High Throughput Screening Assays to Identify Inhibitors of TLR4 Signaling

用于鉴定 TLR4 信号传导抑制剂的高通量筛选试验

• 批准号: 8050426
• 负责人: PETER S TOBIAS
• 金额: $ 18.99万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-27 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8050426
• 关键词: Alzheimer' s Disease; Amyloid; Atherosclerosis; Basic Science; Binding; Biological Assay; Biological Models; CD36 gene; Cell Line; Chinese Hamster Ovary Cell; Chronic; Complement; Deposition; Dimerization; Disease; Endotoxins; Enzymes; Family; Gene Deletion; Goals; Heterodimerization; Homodimerization; Human; Immune; In Vitro; Inflammation; Inflammatory Response; Knock-out; Lactamase; Lead; Ligands; Lipoproteins; Pathologic; Pharmaceutical Preparations; Pharmacotherapy; Plant Roots; Research; Role; Signal Transduction; Sterility; TLR2 gene; TLR4 gene; TLR6 gene; Techniques; Technology; Toll-like receptors; Transfection; Work; counterscreen; high throughput screening; inhibitor/antagonist; monomer; novel; oxidized low density lipoprotein; pathogen; receptor; response; small molecule

DESCRIPTION (provided by applicant): The work proposed here seeks to identify small molecules that could be used to validate a novel hypothesis about the origin of inflammation in atherosclerosis and Alzheimer's disease. The hypothesis is that TLR4 together with TLR6 and CD36 respond to the ligands OxLDL and amyloid-? (A?) to promote the inflammation that is at the root of these diseases. Because of the pleiotropic roles for all three of these molecules, standard techniques of gene deletion cannot be used to validate this hypothesis. We will develop enzyme fragment complementation assays using fragments of ?-lactamase fused to TLR4 and TLR6 to enable high throughput screening assays to identify a pool of small molecules with the potential to inhibit TLR4-TLR6 interactions. Counterscreening assays are described to identify which compounds in the pool are specific inhibitors of the binding between TLR4 and TLR6 that do not also inhibit other pairings of TLR4 and TLR6 that are important in contexts other than atherosclerosis and Alzheimer's. These validated compounds will enable further research to explore the hypothesis. If the hypothesis is validated, it will identify a new paradigm for inflammation in atherosclerosis and Alzheimer's disease and could lead to new forms of drug therapy. PUBLIC HEALTH RELEVANCE: Inflammation in atherosclerosis and Alzheimer's disease is a major cause of the pathological consequences of these diseases. This work will explore new ideas about the origin of the inflammation. It will identify new compounds to inhibit the inflammation in model systems and could lead to new drugs for human therapy.

描述（由申请人提供）：本文提出的工作旨在鉴定可用于验证关于动脉粥样硬化和阿尔茨海默病炎症起源的新假设的小分子。假设 TLR4 与 TLR6 和 CD36 一起对配体 OxLDL 和淀粉样蛋白-？ (A?) 促进炎症，这是这些疾病的根源。由于所有这三种分子的多效性作用，基因删除的标准技术不能用于验证这一假设。我们将使用与 TLR4 和 TLR6 融合的 β-内酰胺酶片段开发酶片段互补测定，以实现高通量筛选测定，从而鉴定出具有抑制 TLR4-TLR6 相互作用潜力的小分子库。描述了反筛选测定法，以确定池中哪些化合物是 TLR4 和 TLR6 之间结合的特异性抑制剂，这些化合物不会抑制 TLR4 和 TLR6 的其他配对，这些配对在动脉粥样硬化和阿尔茨海默病以外的情况下很重要。这些经过验证的化合物将使进一步的研究能够探索这一假设。如果这一假设得到验证，它将确定动脉粥样硬化和阿尔茨海默病炎症的新范例，并可能导致新形式的药物治疗。 公共卫生相关性：动脉粥样硬化和阿尔茨海默氏病的炎症是这些疾病病理后果的主要原因。这项工作将探索关于炎症起源的新想法。它将鉴定出抑制模型系统炎症的新化合物，并可能产生用于人类治疗的新药物。