Journal: Translational Psychiatry

Article Title: Decreased AGO2 and DCR1 in PBMCs from War Veterans with PTSD leads to diminished miRNA resulting in elevated inflammation

doi: 10.1038/tp.2017.185

Figure Lengend Snippet: miRNA expression profile in PBMCs from post-traumatic stress disorder (PTSD) patients. ( a ) Microarray analysis of miRNAs in PBMCs from normal human controls and PTSD patients. The heat map shows the expression intensities of all the miRNAs in each of the control and PTSD samples after microarray. ( b ) The volcano plot shows all the miRNAs with at least ±1.5 linear fold-difference. The green dots represents miRNAs significantly dysregulated in PTSD patients. (On y axis, 1.301 units is ~0.05). A total of 190 miRNAs were found to be significantly ( P- value<0.05) up- (7 miRNAs) or downregulated (183 miRNAs) by a linear fold-change value of ±1.5 or more ( c ). ( d ) The expression levels of the precursors of miRNAs as obtained from RNA-Seq analysis and presented here in the figure as fold-change values. The names represent the corresponding precursors of the miRNAs. These 35 miRNAs were further analyzed in the in vitro studies. PBMC, peripheral blood mononuclear cell.

Article Snippet: Microarray for the miRNAs was performed at Johns Hopkins Memorial Institute (Deep Sequencing and Microarray Core Facility, Baltimore, MD, USA).

Techniques: Expressing, Microarray, Control, RNA Sequencing, In Vitro

Journal: Translational Psychiatry

Article Title: Decreased AGO2 and DCR1 in PBMCs from War Veterans with PTSD leads to diminished miRNA resulting in elevated inflammation

doi: 10.1038/tp.2017.185

Figure Lengend Snippet: Argonaute 2 ( AGO2 ) and Dicer1 ( DCR1 ) transcript is lower in PBMCs of post-traumatic stress disorder (PTSD) patients and the abundance of mature miRNAs is reduced upon decreased expression of AGO2 and DCR1. ( a ) RNA-Seq analysis expression values of only the genes significantly dysregulated with log 2 fold-change of at least 1 or more (readers are requested to refer Bam et al. for the complete list of the genes), and different variants of AGO2 , DCR1 and STAT3 . The green dots indicate the position (expression values) of the different variants of AGO2 , DCR1 and STAT3 . The red dot is the position of DCR1 variant 5. ( b ) The FPKM values of the variants of AGO2 and DCR1 in control and PTSD samples after RNA-sequencing (RNA-Seq) analysis. The values above the bars indicate log 2 fold-change when compared between PTSD and controls. (FPKM: Fragments per kilobase of transcript per million mapped reads). ( c , d ) Quantitative real time PCR validation result of AGO2 and DCR1 transcripts in 22 controls and 18 PTSD PBMC RNA samples. The difference in expression level is provided as relative expression (RE) value by taking the controls as 1. In this assay, 18S rRNA was used as an internal control. ( e ) Linear fold-change values of 35 miRNAs, after microarray analysis, which were selected for further analysis by in vitro experiments. miRNA-451 was also included in the figure as a positive control because it was reported previously to be processed specifically through the AGO2-dependent pathway. ( f ) Relative expression levels of 34 miRNAs, listed in Figure 3a, after knockdown of AGO2 for 72 h by employing siRNA in THP-1 cells. The expression level is expressed relative to control which was taken as 1. PBMC, peripheral blood monnuclear cell.

Article Snippet: Microarray for the miRNAs was performed at Johns Hopkins Memorial Institute (Deep Sequencing and Microarray Core Facility, Baltimore, MD, USA).

Techniques: Expressing, RNA Sequencing, Variant Assay, Control, Real-time Polymerase Chain Reaction, Biomarker Discovery, Microarray, In Vitro, Positive Control, Knockdown

Journal: Translational Psychiatry

Article Title: Decreased AGO2 and DCR1 in PBMCs from War Veterans with PTSD leads to diminished miRNA resulting in elevated inflammation

doi: 10.1038/tp.2017.185

Figure Lengend Snippet: miRNAs downregulated in post-traumatic stress disorder (PTSD) target several pro-inflammatory genes. ( a ) Ingenuity Pathway Analysis (IPA) was performed with the dysregulated miRNAs in PTSD. The genes were included in the network and the miRNAs were connected by the IPA algorithm. The network shows direct interaction (predicted and proven) between miRNAs and target genes as indicated by solid arrows. ( b ) Principal component analysis (PCA) plot showing the relatedness of the samples. ( c ) IPA-generated network showing the interaction between miRNAs and targets after analyzing with the miRNAs obtained in the replicate samples. The red and blue colored target genes in the network correspond to up- and downregulated, respectively, validated for its expression level and reported either in the current manuscript or in our previous reports with studies on the same samples. For the miRNAs, the red color indicates upregulated and green indicates downregulated as per microarray analysis. ( d ) To validate the transcript levels of miRNA-targeted genes, qRT-PCR was performed to detect transcript of eight genes as shown in the graph (genes were selected based on the miRNA–gene interaction networks). The P- values of the genes under the curly brackets were <0.05.

Article Snippet: Microarray for the miRNAs was performed at Johns Hopkins Memorial Institute (Deep Sequencing and Microarray Core Facility, Baltimore, MD, USA).

Techniques: Generated, Expressing, Microarray, Quantitative RT-PCR