Characterization of the SIN3A and SIN3B HDAC complexes

项目摘要 SAHA是FDA批准的化疗药物，已成功用于治疗T-T- 细胞性淋巴瘤。这种组蛋白脱乙酰酶(HDAC)抑制剂是过去或正在进行的240多种药物中的一部分 临床试验(Clinicaltrials.gov)，旨在检查SAHA作为许多其他疾病的治疗选择的有效性 癌症亚型和病理学。尽管在临床层面上进行了广泛的研究，但治疗 SAHA和其他HDAC抑制剂的潜力受到我们对 调节它们活动的分子机制。我们建议的研究将揭示这些问题 SIN3-HDAC络合物对SAHA的响应机理和表征。我们的目标是 表征SIN3A和SIN3B络合物的功能动力学。我们假设SIN3A和 SIN3B复合体具有不同的功能属性，并且对HDAC有不同的响应 抑制剂。我们的目标将通过两个目标来实现。在目标1中，我们将描述 SIN3A和SIN3B络合物的生物物理性质。研究SIN3A和SIN3B核心复合体 架构安排和对HDAC抑制剂的响应，我们将使用交联体 光谱以确定复合体中蛋白质-蛋白质相互作用的位置并监测其 对HDAC抑制剂的反应。在目标2中，我们将考察SIN3A和SIN3B所扮演的独特角色 SH-SY5Y神经母细胞瘤和HEK293细胞内的蛋白质及其对细胞周期的影响 生物途径。在解决这些核心生物问题的过程中，我们将使 为定量蛋白质组学领域做出的重大贡献 通过交联质谱定量蛋白质-蛋白质相互作用的动力学。作为萨哈 和其他HDAC抑制剂已经是临床治疗计划的组成部分，拟议的研究将 产生将立即有用的信息，因为我们定义了SAHA的治疗潜力。长- 长期而言，这些发现将为开发靶向HDAC抑制剂提供基础 HDAC复合体的特定方面，几乎不会产生偏离目标的效果。

Project Summary SAHA is an FDA-approved chemotherapeutic agent that has found success as a treatment for T- cell lymphoma. This histone deacetylase (HDAC) inhibitor is additionally part of over 240 past or ongoing clinical trials (clinicaltrials.gov) that seek to examine SAHA efficacy as a treatment option for many other cancer subtypes and pathologies. Despite widespread study at the clinical level, the therapeutic potential of SAHA and other HDAC inhibitors is hampered by our rudimentary understanding of the molecular mechanisms that mediate their actions. Our proposed studies will shed light upon such mechanisms and characterize the response of the SIN3 HDAC complexes to SAHA. Our objective is to characterize the functional dynamics of SIN3A and SIN3B complexes. We hypothesize that SIN3A and SIN3B complexes possess divergent functional attributes and are differentially responsive to HDAC inhibitors. Our objective will be addressed through two aims. In aim 1, we will characterize the biophysical properties of SIN3A and SIN3B complexes. To examine the SIN3A and SIN3B core complex architectural arrangements and responses to HDAC inhibitors, we will employ crosslinking mass spectrometry to identify sites of protein-protein interactions within the complex and monitor their responses to HDAC inhibitors. In aim 2, we will examine the unique roles held by SIN3A and SIN3B proteins within SH-SY5Y neuroblastoma and HEK293 cells and characterize their influences on biological pathways. In the process of addressing these central biological issues, we will make significant contributions to the field of quantitative proteomics by developing an assay system capable of quantifying the dynamics of protein-protein interactions via crosslinking mass spectrometry. As SAHA and other HDAC inhibitors are already components of clinical treatment plans, the proposed studies will produce information that will be immediately useful as we define the therapeutic potential of SAHA. Long- term, these findings will provide the groundwork for the development of HDAC inhibitors that target specific aspects of HDAC complexes and produce few off-target effects.