The function of Topoisomerase I SUMOylation in transcription and chemoresistance

拓扑异构酶 I SUMO 化在转录和化疗耐药中的作用

• 批准号: 10132345
• 负责人: Yilun Liu
• 金额: $ 34.6万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10132345
• 关键词: Acute; Angelman Syndrome; Biochemical; Cell Culture Techniques; Cell Death; Cell Line; Cell physiology; Cells; Chemoresistance; Chemotherapy-Oncologic Procedure; Chromatin; Coupling; DNA; DNA Damage; DNA Repair; Disease; Enzymes; Event; Excision; Exhibits; Exons; FDA approved; Gene Activation; Gene Expression; Genetic Transcription; Genome Stability; High-Throughput DNA Sequencing; Human; Infection; Inflammation; Inflammatory Response; Introns; Link; Lysine; Malignant Neoplasms; Maps; Mediating; Messenger RNA; Modeling; Modification; Molecular; Movement; Pathogenesis; Pathogenicity; Pathway interactions; Pattern; Pharmaceutical Preparations; Play; Poison; Positioning Attribute; RNA; RNA Polymerase II; RNA Processing; RNA Splicing; Reaction; Regulation; Research; Resistance; Role; Sepsis; Sepsis Syndrome; Source; Specificity; Spliced Genes; Spliceosomes; Sumoylation Pathway; Superhelical DNA; Survival Rate; Testing; Therapeutic; Topoisomerase; Topoisomerase I inhibition; Topoisomerase-I Inhibitor; Topotecan; Transcription Initiation Site; Transcriptional Activation; Transcriptional Regulation; Type I DNA Topoisomerases; Variant; autism spectrum disorder; base; cancer cell; cancer therapy; carcinogenicity; cell killing; chromatin immunoprecipitation; cytotoxic; cytotoxicity; drug candidate; drug development; genome-wide; inhibitor/antagonist; innovation; insight; mutant; novel; novel strategies; novel therapeutics; prevent; promoter; protein protein interaction; recruit; therapeutic candidate; therapeutic target; tissue culture; tool; transcriptome sequencing; treatment strategy; virtual

Project Summary Topoisomerase I (TOP1) is a FDA-approved therapeutic target for late-stage cancers with a recently expanded therapeutic potential. Canonical TOP1 poisons, such as topotecan, kill rapidly dividing cells by preventing the release of TOP1 from DNA during its topoisomerase reaction to relax supercoiled DNA, leading to DNA breaks. However, it was recently discovered that TOP1 poisons also alter gene expression linked to autism spectrum disorders (ASD) and acute inflammatory response, among others, via a mechanism that is independent of DNA damage. Indeed, in addition to relaxing supercoiled DNA, TOP1 also regulates mRNA splicing and gene expression through its interaction with RNA polymerase II (RNAPII) and splicing factors. Therefore, new drugs that modulate the transcriptional function of TOP1 without causing DNA damage could greatly increase the therapeutic scope of TOP1, and provide treatment options for difficult diseases. However, the mechanisms linking TOP1 to mRNA regulation are not well established. Our recent discovery that SUMOylation of the lysine (K) residues 391 and 436 of TOP1 regulates its transcriptional function provides a crucial insight that will accelerate mechanistic advances. Due to this new insight, we are well-positioned to uncover the detailed molecular mechanism linking TOP1 and mRNA regulation. This proposal will first test a novel hypothesis that TOP1 K391/K436 SUMOylation regulates RNAPII movement and the coupling of RNAPII and spliceosome to facilitate gene transcription (Aim 1). We will evaluate the transcriptional landscape and splicing patterns, including the presence of splice variants that are dependent on TOP1 K391/K436 SUMOylation. We will identify additional cellular processes that may be altered by the inhibition of the TOP1 transcriptional function to uncover its full potential for new disease treatments (Aim 2). We will also develop a novel molecular strategy to disrupt the effects of TOP1 on mRNA regulation. This novel strategy will facilitate drug development for a wide range of disorders, including inflammation-induced sepsis and ASD. Finally, in addition to facilitating transcription, K391/K436 SUMOylation also suppresses TOP1 topoisomerase activity. Because the sensitivity to TOP1 poisons positively correlates with the level of TOP1 topoisomerase activity in cells, we will test an exciting possibility that transiently blocking TOP1 SUMOylation by this SUMOylation inhibitor could hypersensitize cells to TOP1 poisons during cancer chemotherapy (Aim 3).

项目摘要 拓扑异构酶I（TOP1）是FDA批准的治疗靶标，用于近期扩展的后期癌症 治疗潜力。典型的Top1毒物（例如拓扑克），通过防止该细胞杀死细胞 在拓扑异构酶酶反应中从DNA释放TOP1以放松超涂DNA，导致DNA 休息。但是，最近发现Top1毒物也改变了与自闭症相关的基因表达 频谱障碍（ASD）和急性炎症反应，以及其他机制 独立于DNA损伤。实际上，除了放松超螺旋DNA外，Top1还调节mRNA 剪接和基因表达通过与RNA聚合酶II（RNAPII）和剪接因子的相互作用。 因此，调节TOP1转录功能而不会引起DNA损伤的新药物可能 大大增加了TOP1的治疗范围，并为困难疾病提供治疗选择。然而， 将TOP1与mRNA调控联系起来的机制尚未确定。我们最近发现 TOP1的赖氨酸（K）残基391和436的Sumoylation调节其转录函数提供了 至关重要的见解将加速机理的进步。由于这个新见解，我们对 发现连接TOP1和mRNA调控的详细分子机制。该建议将首先测试 Top1 K391/K436 Sumoylation调节RNAPII运动和RNAPII耦合的新型假设 和剪接体以促进基因转录（AIM 1）。我们将评估转录格局和 剪接模式，包括依赖TOP1 K391/K436的剪接变体的存在 sumoylation。我们将确定可能通过抑制TOP1改变的其他细胞过程 转录功能可揭示其对新疾病治疗的全部潜力（AIM 2）。我们还将开发一个 新的分子策略破坏了TOP1对mRNA调节的影响。这种新颖的策略将有助于 多种疾病的药物开发，包括炎症引起的败血症和ASD。最后，在 除了促进转录外，K391/K436 Sumoylation还抑制了TOP1拓扑异构酶活性。 因为对Top1毒物的敏感性与TOP1拓扑异构酶活性的水平正相关 单元格，我们将测试一种令人兴奋的可能性，该可能性通过这种sumoylation瞬时阻止top1 sumoylation 在癌症化学疗法期间，抑制剂可以使细胞超敏于TOP1毒物（AIM 3）。