Targeting epigenetic reader GAS41

摘要 组蛋白的翻译后修饰在调节染色质转录中起着至关重要的作用 在人类癌症中。组蛋白乙酰化与活跃的基因转录有关，并在 肿瘤发生学。事实上，以组蛋白修饰调控中的蛋白质为靶标的小分子 被开发为一种非常有前途的抗癌药物，目前有一些化合物正在进行临床试验 (例如，BRD4抑制剂)。最近，叶芝结构域被发现是一种新的乙酰组蛋白阅读器 域名。最初发现GAS41在23%的胶质母细胞瘤和80%的星形细胞瘤中扩增。新兴 研究强烈表明GAS41是非小细胞肺癌(NSCLC)的癌基因。GAS41是 在非小细胞肺癌中频繁扩增和GAS41的敲除或与乙酰化的相互作用中断 组蛋白抑制肺癌细胞生长。 我们最近发现GAS41是双乙酰化H3组蛋白的阅读器。全长GAS41是二聚体 细胞，并以高亲和力结合二乙酰化的H3。基于这一发现，我们开发了生化套件 适于表征GAS41蛋白-蛋白质相互作用和小分子鉴定的方法 抑制剂。在这项提案中，我们计划利用高通量开发GAS41的小分子抑制剂 密歇根大学CCG的筛查(HTS)。GAS41的小分子抑制剂将得到验证 并通过一系列生化和生物物理实验进行表征。活性最强的 化合物将在基于细胞的分析中进行表征，以评估GAS41与 并了解染色质的作用机制。选定的GAS41抑制剂也将在 肺癌细胞株。综上所述，我们希望能够识别出具有很高价值的化学探针化合物 适合机理研究的GAS41蛋白质-蛋白质相互作用为发展GAS41蛋白质铺平道路 有效的体内活性GAS41抑制剂。

Abstract Post-translational modifications on histone proteins play an essential role in regulating chromatin transcription in human cancers. Histone acetylation is associated with active gene transcription and plays a crucial role in tumorigenesis. Indeed, small molecules targeting proteins involved in regulation of histone modifications are being explored as a very promising anti-cancer agents, with a number of compounds currently in clinical trials (e.g. BRD4 inhibitors). Recently, the YEATS domains have been discovered as novel acetyl-histone reader domains. GAS41 was originally found to be amplified in 23% glioblastomas and 80% astrocytoma. Emerging studies strongly implicate GAS41 as an oncogene in Non-Small Cell Lung Cancer (NSCLC). GAS41 is frequently amplified in NSCLC and knockdown of GAS41 or disruption of the interaction with acetylated histones suppresses lung cancer cell growth. We have recently found that GAS41 is a reader of di-acetylated H3 histone. Full-length GAS41 is dimeric in cells and binds di-acetylated H3 with high affinity. Based on this finding we developed suites of biochemical assays suitable for characterization of GAS41 protein-protein interactions and identification of small molecule inhibitors. In this proposal we plan to develop small molecule inhibitors of GAS41 using high throughput screening (HTS) in CCG at the University of Michigan. Small molecule inhibitors of GAS41 will be validated and characterized in a series of biochemical and biophysical experiments. Activity of the most potent compounds will be characterized in cell-based assays to assess the disruption of GAS41 interactions with chromatin and understand mechanism of action. Selected GAS41 inhibitors will be also profiled in a panel of lung cancer cell lines. In summary, we expect to identify highly valuable chemical probe compounds targeting GAS41 protein-protein interactions suitable for mechanistic studies and pave the way towards development of potent in vivo active GAS41 inhibitors.