Mutations of Chromatin and its Modifying Machineries in Malignancies

项目摘要 我的实验室的集体研究集中在功能的分子表征上(S)和 与人类癌症相关的表观遗传修饰物和突变的生化特性。 我们的目标是确定这些因子中的突变如何在分子上改变基因模式。 表达导致细胞过度增殖，并利用这些分子途径识别靶向 癌症治疗学。我们在过去七年的详细结构研究由这笔拨款资助 R35CA197569使我们能够确定Set1/Compass的原子结构，并以此为基础 信息，我们已经产生了抑制其活性的药物，现在我们的实验室正在开发 一种在细胞内调节指南针功能的工具化合物，最终用于临床。在其他方面，我们的 对MLI3/COMPASS的作用分析确定了它与组蛋白脱泛素化酶的相互作用 BAP1，在肿瘤中提供增强子和基因表达调控的中心表观遗传平衡 这是由MLL3突变引起的。基于这些信息，我们在西北大学进行了一项临床试验 研究这种平衡在癌症治疗中的表观遗传学作用的医学。我们还生成了BAP1 ASXL1突变所致癌症中可调节BAP1过度激活功能的抑制物 而这些BAP1抑制剂正在我们的实验室作为生化和临床工具进行进一步的研究。我们的 组蛋白H3K27M突变在弥漫性桥脑胶质瘤(DIPG)中的作用 包含Brd2和Brd4等因子的BET结构域在DIPG发病机制中的作用。我们 正在与芝加哥Lurie儿童医院的同事合作，在临床上测试这些发现。这些 研究还发现，地下墓穴是一种类似H3K27M和H3K27M的内源性因子 调节PRC2功能。正如将在申请中描述的，实验室中的其他研究已经探索 不同表观遗传因子在发育调控和癌症中的作用机制。另外， 最近对癌症和自然衰老过程中的体细胞突变的编目发现了大量的 MLL1-4、UTX、Set1A/B和其他表观遗传因子的成分突变。鉴于我们有 在过去的二十年里，我们实验室针对这些因素开发了一套奇妙的试剂和工具， 它们的相关蛋白质、染色质和其他染色质修饰物在多个模型系统中，我的实验室是 在一个非常独特和强有力的位置来确定这些因子参与癌症的分子基础 发病机制，从而达到靶向治疗的目的。此R35续订申请的目标是 是我们对三胸罗盘的完整分子和生化特征的延续 家族和BAP1复合体在发育基因表达调控中的作用 突变与人类癌症的发病机制有关。

Project Summary Collective studies from my laboratory have focused on the molecular characterization of the function(s) and biochemical properties of the epigenetic modifiers and the mutations that are associated with human cancer. Our goal has been to determine how the mutations in these factors molecularly change the pattern of gene expression resulting in cellular hyperproliferation and use these molecular pathways to identify targeted therapeutics for cancer. Our detailed structural studies supported during the past seven years by this grant R35CA197569 have allowed us to identify the atomic structure of Set1/COMPASS, and based on this information, we have generated inhibitors towards its activity that are now being developed in our laboratory as a tool compound to regulate COMPASS's function in cells for the ultimate use in clinic. On other fronts, our analyses of the role of MLL3/COMPASS identified its interactions with the histone deubiquitinating enzyme BAP1, providing a central epigenetic balance at enhancers and the regulation of gene expression in cancer caused as the result of MLL3 mutations. Based on this information, we have a clinical trial at Northwestern Medicine investigating the epigenetic role of this balance in cancer therapy. We have also generated BAP1 inhibitors that can regulate hyperactivated BAP1 function in cancers caused as the result of ASXL1 mutations and such BAP1 inhibitors are being further investigated in our laboratory as biochemical and clinical tools. Our detailed analysis of the role of histone H3K27M mutations in diffuse intrinsic pontine gliomas (DIPG) resulted in the identification of a role for BET-domain containing factors such as Brd2 and Brd4 in DIPG pathogenesis. We are collaborating with colleagues at Lurie Children's Hospital of Chicago to test these findings in clinic. These studies also resulted in the identification of CATACOMB as an endogenous factor mimicking H3K27M and regulating PRC2 function. As will be described in the application, other studies in the laboratory have probed the mechanistic properties of diverse epigenetic factors in developmental regulation and in cancer. Additionally, recent cataloging of somatic mutations in cancer and during natural aging has identified a large number of mutations in the components of the MLL1-4, UTX, Set1A/B, and other epigenetic factors. Given that we have developed a fantastic set of reagents and tools over the past twenty years in our laboratory towards these factors, their associated proteins, chromatin, and other chromatin modifiers in multiple model systems, my laboratory is in a very unique and strong position to define the molecular bases of these factors’ involvement in cancer pathogenesis, and thus, for the purpose of targeted therapeutics. The goals of this R35 renewal application are the continuation of our full molecular and biochemical characterization of the trithorax COMPASS family and BAP1 complexes in the regulation of developmental gene expression, and how their mutations contribute to the pathogenesis of human cancer.