Regulatory mechanism of HIV-ltranscription and its therapeutic control

HIV-l转录调控机制及其治疗控制

• 批准号: 16017291
• 负责人: OKAMOTO Takashi
• 金额: $ 9.6万
• 依托单位: NAGOYA CITY UNIVERSTTY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16017291/
• 关键词: HIV; transcriptional regulation; Tat; NF-κB; viral replication; inhibitor; OGG1; maintenance of genome integrity; IKK阻害剤; AP-4; 立体構造; 計算化学; HIV転写活性化; RNA helicase A; 立体構造解析; 分子モデリング; Bioinformatics

In an attempt to identify the cellular target genes for HIV-encocled Tat, we have performed gene expression profile analysis and fund OGG1, the gene encoding an enzyme responsible for the repair of 8-oxo-dG, an damaged Guanine residue due to the action of radieal oxygen species (ROS). Among the as-regulatory elements located within the promoter of OGG1 gene, we have identified AP-4 sites to be responsible for the Tat-mediated OGG1 upregulation. When AP-4 sites were mutated the OGG1 gene expression was upregulated, indicating that AP-4 plays a negative role in OGG1 gene expression, and the effect of Tat was abolished. Using chromatin immunoprecipitation (ChIP) assay we fund that AP-4 binds to these AP-4 sites on the OGG1 promoter and AP-4 was reliesed from the OGG1 promoter upon Tat transduction. We also fund that Tat binds to AP-4 by immunoprecipitation followed by Western blotting. When the amounts of 8-oxo-dG level were measured, we found the dramatic decrease of the 8-oxo-dG upon Tat transduction. These findings indicate a possibility that Tat plays a role in Maintaining the genomic integrity of the HIV-infected cells and HIV proviral DNA.We also examined the effect of a novel inhibitor of IKK that is primarily involved in the signal induced HIV replication from the latently infected cells. The HIV production was greatly augmented by stimulating cells infected with HIV and the pretreatment of cells with ACHP greatly inhibited the viral production under non-cytotoxic concentrations with IC 50 and CC50 values of 0.5 μM and 15 μM, respectively (therapeutic window being approximately 30). These findings suggest that ACHP and its derivatives may have therapeutic effect to prevent AIDS development by preventing HIV replication from the latently infected cells.

为了确定 HIV 包裹的 Tat 的细胞靶基因，我们进行了基因表达谱分析并资助了 OGG1，该基因编码负责修复 8-oxo-dG 的酶，8-oxo-dG 是由于作用而受损的鸟嘌呤残基。在位于 OGG1 基因启动子内的 as 调节元件中，我们发现 AP-4 位点负责 Tat 介导的 OGG1 上调。突变后 OGG1 基因表达上调，表明 AP-4 在 OGG1 基因表达中发挥负面作用，并且使用染色质免疫沉淀 (ChIP) 检测，我们发现 AP-4 与这些 AP-4 位点结合。 OGG1 启动子和 AP-4 依赖于 Tat 转导时的 OGG1 启动子，我们还通过免疫沉淀和蛋白质印迹证实了 Tat 与 AP-4 的结合。测量了 8-oxo-dG 水平，我们发现 Tat 转导后 8-oxo-dG 急剧下降，这些发现表明 Tat 在维持 HIV 感染细胞和 HIV 原病毒的基因组完整性方面发挥着作用。 DNA。我们还检查了一种新型 IKK 抑制剂的作用，该抑制剂主要参与信号诱导潜伏感染细胞的 HIV 复制。通过刺激感染 HIV 的细胞和用 ACHP 预处理细胞，大大增强了 HIV 的产生。在非细胞毒性浓度下极大地抑制病毒产生，IC 50 和 CC50 值分别为 0.5 μM 和 15 μM（治疗窗口约为 30）。这些发现表明 ACHP 及其衍生物可能具有预防艾滋病发展的治疗作用。通过阻止潜伏感染细胞中的艾滋病毒复制。

项目成果

Growth inhibition o multiple myeloma cells by a novel IkB kinase inhibitor.

新型 IkB 激酶抑制剂抑制多发性骨髓瘤细胞的生长。

• 发表时间: 2005
• 期刊: Clin.Can.Res. 11
• 作者: Sanda;T.
• 通讯作者: T.

NF-kB signaling an carcinogenesis.

NF-kB 发出致癌信号。

• 发表时间: 2006
• 期刊: Curr.Pharm.Design. (In press)
• 作者: Okamoto;T.
• 通讯作者: T.

Positive correlation between sialyl Lewis X expression and pathological findings in renal cell carcinoma.

肾细胞癌中唾液酸Lewis X表达与病理结果呈正相关。

• 发表时间: 2005
• 期刊: Kidney Int. 67
• 作者: Tozawa;K.
• 通讯作者: K.

厳選辞書シリーズ「転写」エロンゲータ SII エロンギン P-TEFb CSB ELL DSIF NELF

严选词典系列《转录》 Elongator SII Elongin P-TEFb CSB ELL DSIF NELF

• 发表时间: 2006
• 作者: Okamoto;T.;岡本 尚;岡本 尚
• 通讯作者: 岡本 尚

Positive correlation between sialyl Lewis X expression ani pathological findings in renal cell carcinoma.

肾细胞癌中唾液酸Lewis X表达与病理结果呈正相关。

• 发表时间: 2005
• 期刊: Kidney Int. 67
• 作者: Tozawa;K.
• 通讯作者: K.