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（也是平静的神经氨酸酶）是负责去除唾液酸（SIA）的糖苷果。 糖合道的聚糖部分的残留物（通过凝固酶）能够使用。 模块化含SIA的分子的功能和稳定性，并参与 Physiololocal和Patholocal途径。 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 TLR4受体的脱酰化，导致TLR4激活和随后的Pro-炎症 LPS的细胞因子失调TLR4激活 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 处理。 Neu1唾液酸酶对TLR4激活至关重要，是随后的信号交易。 抑制剂是研究唾液酸酶功能和生物学相关机制的我们的工具 更重要的是，有效的唾液酸酶抑制剂可以用作药物 由唾液酸酶引起的途径，例如我们最近的研究指标，例如非正规的TLR4激活。 目前可用的锅唾液酸酶抑制剂和微生物唾液酸酶吸入器无法抑制mamalianan 唾液酸酶有效。 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 开发特定位置抑制剂的第二个目标 对于neu1唾液酸酶，并定义了neu1 sialidase在LPS/TLR4信号通路中的涉及 该项目的目标将由三个特定目的来实现：（1）剖面neu1 sialidase表达式 LPS刺激时巨噬细胞的细胞表面搬迁； 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，以有效调节LPS/TLR4信号传导 这项研究是创新的，因为它使用了一种新颖的方法。 在（a）分析唾液酸酶的表达和迁移以及（b）在亚细胞位置抑制唾液酸酶 支撑项目是重要的，因为它将（i）发现对特定的脱酰化的依赖性。 LPS/TLR4信号途径和（ii）开发新型的唾液酸酶抑制剂，以有效调节脱酰化 在LPS/TLR4信号通路中，该提案提高了研究的基础设施 克利夫兰州立大学的教育，允许地下学生学习广泛的范围 现代科学研究中使用的实验技术，数据分析和演示技巧。