Targeting crotonyl-lysine chromatin readers to disrupt pathogenic gene expression in leukemia

项目总结/摘要 支持细胞不受抑制的增殖需求，并通过以下方式维持恶性细胞状态： 在细胞分裂的连续循环中，癌细胞依赖于谱系特异性的失调基因表达， 程序.因此，作为染色质依赖性转录控制的基础的染色质调节因子已经被研究。 被认为是破坏或减少致病性转录信号传导的有吸引力的靶点。在此之前，我领导了一个 研究工作，建立ENL，转录辅激活因子和染色质阅读蛋白，作为一个关键的 急性白血病在细胞和动物模型系统中存活的必要条件。值得注意的是，它的YEATS域， 巴豆酰赖氨酸结合染色质阅读器结构域，其将ENL锚定在转录活性启动子处， 基因，是白血病中ENL的促增殖作用所必需的。此外，初步数据显示， KAT 6A，一种组蛋白乙酰转移酶和巴豆酰赖氨酸染色质阅读蛋白，也是急性心肌梗死所必需的。 白血病发病机制在这里，我建议进一步考虑巴豆酰赖氨酸阅读器蛋白作为潜在的 急性白血病的治疗机会，有以下三个具体目标。在第一个目标中，我将决定 细胞通路和辅助因子调节急性白血病对ENL丧失的反应和抵抗。 在此目标下提出的遗传实验将确定调节ENL的细胞途径和辅因子 靶向生物学以了解其促进白血病细胞增殖和存活的作用机制。在 第二个目标，我将测试ENL YEATS结构域可以被小分子抑制剂破坏的假设， 其将作为ENL蛋白功能的化学探针和翻译的起始点， ENL靶向白血病治疗的发展。我将应用高通量化学筛选，基于结构 药物设计和迭代药物化学，以发现和优化一流的ENL YEATS抑制剂。在 第三个目标，我将评估KAT 6A识别巴豆酰赖氨酸在白血病发病机制中的作用;我将使用 遗传方法和靶向蛋白质降解，以确定巴豆酰赖氨酸是否被 KAT 6A的双PHD指结构域（DPF）是急性白血病存活所必需的。每一个拟议的目标运作 在早期阶段的药物发现工作（即目标验证和化学工具发现）中，如果成功，将 提供生物学见解和化学工具， 分子靶向抗癌药物。遵循学术药物开放式创新模式 发现，所有的化学探针，生物工具，和基因组数据集来自这项工作将作出 公开和免费提供，不限制使用，以扩大和加速这项工作的影响。

PROJECT SUMMARY/ABSTRACT To support the cellular demands of unchecked proliferation and to maintain a malignant cell state through successive rounds of cellular division, cancer cells depend on lineage-specific, dysregulated gene expression programs. Thus, the chromatin regulators that underlie chromatin-dependent transcription control have been considered attractive targets to disrupt or diminish pathogenic transcriptional signaling. Previously, I led a research effort that established ENL, a transcriptional co-activator and chromatin reader protein, as a critical requirement for the survival of acute leukemia in cellular and animal model systems. Notably, its YEATS domain, a crotonyl-lysine-binding chromatin reader domain that anchors ENL at the promoters of transcriptionally active genes, is required for the pro-proliferative effects of ENL in leukemia. Moreover, preliminary data suggests that KAT6A, a histone acetyltransferase and crotonyl-lysine chromatin reader protein, is also required for acute leukemia pathogenesis. Here, I propose to further consider crotonyl-lysine reader proteins as potential therapeutic opportunities in acute leukemia with the following three specific aims. In the first aim, I will determine the cellular pathways and co-factors regulating response and resistance to loss of ENL in acute leukemia. Genetic experiments proposed within this aim will determine the cellular pathways and co-factors regulating ENL target biology to understand its mechanism of action in promoting leukemia cell proliferation and survival. In the second aim, I will test the hypothesis that the ENL YEATS domain can be disrupted by small-molecule inhibitors, which will serve both as chemical probes of ENL protein function and as starting points for translational development of ENL-targeted leukemia therapy. I will apply high-throughput chemical screens, structure-based drug design, and iterative medicinal chemistry to discover and optimize first-in-class ENL YEATS inhibitors. In the third aim, I will assess the role of crotonyl-lysine recognition by KAT6A in leukemia pathogenesis; I will use genetic approaches and targeted protein degradation to determine whether crotonyl-lysine recognition by the double PHD finger domain (DPF) of KAT6A is required for acute leukemia survival. Each proposed aim operates within early-stage drug discovery efforts (i.e. target validation and chemical tool discovery) and if successful, will provide biological insights and chemical tools that will enable timely translational development of novel, molecularly-targeted cancer medicines. In keeping with the model of open-innovation in academic drug discovery, all chemical probes, biological tools, and genomic datasets derived from this work will be made publicly and freely available, without restriction on use, so to broaden and accelerate the impact of this work.