Simultaneous latent HIV reactivation and cytokine gene suppression

描述（由申请人提供）：潜伏HIV-1的持续存在仍然是根除感染的治疗工作的主要挑战。之前清除病毒库的尝试 包括T细胞有丝分裂原或组蛋白脱乙酰酶抑制剂（HDACIs），在减少病毒库方面的成功有限，强调了对促进病毒再活化的其他方法的需要。这一提议的动机是我们最近的发现[1]，即最初被鉴定为有效的溴结构域-4（Brd 4）拮抗剂和癌症抑制剂的小分子JQ 1，有效地上调细胞系和原代CD 4 + T细胞中潜伏的和诱导的HIV前病毒表达。此外，JQ 1抑制急性暴露于内毒素的小鼠的全身炎症细胞因子产生，并抑制原代CD 4 + T细胞中的T细胞活化基因。因此，JQ 1代表了具有所需特征和翻译效用的新型化合物。我们的总体假设是JQ 1通过调节Brd 4活性促进病毒再活化并抑制NF-κ B驱动的基因表达。作为干扰HIV启动子处Brd 4活性的结果，JQ 1可增加病毒达特蛋白对宿主正转录延伸因子（P-TEFb）的接近。在细胞因子基因启动子处，我们假设JQ 1干扰乙酰化p65/RelA与Brd 4的相互作用，导致NF- B驱动的细胞因子基因表达下降，但不影响NFAT驱动的基因表达。为了解决我们的假设，我们提出了一个翻译程序，以表征该化合物对HIV前病毒再激活和对潜伏T细胞系中细胞因子基因表达的影响，该细胞系是原代T细胞的体外潜伏模型， 来自接受有效ART的感染个体的静息CD 4 + T细胞。

DESCRIPTION (provided by applicant): The persistence of latent HIV-1 remains a major challenge in therapeutic efforts to eradicate infection. Prior attempts at purging viral reservoirs have included T cell mitogens or histone deacetylase inhibitors (HDACIs) with limited success in reducing the viral reservoir, underscoring the need for additional approaches that promote viral reactivation. This proposal is motivated by our recent discovery [1] that a small molecule, JQ1, initially identified as a potent bromodomain-4 (Brd4) antagonist and cancer suppressor, potently upregulates latent and induced HIV proviral expression in cell lines and primary CD4+ T cells. In addition, JQ1 suppresses systemic inflammatory cytokine production in mice acutely exposed to endotoxin and suppresses T cell activation genes in primary CD4+ T cells. JQ1 therefore represents a novel compound with desirable features and translational utility. Our overall hypothesis is that JQ1 promotes viral reactivation and suppresses NF-kB driven gene expression by modulating Brd4 activity. As a result of interfering with Brd4 activity at the HIV promoter, JQ1 may increase access of the viral Tat protein to the host positive transcriptional elongation factor (P-TEFb). At cytokine gene promoters, we hypothesize that JQ1 interferes with acetylated p65/RelA interaction with Brd4 resulting in declines in NF- B driven cytokine gene expression, but not NFAT driven gene expression. To address our hypothesis, we propose a translational program to characterize this compound's effect on HIV proviral reactivation and on cytokine gene expression in latent T cell lines, an in vitro latency model for primary T cells and in resting CD4+ T cells from infected individuals receiving effective ART.