Inhibitor Reprogramming of a Human Histone Deacetylase Protein Interaction Network

 描述(由申请人提供)： 组蛋白脱乙酰酶(HDAC)在癌症中起着重要的作用，FDA批准的HDAC抑制剂Suberoylanilide草酮酸(SAHA)用于治疗进展性、持续性或复发性疾病患者的晚期和难治性皮肤T细胞淋巴瘤。SAHA是225多项临床试验的一部分。抑制HDAC活性会导致核心组蛋白和其他蛋白质的乙酰化改变，从而导致基因表达的变化，导致细胞生长停滞和细胞凋亡。然而，尽管美国在萨哈作为人类疾病治疗药物的研究上投入了大量资金，但人们经常说，萨哈的作用机制仍然知之甚少。我们实验室最近的发现表明，SAHA和HDAC抑制剂实际上对HDAC复合体具有更广泛的非催化作用。在这里，我们将使用创新的生化、定量蛋白质组学、计算和基因组学技术来进一步阐明SAHA的作用机制。特别是，我们将分析SAHA诱导的人类HDAC蛋白相互作用网络的破坏，并确定通过破坏这个网络可以解释SAHA诱导的基因表达变化的哪一部分。在具体目标1中，我们将完成蛋白质相互作用网络的组装，并对该网络中存在的多个复合体进行生化剖析。此外，我们将开发新的计算方法来组装蛋白质相互作用网络，并确定该网络中相互作用的强度。在具体目标2中，我们将完成对SAHA诱导的网络动力学的分析，并剖析网络中断导致的基因表达变化。在第三个特定目标中，我们将对一个乳腺癌细胞株进行有针对性的研究，并将这些结果与特定目标1和2中使用的标准细胞系进行比较。通过比较和对比这些结果，我们将确定SAHA对这个人类HDAC网络的细胞特异性作用。完成本申请后，我们将对多种类型的细胞有详细的了解 SAHA对组蛋白相互作用网络的影响以及该网络的破坏所导致的基因表达变化。这些信息将产生对 阐明SAHA的作用机理，促进新型HDAC抑制剂的开发。

 DESCRIPTION (provided by applicant): Histone deacetylases (HDACs) play important roles in cancer, and the FDA approved HDAC inhibitor, suberoylanilide hydroxamic acid (SAHA), is used for the treatment of advanced and refractory cutaneous T-cell lymphoma in patients with progressive, persistent, or recurrent disease. SAHA is part of more than 225 clinical trials. Inhibition of HDAC activity results in altered acetylation of core histones and other proteins that cause changes in gene expression, resulting in cell growth arrest and apoptosis. However, despite the massive investment the United States has made into the study of SAHA as a drug for human disease treatment, it is routinely stated that the mechanism of action of SAHA remains poorly understood. Recent findings from our lab suggest that SAHA and HDAC inhibitors actually have broader, noncatalytic effects on HDAC complexes. Here we will use innovative biochemical, quantitative proteomic, computational, and genomic technologies to further elucidate the mechanism of action of SAHA. In particular, we will analyze the SAHA induced disruption of a human HDAC protein interaction network and determine which fraction of SAHA induced gene expression changes can be accounted for by disrupting this network. In specific aim 1 we will complete our assembly of the protein interaction network and biochemically dissect the multiple complexes present in this network. Furthermore, we will develop novel computational approaches for protein interaction network assembly and determine the strength of interactions in this network. In specific aim two, we will finalize our analysis of the SAHA induced dynamics of the network and dissect the gene expression changes resulting from network disruption. In the third specific aim, we will carry out focused studies on a breast cancer cell line and compare these results to the standard cell line used in specific aims 1 and 2. By comparing and contrasting these results, we will determine the cell specific effects of SAHA on this human HDAC network. Upon completion of this application, we will have a detailed understanding in multiple cell types of the effects of SAHA on a histone protein interaction network and the resulting gene expression changes mediated by the disruption of this network. This information will yield novel insights into the mechanism of action of SAHA and facilitate future development of novel HDAC inhibitors.