Development of Location-specific Sialidase Inhibitors

位置特异性唾液酸酶抑制剂的开发

• 批准号: 10359898
• 负责人: XUE-LONG SUN
• 金额: $ 44.55万
• 依托单位: CLEVELAND STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10359898
• 关键词: Active Sites; Acute; Affect; Binding Sites; Biological; Biological Process; Cell membrane; Cell surface; Cells; Chronic; Dangerousness; Data Analyses; Development; Disease; Education; Enzymes; Excision; Family; Glycolipids; Glycoproteins; Glycoside Hydrolases; Immune; Inflammatory; Interleukin-6; Investigation; Learning; Location; Lysosomes; Measures; Mediating; Membrane; Methods; Modernization; Molecular; Neuraminidase; Pathologic; Pathway interactions; Pharmaceutical Preparations; Pharmacological Treatment; Physiological; Polysaccharides; Production; Receptor Activation; Receptor Signaling; Regulation; Reporting; Research; Research Infrastructure; Sepsis; Sialic Acids; Signal Pathway; Signal Transduction; Substrate Specificity; TLR4 gene; TNF gene; Techniques; Testing; Toll-like receptors; Universities; Vertebrates; Western Blotting; analog; cell type; cytokine; design; enzyme activity; in vivo; inhibitor/antagonist; innovation; macrophage; microbial; mitochondrial membrane; neglect; novel; novel strategies; novel therapeutic intervention; receptor; skills; tool; undergraduate student

Project Summary/Abstract Sialidases (also called neuraminidases) are glycosidases responsible for the removal of sialic acid (Sia) residues (desialylation) from glycan portions of glycocojugates. By desialylation, sialidases are able to modulate the functionality and stability of the Sia-containing molecules and are involved in both physiological and pathological pathways. Previous and our recent study indicate that lysosomal Neu1 sialidase could relocate to the cell surface of macrophages upon LPS stimulation, where it causes desialylation of TLR4 receptor, leading to TLR4 activation and subsequent production of pro-inflammatory cytokines. Dysregulation of TLR4 activation by LPS is responsible for chronic and acute inflammatory disorders that often causes dangerous disease like sepsis that still lacks specific pharmacological treatment. The first objective of this application is to quantitatively profile the location-specific expression of Neu1 sialidase that is critical for TLR4 activation and its subsequent signal transduction. Sialidase inhibitors are useful tools for studying sialidase function and related mechanisms of the biological pathways. More importantly, effective sialidase inhibitors can be used as drugs to regulate the pathological pathways caused by sialidase, such as dysregulated TLR4 activation. Our recent study indicates that currently available pan sialidase inhibitor and microbial sialidase inhibitors could not inhibit mammalian sialidase effectively. Several mammalian sialidase inhibitors have been reported. However, current sialidase inhibitor design has usually focused on active-site binding, neglecting the subcellular localization of the active enzyme, therefore, they are less effective in vivo or may be even toxic as they will affect other sialidases inside of the cells. The second objective of this application is to develop location-specific inhibitor for Neu1 sialidase and define the Neu1 sialidase’s involvement in LPS/TLR4 signaling pathway. The objectives of this project will be accomplished by three specific aims: (1) Profile Neu1 sialidase expression and cell surface relocation in macrophages upon LPS stimulation; (2) Develop lysosome-targeting Neu1 sialidase inhibitors for effective regulating desialylation in LPS/TLR4 signaling pathway; (3) Develop cell surface-targeting Neu1 sialidase inhibitors for effective regulating desialylation in LPS/TLR4 signaling pathway. This study is innovative because it uses a novel approach that overcomes the current limitations in (a) profiling sialidase expression and relocation and (b) inhibiting sialidase at subcellular location. The proposed project is significant because it will (i) uncover specific desialylation that is critical to the LPS/TLR4 signal pathway and (ii) develop novel sialidase inhibitors for effective regulation of desialylation in LPS/TLR4 signaling pathway. Finally, this proposal will enhance the infrastructure of research and education at Cleveland State University, allowing undergraduate students to learn a broad spectrum of experimental techniques, data analysis and presentation skills used in modern scientific investigations.

项目摘要/摘要 唾液酸酶（也称为神经苷酶）是负责去除唾液酸（SIA）的糖苷酶 来自糖合道的聚糖部分的残基（脱酰化）。根据设计，唾液酸酶能够 调节含SIA的分子的功能和稳定性，并参与 身体和病理途径。上一项和我们最近的研究表明溶酶体NEU1 唾液酸酶可以在LPS刺激后将巨噬细胞的细胞表面迁移到巨噬细胞的细胞表面，在此引起 TLR4受体的脱酰化，导致TLR4激活并随后产生促炎性 细胞因子。 LPS对TLR4激活的失调负责慢性和急性炎症 通常会引起危险疾病（如败血症）的疾病，仍然缺乏特定的药物学 治疗。该应用程序的第一个目的是定量介绍特定于位置的表达式 对于TLR4激活及其随后的信号转导至关重要的Neu1唾液酸酶。 抑制剂是研究唾液酸酶功能和生物学相关机制的有用工具 途径。更重要的是，有效的唾液酸酶抑制剂可以用作药物来调节病理学 由唾液酸酶引起的途径，例如失调的TLR4激活。我们最近的研究表明 当前可用的锅唾液酸酶抑制剂和微生物唾液酸酶抑制剂无法抑制哺乳动物 有效的唾液酸酶。已经报道了几种哺乳动物唾液酸酶抑制剂。但是，当前 唾液酸酶抑制剂设计通常集中于主动位点结合，忽略了亚细胞定位 因此，在活性酶中，它们在体内有效或可能有毒，因为它们会影响其他 细胞内部的唾液酸酶。该应用的第二个目标是开发特定位置的抑制剂 对于neu1唾液酸酶，并定义了neu1 sialidase参与LPS/TLR4信号通路。这 该项目的目标将通过三个特定目的来实现：（1）剖面neu1 sialidase表达式 LPS刺激后巨噬细胞中的细胞表面迁移； （2）发展溶酶体靶向neu1 在LPS/TLR4信号通路中有效调节的唾液酸酶抑制剂； （3）发展细胞 表面靶向Neu1唾液酸酶抑制剂，用于LPS/TLR4信号传导中有效调节脱酰化 路径。这项研究具有创新性，因为它使用了一种克服当前局限性的新方法 在（a）分析唾液酸酶的表达和恢复以及（b）在亚细胞位置抑制唾液酸酶。这 拟议的项目很重要，因为它将（i）发现特定的脱酰基化，这对 LPS/TLR4信号途径和（ii）开发新型的唾液酸酶抑制剂，以有效调节被抑制剂 在LPS/TLR4信号通路中。最后，该提案将增强研究基础设施和 克利夫兰州立大学的教育，允许本科生学习一系列 现代科学研究中使用的实验技术，数据分析和演示技巧。