Effect of compounds (9, 13, 20 and 21) on the HIV-1 transcription in TZM-bl cells infected with HIV-1NL4-3

We developed a cellular screening system capable of simultaneously evaluating the functional activities of Tat-induced LTR transcription and general cellular expression. Herein, we identified and optimized novel HIV-1 Tat inhibitory compounds that contain an oxadiazole core. To assess the inhibitory effect of our lead compounds on Tat-mediated HIV-1 transcription, we performed RNA-seq analyses of the HIV-1 transcripts in the compound-treated HIV-1-infected cells. The read counts of the transcripts mapped to the HIV-1 genome were substantially reduced in the compound 9- or 13-treated cells; however, the decrease of the read counts was not observed in the HIV-1 infected cells with the ineffective compounds (20 and 21). These data indicate that the viral transcription step is a target for our lead compounds. Overall design: Novel oxadiazole compounds that inhibit HIV-1 Tat-mediated viral transcription were identified. To elucidate the Molecular mechanism underlying the Tat inhibition of compounds, we performed the level of viral transcripts using an RNA-seq analysis gene expression of our lead compounds treated-HIV-1 infected cells. Comparative gene expression profiling analysis of RNA-seq data for DMSO and compounds (9, 13, 20 and 21) treated cells.

本研究构建了可同时评估Tat反式激活蛋白（Trans-Activator of Transcription, Tat）诱导的长末端重复序列（Long Terminal Repeat, LTR）转录功能活性与基础细胞基因表达水平的细胞筛选系统。在此研究中，我们筛选并优化了一类以恶二唑为母核的新型HIV-1 Tat抑制性化合物。为评估先导化合物对Tat介导的HIV-1转录的抑制效果，我们对经化合物处理的HIV-1感染细胞中的HIV-1转录本开展了RNA测序（RNA-seq）分析。经化合物9或13处理的细胞中，比对至HIV-1基因组的转录本读段计数显著降低；而经无效化合物20、21处理的HIV-1感染细胞中则未出现读段计数下降的现象。上述实验数据表明，病毒转录步骤是本研究先导化合物的作用靶点。
实验设计：本研究筛选得到了可抑制HIV-1 Tat介导的病毒转录的新型恶二唑类化合物。为阐明化合物对Tat的抑制作用分子机制，我们通过RNA测序分析检测了经先导化合物处理的HIV-1感染细胞的病毒转录本水平，并开展了基因表达分析；同时对二甲基亚砜（Dimethyl Sulfoxide, DMSO）、化合物9、13、20及21处理的细胞的RNA测序数据开展了比较基因表达谱分析。