Differential expression of LncRNAs and mRNAs and their modulated network construction during the process of HIV transcription
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ABSTRACT: Objective: Seek the differentially expressed mRNAs and long non-coding RNAs (LncRNAs) to construct the modulated network and seek for the hub LncRNAs during the process of human immunodeficiency virus (HIV) transcription. Method: Blood samples from HIV-infections were collected. They were divided into transcription active group and relatively inactive group based on the quantitative detection results of total HIV DNA and cell-association RNA (CA-RNA) from peripheral blood mononuclear cells (PBMCs). Individuals within each group were matched by age and genders. Microarray chips detection technology were utilized to detect and find the differentially expressed LncRNAs and mRNAs by statistical method of paired T test, then underwent some bioinformatics analysis. Result: 1240 differentially expressed genes (include 689 LncRNAs and 551 mRNAs) were found. Gene ontology analysis showed the differentially expressed mRNAs mainly enriched in positive or negative gene expression, immune cell formation or epigenetic regulation in the biological process; nucleosome, nucleolus, cytoplasm, nuclear chromatin in the cell component; RNA polymerase II regulatory region sequence-specific DNA binding, DNA, histone, and nucleic acid binding, transcription factor activity in molecular function. Pathway analysis showed the differentially expressed mRNAs enriched in the biological pathways such as systemic lupus erythematosus, alcoholism, cytokine-cytokine receptor interaction, RNA transport, mRNA surveillance pathway, Pathways in cancer, etc. Cis and trans analysis to the differentially expressed LncRNAs searching its targeted genes and then made intersection for the differentially expressed mRNAs, then constructed the modulated network between LncRNAs and mRNAs. Based on this modulated network, four hub transcripts were excavated and validated by quantitative real-time PCR to confirm the veracity of microarray analysis findings. Two hub transcripts of lnc-CCNYL1-1:2 and ZNF793 were found directly interacted in the modulated network and their level of expression transformed parallelly. Conclusion: Quantitative detection results of cellular levels of total HIV DNA and RNA can distinguish whether HIV is in the state of active transcription. During process of HIV transcription, the differentially expressed LncRNAs involve the biological process of gene expression regulation, the hub genes excavated form the modulated network may become the potential candidate targets for HIV antiviral treatment.
Project description:Studying the effects of HIV infection on the host transcriptome has typically focused on protein-coding genes. However, recent advances in the field of RNA sequencing revealed that long non-coding RNAs (lncRNAs) add an extensive additional layer to the cell’s molecular network and are highly involved in the regulation of molecular processes, including gene expression. Here, we performed transcriptome profiling throughout a primary HIV infection in vitro to investigate lncRNA expression at the different HIV replication cycle processes (reverse transcription, integration and particle production). Subsequently, guilt-by-association, transcription factor and co-expression analysis were performed to infer biological roles for the lncRNAs identified in the HIV-host interplay. Many lncRNAs were shown to play a role in mechanisms relying on proteasomal and ubiquitinylation pathways, apoptosis, DNA damage responses and cell cycle regulation. Through transcription factor binding analysis, we found that lncRNAs display a distinct transcriptional regulation profile as compared to protein coding mRNAs, suggesting that mRNAs and lncRNAs are independently modulated from each other. In addition, we identified five differentially expressed lncRNA-mRNA pairs with mRNA involvement in HIV pathogenesis (HIV linked co-expression) with possible cis regulatory lncRNAs that control the nearby mRNA expression and function. Altogether, the present study demonstrates that lncRNAs add a new dimension to the HIV-host interplay and should be further investigated as they may represent targets for controlling HIV replication.
Project description:Tripterygium glycosides (TG) was reported to have effect of ameliorating Alzheimer's disease (AD). However, the mechanism is not clear. We aimed to investigate the lncRNAs and circRNAs expression profiles of AD treated with TG by using microarray. LncRNAs, mRNA and circRNA in 3 AD mice and 3 AD+TG mice hippocampal were detected by microarray. The most differentially expressed lncRNAs, mRNA and circRNA in AD+TG group were screened. The differentially expressed lncRNAs and circRNAs were analyzed for GO enrichment and KEGG pathway. Co-expression analysis of lncRNAs and mRNA was performed by calculating correlation coefficients. Protein-protein interaction (PPI) network analysis was performed on mRNAs using STRING. LncRNA-target-TFs network were analyzed by Network software. CircRNA-mirNA network were conducted by Cytoscape software. A total of differentially expressed 661 lncRNAs, 64 circRNAs and 503 mRNAs were detected in AD mice treated with TG. The Pou4f1, Egr2, Mag, and Nr4a1 were the hub genes in the PPI network. The KEGG results showed the mRNAs that co-expressed with lncRNAs were enriched in TNF, PI3K-Akt, and Wnt signaling pathway. lncRNA-target-TFs network analysis indicated the TFs including Cebpa, Zic2, and Rxra were the most likely to regulate the detected lncRNAs. The circRNA-miRNA interaction network indicated 275 miRNAs may bind to the 64 circRNAs. In conclusion, the findings supplied a novel perspective for AD pathogenesis. And the detected lncRNAs, mRNAs, and circRNAs might be novel therapeutics targets for AD.
Project description:We identified the differentially expressed mRNAs and lncRNAs/circRNAs of the case group and the control group through high-throughput sequencing technology, performed a functional enrichment analysis of the differentially expressed genes, and used protein-protein interaction (PPI) analysis to identify the hub genes. A novel ceRNA network was successfully established, and the network components may serve as promising diagnostic biomarkers or therapeutic targets for thyroid-associated ophthalmopathy in the future.
Project description:This study aimed to identify the crucial molecules and explore the function of noncoding RNAs and related pathways in IDD. We randomly selected 3 samples each from an IDD and a spinal cord injury group (control) for RNA-sequencing. We identified 463 differentially-expressed long noncoding RNAs (lncRNAs), 47 differentially-expressed microRNAs (miRNAs), and 1,334 differentially-expressed mRNAs in IDD. Three hundred fifty-eight lncRNAs as cis-regulators could potentially target 865 genes. Protein–protein interaction (PPI) network analysis confirmed that IL-6, VEGFA, IGF1, MMP9, CXCL8, FGF2, IL1B, CCND1, ITGAM, PTPRC, FOS and PTGS2 were hub genes. We built a competing endogenous RNA (ceRNA) network and identified lncRNA XIST–hsa-miR-4775–PLA2G7 and lncRNA XIST–hsa-miR-424-5p–AMOT/TGFBR3 ceRNA axes.
Project description:Genome-wide analysis of skin color-related lncRNA and mRNA expression in Koi carp, Cyprinus carpio L. LncRNAs information linked to fish skin color regulation is over-limited. In this study, Illumina sequencing and bioinformatics were primarily conducted on black, white and red skin colors of Koi carp. A total of 590,415,050 clean reads, 446,614 putative transcripts, 4,252 known and 72,907 novel lncRNAs were simultaneously obtained, respectively. Out of these genes, 92 significant differentially expressed lncRNAs and 722 mRNAs were excavated. Ccr_lnc5622441, Ccr_lnc765201 were found up-regulated in black and red skins; Ccr_lnc14074601 were up-regulated in white skin; and premelanosome proteins a (Pmela), tyrosinase (Tyr) were up-regulated in black skin, etc. Quantitative real-time PCR (qRT-PCR) further validated 12 differentially expressed genes were consistent with RNA-seq. Moreover, 70 lncRNAs on 107 target mRNAs in cis and 79 lncRNAs on 41,625 target mRNAs in trans were investigated, the networks revealed one lncRNAs can connected with numerous mRNAs, vice versa. These findings broadened the lncRNAs landscape of skin colors and provided new insights into the mechanisms underlying lncRNAs mediated pigmentation and differentiation in Koi carp.
Project description:Chlamydia trachomatis (C. trachomatis) is a major etiological agent of sexually transmitted infection. Some stressing conditions can result in persistent chlamydial infection, which is thought to associate with severe complications such as ectopic pregnancy and tubal factor infertility. Long noncoding RNAs (lncRNAs) have been identified as key modulators in many biological processes. However, the role of lncRNAs in persistent chlamydial infection is still unclear. In this study, we used lncRNA and mRNA microarray to identify the global lncRNAs and mRNAs expression in penicillin-induced persistent chlamydial infection in HeLa cells as well as the control group (HeLa cells without C. trachomatis infection). Among 1005 differentially expressed lncRNAs, 585 lncRNAs were upregulated and 420 downregulated in persistent chlamydial infection, while 410 mRNAs were identified to express differentially, of which 113 mRNAs were upregulated and 297 downregulated. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis with differentially expressed genes were performed. We then constructed the lncRNA-miRNA-mRNA competing endogenous RNAs (ceRNAs) network. Four mRNAs were validated to be changed by quantitative real-time PCR which were correlated with the microarray result. Integration of protein-protein interaction (PPI) network was constructed and hub genes were identified. These findings provide a new perspective on the molecular mechanism of penicillin-induced persistent chlamydial infection.
Project description:Studies have shown that long noncoding RNAs (lncRNAs) can be widely involved in various physiological and pathological processes. In recent years, there have been many studies on GLP-1 receptor agonists (GLP-1RA) regulating islet β cells function and mass of type 2 diabetes (T2DM) patients. However, the function of lncRNAs in this process have not been fully elucidated. In this study, lncRNA microarray was used to identify the differently expressed (DE) lncRNAs and mRNAs in β cells exposed to Geniposide, which is a GLP-1RA. 308 lncRNAs and 128 mRNAs were detected with a set filter fold change≥1.5 and P-value<0.05. Gene Ontology and KEGG pathway analysis were performed to assess the underlying functions of DE mRNAs. Co-expression network of DE lncRNAs and mRNAs was constructed based on Pearson coefficient of expression level. And hub mRNAs were selected through String database and Cytoscape plugin Cytohubba. Additionally, a ceRNA network was constructed among the co-expressed lncRNAs and hub mRNAs. This study reveals key mRNAs involved in the regulation of GLP-1RA on β cells function and mass. More importantly, screening out lncRNAs that play an importance regulatory role in this process. This original research could provide valuable information for the further investigation between lncRNAs and GLP-AR in the protection of β cells.
Project description:Transcriptome of human retinal endothelial cells (hRECs) treated with low glucose (LG), high glucose (HG) or high glucose with 4 uM TTR (HG+TTR) were conducted. Differentially expressed lncRNAs, mRNAs and TTR related lncRNAs and mRNA were acquired for further analysis. Functional annotation and enrichment including KEGG pathway, GO and GSEA were applied to analyze TTR regulated pathway and process. WGCNA analysis was implemented to obtain hub modules and genes. LncRNA-mRNA regulatory network were constructed based on cis, trans and ceRNA acting mode.