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      Transcriptome organization of white blood cells through gene co-expression network analysis in a large RNA-seq dataset

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          Abstract

          Gene co-expression network analysis enables identification of biologically meaningful clusters of co-regulated genes (modules) in an unsupervised manner. We present here the largest study conducted thus far of co-expression networks in white blood cells (WBC) based on RNA-seq data from 624 individuals. We identify 41 modules, 13 of them related to specific immune-related functions and cell types (e.g. neutrophils, B and T cells, NK cells, and plasmacytoid dendritic cells); we highlight biologically relevant lncRNAs for each annotated module of co-expressed genes. We further characterize with unprecedented resolution the modules in T cell sub-types, through the availability of 95 immune phenotypes obtained by flow cytometry in the same individuals. This study provides novel insights into the transcriptional architecture of human leukocytes, showing how network analysis can advance our understanding of coding and non-coding gene interactions in immune system cells.

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          Most cited references71

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          WGCNA: an R package for weighted correlation network analysis

          Background Correlation networks are increasingly being used in bioinformatics applications. For example, weighted gene co-expression network analysis is a systems biology method for describing the correlation patterns among genes across microarray samples. Weighted correlation network analysis (WGCNA) can be used for finding clusters (modules) of highly correlated genes, for summarizing such clusters using the module eigengene or an intramodular hub gene, for relating modules to one another and to external sample traits (using eigengene network methodology), and for calculating module membership measures. Correlation networks facilitate network based gene screening methods that can be used to identify candidate biomarkers or therapeutic targets. These methods have been successfully applied in various biological contexts, e.g. cancer, mouse genetics, yeast genetics, and analysis of brain imaging data. While parts of the correlation network methodology have been described in separate publications, there is a need to provide a user-friendly, comprehensive, and consistent software implementation and an accompanying tutorial. Results The WGCNA R software package is a comprehensive collection of R functions for performing various aspects of weighted correlation network analysis. The package includes functions for network construction, module detection, gene selection, calculations of topological properties, data simulation, visualization, and interfacing with external software. Along with the R package we also present R software tutorials. While the methods development was motivated by gene expression data, the underlying data mining approach can be applied to a variety of different settings. Conclusion The WGCNA package provides R functions for weighted correlation network analysis, e.g. co-expression network analysis of gene expression data. The R package along with its source code and additional material are freely available at .
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            Differential expression analysis for sequence count data

            High-throughput sequencing assays such as RNA-Seq, ChIP-Seq or barcode counting provide quantitative readouts in the form of count data. To infer differential signal in such data correctly and with good statistical power, estimation of data variability throughout the dynamic range and a suitable error model are required. We propose a method based on the negative binomial distribution, with variance and mean linked by local regression and present an implementation, DESeq, as an R/Bioconductor package.
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              g:Profiler: a web server for functional enrichment analysis and conversions of gene lists (2019 update)

              Abstract Biological data analysis often deals with lists of genes arising from various studies. The g:Profiler toolset is widely used for finding biological categories enriched in gene lists, conversions between gene identifiers and mappings to their orthologs. The mission of g:Profiler is to provide a reliable service based on up-to-date high quality data in a convenient manner across many evidence types, identifier spaces and organisms. g:Profiler relies on Ensembl as a primary data source and follows their quarterly release cycle while updating the other data sources simultaneously. The current update provides a better user experience due to a modern responsive web interface, standardised API and libraries. The results are delivered through an interactive and configurable web design. Results can be downloaded as publication ready visualisations or delimited text files. In the current update we have extended the support to 467 species and strains, including vertebrates, plants, fungi, insects and parasites. By supporting user uploaded custom GMT files, g:Profiler is now capable of analysing data from any organism. All past releases are maintained for reproducibility and transparency. The 2019 update introduces an extensive technical rewrite making the services faster and more flexible. g:Profiler is freely available at https://biit.cs.ut.ee/gprofiler.
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                Author and article information

                Contributors
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                Journal
                Front Immunol
                Front Immunol
                Front. Immunol.
                Frontiers in Immunology
                Frontiers Media S.A.
                1664-3224
                02 April 2024
                2024
                : 15
                : 1350111
                Affiliations
                [1] 1 Istituto di Ricerca Genetica e Biomedica (IRGB), Consiglio Nazionale delle Ricerche (CNR) , Cagliari, Italy
                [2] 2 CRS4-Next Generation Sequencing (NGS) Core, Parco POLARIS , Cagliari, Italy
                [3] 3 Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health (NIH) , Baltimore, MA, United States
                [4] 4 Dipartimento di Medicina Traslazionale e di Precisione, Università Sapienza , Roma, Italy
                [5] 5 Dipartimento di Scienze Biomediche, Università degli Studi di Sassari , Sassari, Italy
                Author notes

                Edited by: Issam El Naqa, University of Michigan, United States

                Reviewed by: Nitin Khandelwal, University of Texas Southwestern Medical Center, United States

                Pei Shang, Mayo Clinic, United States

                *Correspondence: Paola Forabosco, paola.forabosco@ 123456cnr.it
                Article
                10.3389/fimmu.2024.1350111
                11018966
                38629067
                15589795-0af8-450b-9e80-b1b801021f9b
                Copyright © 2024 Forabosco, Pala, Crobu, Diana, Marongiu, Cusano, Angius, Steri, Orrù, Schlessinger, Fiorillo, Devoto and Cucca

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 06 December 2023
                : 13 March 2024
                Page count
                Figures: 3, Tables: 6, Equations: 0, References: 71, Pages: 18, Words: 9535
                Funding
                The author(s) declare financial support was received for the research, authorship, and/or publication of this article. The SardiNIA project is supported in part by the intramural program of the National Institute on Aging through contract HHSN271201100005C to the Consiglio Nazionale delle Ricerche (CNR) of Italy.
                Categories
                Immunology
                Original Research
                Custom metadata
                Systems Immunology

                Immunology
                immune system,network analysis,wgcna,lncrna,rna-seq,white blood cells
                Immunology
                immune system, network analysis, wgcna, lncrna, rna-seq, white blood cells

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