandemic human serum samples contain cross-reactive antibodies to SARS-CoV-2 S1S2 and N, and that SARS-CoV-2 infection increases serum reactivity to the beta-HCoVs OC43 and HKU1 S1S2. In serial dilution assays, large savings in resources and/or increases in throughput can be achieved by reducing the number of dilutions measured and using Bayesian hierarchical modelling to infer inflection or endpoint titers. We have released software for conducting these types of analysis