Identification of Small Molecule Inhibitors of Jumonji AT-rich Interactive Domain 1B (JARID1B) Histone Demethylase by a Sensitive High Throughput Screen
Identification of Small Molecule Inhibitors of Jumonji AT-rich Interactive Domain 1B (JARID1B) Histone Demethylase by a Sensitive High Throughput Screen
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DOI：
10.1074/jbc.m112.419861
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发表时间：
2013-03-29
影响因子：
4.8
通讯作者：
Yan, Qin
中科院分区：
文献类型：
--
作者：
Sayegh, Joyce;Cao, Jian;Yan, Qin
JARID 1B（也称为KDM 5 B或PLU 1）是组蛋白赖氨酸脱甲基酶JARID 1家族的成员，负责组蛋白H3（H3 K4 me 3）中三甲基化赖氨酸27的脱甲基化，这是一种活跃转录基因的标记。JARID 1B在多种癌症中过表达，包括乳腺癌、前列腺癌和肺癌。此外，JARID 1B是同基因或异种移植小鼠模型中乳腺肿瘤形成所必需的。表达JARID 1B的黑色素瘤细胞与增加的自我更新特征相关。因此，JARID 1B代表了癌症治疗的有吸引力的靶标。在这里，我们的特点JARID 1B使用均匀发光为基础的脱甲基酶测定。然后，我们对超过15，000个小分子进行了高通量筛选，以鉴定JARID 1B的抑制剂。从这个屏幕上，我们确定了几个已知的JmjC组蛋白去甲基化酶抑制剂，包括2，4-吡啶二羧酸和儿茶酚。更重要的是，我们鉴定了几种新的抑制剂，包括2-4（4-甲基苯基）-1，2-苯并异噻唑-3（2 H）-酮（PBIT），其在体外抑制JARID 1B，IC 50约为3 μ M。与此一致，PBIT处理抑制细胞中JARID 1B对H3 K4 me 3的去除。此外，该化合物抑制表达较高水平JARID 1B的细胞的增殖。这些结果表明，这种新型小分子抑制剂是一种先导化合物，可以进一步优化癌症治疗。
JARID1B (also known as KDM5B or PLU1) is a member of the JARID1 family of histone lysine demethylases responsible for the demethylation of trimethylated lysine 27 in histone H3 (H3K4me3), a mark for actively transcribed genes. JARID1B is overexpressed in several cancers, including breast cancer, prostate cancer, and lung cancer. In addition, JARID1B is required for mammary tumor formation in syngeneic or xenograft mouse models. JARID1B-expressing melanoma cells are associated with increased self-renewal character. Therefore, JARID1B represents an attractive target for cancer therapy. Here we characterized JARID1B using a homogeneous luminescence-based demethylase assay. We then conducted a high throughput screen of over 15,000 small molecules to identify inhibitors of JARID1B. From this screen, we identified several known JmjC histone demethylase inhibitors, including 2,4-pyridinedicarboxylic acid and catechols. More importantly, we identified several novel inhibitors, including 2-4(4-methylphenyl)-1,2-benzisothiazol-3(2H)-one (PBIT), which inhibits JARID1B with an IC50 of about 3 mu M in vitro. Consistent with this, PBIT treatment inhibited removal of H3K4me3 by JARID1B in cells. Furthermore, this compound inhibited proliferation of cells expressing higher levels of JARID1B. These results suggest that this novel small molecule inhibitor is a lead compound that can be further optimized for cancer therapy.