27 68 miRNAs were dysregulated somehow greater than 2 fold. Hierarchical Inhibitors,Modulators,Libraries clustering was carried out to demonstrate patterns of miRNA ex pression profiling between two Inhibitors,Modulators,Libraries groups, which was consistent with our mRNA hierarchical clustering. TargetScan was used to predict the gene tar gets of DE miRNAs, because it is the most advanced, respected, widely used and relatively conservative data base in comparison to other databases. In addition, we have carried out a low and high strin Inhibitors,Modulators,Libraries gency G seed search for miR 137, miR 153 and miR 218 targets based on a minimal free energy ? 10 and ? 14, respectively. Furthermore, these predictions were made in the context of changes in gene expression observed in the same RNA.
Functional annotation of the mRNA DE genes and miRNA target genes was carried out in Database for An notation, Visualization and Integrated Discovery together with extensive literature search. These mRNA DE genes were significantly associated with biological pro cesses, such as cell death cell cycle, neuronal processes, metabolism, transcriptional regulation, protein modifica tion, Inhibitors,Modulators,Libraries signal transduction, and response to virus stress, as shown in Figure 1A. In addition, according to cellular components distribution, 29% of genes fell into neuronal related components, such as axon, neuron projection, and dendrite. Furthermore, Gene Set Enrich ment Analysis was also used to examine signifi cantly enriched GO gene sets comparing the normalized data of the entire 48,701 gene transcripts from HAD and HIV non dementia brains to 1454 GO gene sets in GSEA Molecular Signatures Database.
Eight gene sets were found statistical sig nificantly enriched in HIV non dementia group while no gene sets were significantly Inhibitors,Modulators,Libraries enriched in the HAD group. Of the eight enriched gene sets in the non dementia group, four were closely related to neurological and or HIV disease, den drite, ATPase activity coupled to transmembrane movement of ions, ATPase activity coupled to transmem brane movement of ions phosphorylative mechanism, and cytoskeleton dependent intracellular transport. Functional annotation results of miRNA target genes were consistent with that of mRNA results, but note worthy is that the miRNA target genes showed relatively more comprehensive biological processes and cellular components, which could be attributed to the ability of single miRNA to have numerous mRNA targets, therefore further validation might be needed to precise the specifi city.
Cellular components of miRNA target genes are mainly distributed in cellular and neuronal related structures, but they display nothing scattering into ion channel, actin cytoskeleton, tight junction, tran scription factor complex and chromatin, which concur with our mRNA results. For instance, we found significant dysregulation of genes in microtubule assembly, microtubule nucleation, cyto skeleton movement and microtubule stabilization.