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: 10536274
• 负责人: Tessa Epstein
• 金额: $ 3.48万
• 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/10536274
• 关键词: 3' Untranslated Regions; Adenine; Binding; Biochemical Reaction; Biological; Biological Assay; Cell model; Cell physiology; Cells; Chemicals; Colon; Colon Carcinoma; Complex; 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; base; 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结合来调节基因表达，从而影响基因表达的速率。 翻译、mRNA亚细胞定位和mRNA半衰期。RBP-mRNA的特定功能结果 相互作用取决于两个结合配偶体的身份。然而，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 发生在细胞内。RBP HuR结合有多种不同的mRNA结合伴侣，使得HuR 来控制许多关键的细胞功能。HuR-mRNA相互作用网络的失调是显著的。 与结肠癌、肺癌和胰腺癌等癌症有关，这是美国三种最致命的癌症。 States.最近发现的氮杂菲酮HuR抑制剂是非常有效的，但缺乏所需的选择性， 治疗开发或剖析HuR mRNA结合伴侣的复杂网络。传统合成 方法是对映选择性合成azaphilone的主要障碍，使得筛选不可行。 更有利的生物活性的类似物。然而，最近的创新产生了一个简单的，一锅 生物催化途径，可用于产生大的，多样化的azaphilone文库。我的目标是确定小说 使用组合的生物催化生成作为有效和选择性HuR-mRNA抑制剂的氮杂菲酮， HuR结合测定平台。我将描述和验证HuR-阿扎菲隆相互作用，并确定选择性 在体外和在细胞中的HuR的azaphilone抑制剂。接下来，我将研究干扰的功能含义， 在癌细胞模型中特异性HuR-mRNA相互作用。完成拟议工作后， 作为有效和选择性HuR-mRNA相互作用的新型氮杂菲酮的鉴定和表征 并可进一步开发为化学探针和癌症治疗剂。