Liu Yan, Platchek Michael, Kement Burcu, Bee Weilin T, Truong Maggie, Zeng Xin, Hung Sunny, Lin Hong, Morrow Dwight, Kallal Lorena A, Xie Qing, Agarwal Pankaj, Pope Andrew J, Wu Zining
Molecular Discovery Research, GlaxoSmithKline, Collegeville, PA, USA.
Mol Biosyst. 2014 Feb;10(2):251-7. doi: 10.1039/c3mb70413k.
Epigenetic regulation by histone methylation is crucial for proper programming of the genome during development. Homeostasis of histone methylation is balanced by the activities of histone methyltransferases and demethylases. Although these methyltransferases and demethylases represent logical targets for potential drug discovery, the activities of methyltransferases and demethylases regulated in response to a complex biological stimulus are also important and not yet clear. To manipulate and study histone methylation in biological systems, we screened a Biologically Diverse Compound Set (BDCS) utilizing a phenotypic assay system that directly measures the Histone 3 K27 tri-methylation (H3K27me3) level in cells. The BDCS is a unique set of target-annotated chemical probes, containing a total of 5853 compounds targeting 736 unique proteins with multiple maximally selective compounds for each target. A number of targets, with multiple hits against each target, were identified in the screen. This gave us confidence that these targets and pathways may be relevant, and included the identification of non-methyltransferase/demethylase targets as potential upstream regulators of H3K27me3. Our study suggests that a systematically designed chemical probe library can serve as a powerful drug discovery tool when combined with phenotypic screening. Follow-up studies using these findings may reveal novel therapeutically useful pathways and targets of H3K27me3 regulation.
组蛋白甲基化介导的表观遗传调控对于发育过程中基因组的正确编程至关重要。组蛋白甲基化的稳态由组蛋白甲基转移酶和去甲基化酶的活性维持平衡。尽管这些甲基转移酶和去甲基化酶是潜在药物发现的合理靶点，但甲基转移酶和去甲基化酶在复杂生物刺激下的活性调节也很重要且尚未明确。为了在生物系统中操纵和研究组蛋白甲基化，我们利用一种表型分析系统筛选了一个生物多样性化合物集（BDCS），该系统可直接测量细胞中的组蛋白3赖氨酸27三甲基化（H3K27me3）水平。BDCS是一组独特的带有靶点注释的化学探针，总共包含5853种化合物，针对736种独特蛋白质，每个靶点有多种最大选择性化合物。在筛选中鉴定出了许多靶点，每个靶点都有多个命中结果。这使我们相信这些靶点和信号通路可能是相关的，包括鉴定非甲基转移酶/去甲基化酶靶点作为H3K27me3的潜在上游调节因子。我们的研究表明，系统设计的化学探针库与表型筛选相结合时可作为强大的药物发现工具。利用这些发现进行的后续研究可能会揭示H3K27me3调控的新的治疗有用途径和靶点。
