Project description:Transcriptional profiling of U937 scramble vs shHDAC2 before and after SAHA treatment at 5µM concentration for 6 and 24 hours. Different Experimental Conditions: U937 scramble (U937 trasfected with empty vector) vs shHDAC2 (U937 trasfected with shHDAC2 vector), untreated and treated with SAHA at 5 µM concentration for 6h and 24h. Biological replicates: 2 for each sample, independently grown and harvested at 6 and 24 hours. One replicate per array.
Project description:Transcriptional profiling of U937 miR-194-5p (UmiR-194-5p) vs U937 miR-194-5p (UmiR-194-5p) treated with SAHA (Vorinostat; suberoylanilide hydroxamic acid) for 24 h at 5uM concetration
Project description:Design: Persistent latently infected CD4+ T cells represent a major obstacle to HIV eradication. Histone deacetylase inhibitors (HDACis) are a promising activation therapy in a “shock and kill” strategy. However, off-target effects of HDACis on host gene expression are poorly understood in primary cells of the immune system. We hypothesized that HDACi-modulated genes would be best identified with a dose response analysis. Methods: Resting primary CD4+ T cells were treated with increasing concentrations (0.34, 1, 3, or 10 μM) of the HDACi, suberoylanilide hydroxamic acid (SAHA), for 24 hours and then subjected to microarray gene expression analysis. Genes with dose-correlated expression were identified with a likelihood ratio test using Isogene GX and a subset of these genes with a consistent trend of up or downregulation at each dose of SAHA were identified as dose-responsive. Histone modifications were characterized in promoter regions of the top 6 SAHA dose-responsive genes by RT-qPCR analysis of immunopreciptated chromatin (ChIP). Results: A large number of genes were shown to be up (N=657) or down (N=725) regulated by SAHA in a dose-responsive manner (FDR p-value < 0.05 and fold change ≥ |2|). Several of these genes (CTNNAL1, DPEP2, H1F0, IRGM, PHF15, and SELL) are potential in vivo biomarkers of SAHA activity. SAHA dose-responsive gene categories included transcription factors, HIV restriction factors, histone methyltransferases, and host proteins that interact with HIV proteins or the HIV LTR. Pathway analysis suggested net downregulation of T cell activation with increasing SAHA dose. Histone acetylation was not correlated with host expression, but plausible alternative mechanisms for SAHA-modulated expression were identified. Conclusions: Numerous host genes in CD4+ T cells are modulated by SAHA in a dose-responsive manner, including genes that may negatively influence HIV activation from latency. Our study suggests that SAHA influences gene expression through a confluence of several mechanisms, including histone acetylation, histone methylation, and altered expression and activity of transcription factors.
Project description:This study evaluated primary CD4+ T cell gene expression treated with pharmacologically achievable concentration (340 nM) of SAHA for 24 hours in order to evaluate potential side effects of this compound in cells relevant to HIV infection. Analysis of human primary CD4+ T cells taken from 9 healthy donors treated with 340 nM of SAHA for 24 hours. Results identify genes modulated by SAHA treatment in human primary CD4+ T cells.
Project description:To elucidate the molecular mechanism involved in increased tolerance to high-salinity stress by the application of HDAC inhibitor named SAHA
Project description:This study evaluated primary CD4+ T cell gene expression treated with pharmacologically achievable concentration (340 nM) of SAHA for 24 hours in order to evaluate potential side effects of this compound in cells relevant to HIV infection.