High Throughput Screen for Inhibitors of the YEATS2 Histone Acylation Reader

项目摘要 肺癌是美国和世界范围内男性和女性癌症相关死亡的主要原因。 全世界每年有超过40万人死于肺癌，其中肺鳞状细胞癌是最常见的肺癌之一。 肺癌（LUSC）约占30%。然而，LUSC没有有效的治疗方法。LUSC是 一种以多种DNA改变为特征的非小细胞肺癌（NSCLC），包括 3q 26染色体片段的频繁扩增。3q 26片段值得注意，因为它包含 含有YEATS结构域2（YEATS 2）基因，一种在许多人中经常扩增的基因， 癌症，包括LUSC（~50%）、卵巢癌（28%）、头颈癌（25%）和食道癌（25%）。高 YEATS 2 mRNA表达与NSCLC患者的不良预后相关，表明YEATS 2 可能有促进肿瘤的作用。 YEATS 2是保守的后生动物Ada-Two-A-Containing（ATAC）复合物的化学计量亚基 组蛋白乙酰转移酶（HAT）复合物。YEATS 2含有进化上保守的YEATS结构域。我们 先前的研究表明，YEATS 2的YEATS结构域作为组蛋白乙酰化和其他功能的阅读器， 组蛋白酰化类型，如巴豆酰化。重要的是，破坏叶芝组蛋白阅读活性 损害YEATS 2和ATAC复合物的正常功能，导致组蛋白乙酰化降低， 降低靶基因表达，抑制NSCLC细胞生长和存活。这些数据 证明YEATS 2的YEATS结构域是一个潜在的药物靶点，靶向YEATS 2可能 提供治疗NSCLC和以YEATS 2为特征的其他类型癌症的治疗方法 放大 本研究的目的是开发针对组蛋白酰化结合的有效和特异性抑制剂 YEATS 2的YEATS结构域的活性。为此，我们将（1）进行高通量筛选， YEATS 2小分子抑制剂，和（2）在体外和基于细胞的细胞内评估和表征命中化合物。 测定。通过所提出的研究，我们期望鉴定有效的、特异性的YEATS 2 YEATS结构域化学物质 不同化学型的探针。这些化合物将为进一步开发小型 用于靶向治疗的分子。同样，研究界将能够使用这些新的抑制剂， 这是了解YEATS 2在人类癌症中的功能和机制的重要工具。

PROJECT SUMMARY Lung cancer is the leading cause of cancer-related deaths in both men and women in the U.S and worldwide. There are more than 400,000 deaths of lung cancer worldwide each year, among which lung squamous cell carcinoma (LUSC) accounts for about 30%. However, no effective treatments for LUSC are available. LUSC is one type of non-small cell lung cancer (NSCLC) characterized by numerous DNA alterations, including frequent amplification of the 3q26 chromosomal segment. The 3q26 segment is noteworthy because it contains the YEATS domain containing 2 (YEATS2) gene, a gene that is frequently amplified in a number of human cancers, including LUSC (~50%), ovarian (28%), head and neck (25%), and esophagus cancers (25%). High YEATS2 mRNA expression is associated with a poor prognosis of NSCLC patients, indicating that YEATS2 may have a tumor-promoting role. YEATS2 is a stoichiometric subunit of the Ada-Two-A-Containing (ATAC) complex, a conserved metazoan histone acetyltransferase (HAT) complex. YEATS2 contains an evolutionally conserved YEATS domain. We previously showed that the YEATS domain of YEATS2 functions as a reader of histone acetylation and other types of histone acylation such as crotonylation. Importantly, disrupting the YEATS histone reading activity impairs the normal functions of YEATS2 and the ATAC complex, resulting in reduced histone acetylation, decreased target gene expression, and inhibition of cell growth and survival of NSCLC. These data demonstrate that the YEATS domain of YEATS2 is a potential drug target, and that targeting YEATS2 may provide a therapeutic approach for treating NSCLC and other types of cancer characterized by YEATS2 amplification. The objective of this proposal is to develop potent and specific inhibitors targeting the histone acylation binding activity of the YEATS domain of YEATS2. For this, we will (1) conduct a high-throughput screen to identify YEATS2 small-molecule inhibitors, and (2) evaluate and characterize hit compounds in in vitro and cell-based assays. Through the proposed studies, we expect to identify potent, specific YEATS2 YEATS domain chemical probes of different chemotypes. These compounds will provide the basis for further development of small molecules for targeted therapies. Likewise, the research community will be able to use these new inhibitors as important tools to understand the functions and mechanisms of YEATS2 in human cancers.