Identification and characterization of selective azaphilone inhibitors of HuR-mRNA interactions

HuR-mRNA 相互作用的选择性阿扎菲酮抑制剂的鉴定和表征

• 批准号: 10672205
• 负责人: Tessa Epstein
• 金额: $ 3.58万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-06 至 2024-08-05
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10672205
• 关键词: 3' Untranslated Regions; Adenine; Binding; Biochemical Reaction; Biological; Biological Assay; Cell model; Cell physiology; Cells; Chemicals; Colon; Colon Carcinoma; Complex; Cytoplasm; Development; Disease; Disease model; Elements; Functional disorder; Gene Expression; Gene Expression Regulation; Generations; Goals; Half-Life; Health; HuR protein; Human; In Vitro; Libraries; Link; Location; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Measures; Messenger RNA; Methods; Natural Products; Nature; Neoplasm Metastasis; Oncogenic; Outcome; Pancreas; Pathway interactions; Process; Proteins; RNA; RNA Recognition Motif; RNA Splicing; RNA-Binding Proteins; Reaction; Regulation; Research; Role; Route; Therapeutic; Translations; United States; Up-Regulation; Uridine; Work; analog; cancer cell; drug discovery; effective therapy; functional outcomes; inhibitor; innovation; mRNA Stability; novel; novel therapeutics; nucleocytoplasmic transport; overexpression; scaffold; scale up; screening; small molecule; small molecule inhibitor; targeted treatment; therapeutic development; tumor; tumor growth; tumorigenesis

PROPOSAL SUMMARY RNA-binding proteins (RBPs) regulate gene expression through binding to mRNAs, thus influencing rates of translation, mRNA subcellular location, and mRNA half-life. The specific functional outcome of an RBP-mRNA interaction is dependent on the identity of both binding partners. However, it is challenging to predict the functional consequences of a specific interaction due to the diverse combinations of RBP-mRNA interactions that occur within the cell. The RBP HuR binds has a multitude of different mRNA binding partners, allowing HuR to control many critical cellular functions. Dysregulation of the HuR-mRNA interaction network is notably implicated in cancers such as colon, lung, and pancreatic cancers, the three most deadly cancers in the United States. A recently discovered azaphilone HuR inhibitor is very potent but lacks the selectivity required for therapeutic development or to dissect the complex network of HuR mRNA binding partners. Traditional synthetic methods are a major roadblock towards enantioselective azaphilone synthesis, making it infeasible to screen analogs for more favorable bioactivity. However, recent innovation has produced a straightforward, one-pot biocatalytic route that can be used to generate large, diverse azaphilone libraries. I aim to identify novel azaphilones that act as potent and selective HuR-mRNA inhibitors using a combined biocatalytic generation and HuR binding assay platform. I will characterize and validate HuR-azaphilone interactions and identify selective azaphilone inhibitors of HuR both in vitro and in cellulo. Next, I will investigate functional implications of disrupting specific HuR-mRNA interactions in a cancer cell model. Completion of the proposed work will result in the identification and characterization of novel azaphilones that are potent and selective HuR-mRNA interactions and can be further developed as chemical probes and cancer therapeutics.

提案摘要 RNA结合蛋白（RBP）通过与mRNA结合调节基因表达，从而影响了速率 翻译，mRNA亚细胞位置和mRNA半衰期。 RBP-MRNA的特定功能结果 相互作用取决于两个结合伙伴的身份。但是，预测 由于RBP-MRNA相互作用的不同组合，特定相互作用的功能后果 这发生在细胞内。 RBP HUR BINDS具有多种不同的mRNA绑定伙伴，允许HUR 控制许多关键细胞功能。 HUR-MRNA相互作用网络的失调尤其是 与结肠，肺和胰腺癌等癌症有关，这三种致命的癌症是曼联的三种癌症 国家。最近发现的Azaphilone hur抑制剂非常有效，但缺乏选择性 治疗开发或剖析HUR mRNA结合伙伴的复杂网络。传统的合成 方法是通向对映选择性azaphilone合成的主要障碍，使其无法筛选 类似于更有利的生物活性。但是，最近的创新产生了一个简单的一锅 可用于生成大型的，多样化的Azaphilone库的生物催化途径。我旨在识别小说 使用合并的生物催化产生和 HUR绑定测定平台。我将表征和验证hur-azaphilone互动并确定选择性 HUR在体外和纤维素中均具有Azaphilone抑制剂。接下来，我将调查破坏的功能含义 癌细胞模型中的特定hur-mRNA相互作用。拟议工作的完成将导致 具有有效和选择性的HUR-MRNA相互作用的新型Azaphilones的识别和表征 并且可以作为化学探针和癌症治疗剂进一步开发。