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

基本信息

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
项目类别：
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

项目摘要

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).

项目概要拓扑异构酶 I (TOP1) 是 FDA 批准的晚期癌症治疗靶点，最近扩大了治疗范围治疗潜力。典型的 TOP1 毒物，例如托泊替康，通过阻止细胞分裂来杀死快速分裂的细胞。在拓扑异构酶反应期间从 DNA 中释放 TOP1，以松弛超螺旋 DNA，从而产生 DNA 休息。然而，最近发现 TOP1 毒物也会改变与自闭症相关的基因表达谱系障碍（ASD）和急性炎症反应等，通过一种机制与 DNA 损伤无关。事实上，除了松弛超螺旋 DNA 之外，TOP1 还调节 mRNA 通过与 RNA 聚合酶 II (RNAPII) 和剪接因子的相互作用来实现剪接和基因表达。因此，调节 TOP1 转录功能而不引起 DNA 损伤的新药可能大大增加了TOP1的治疗范围，为疑难疾病提供了治疗选择。然而，将 TOP1 与 mRNA 调节联系起来的机制尚未明确。我们最近的发现是 TOP1 赖氨酸 (K) 残基 391 和 436 的 SUMO 化调节其转录功能提供了将加速机械进步的重要见解。由于这一新的见解，我们处于有利地位揭示连接 TOP1 和 mRNA 调控的详细分子机制。该提案将首先测试 TOP1 K391/K436 SUMOylation 调节 RNAPII 运动和 RNAPII 偶联的新假设和剪接体以促进基因转录（目标 1）。我们将评估转录景观并剪接模式，包括依赖于 TOP1 K391/K436 的剪接变体的存在 SUMO化。我们将确定可能因抑制 TOP1 而改变的其他细胞过程转录功能以揭示其新疾病治疗的全部潜力（目标 2）。我们还将开发一个破坏 TOP1 对 mRNA 调节影响的新分子策略。这一新颖的策略将有利于针对多种疾病的药物开发，包括炎症引起的败血症和 ASD。最后，在除了促进转录之外，K391/K436 SUMO 化还抑制 TOP1 拓扑异构酶活性。因为对TOP1毒物的敏感性与TOP1拓扑异构酶活性水平呈正相关。细胞，我们将测试一个令人兴奋的可能性，即通过这种 SUMOylation 暂时阻断 TOP1 SUMOylation 抑制剂可以在癌症化疗期间使细胞对 TOP1 毒物过敏（目标 3）。