Surfactant protein-A regions as TLR4-immunomodulators

表面活性剂蛋白 A 区域作为 TLR4 免疫调节剂

• 批准号: 9394283
• 负责人: SHANJANA AWASTHI
• 金额: $ 34万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-08 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9394283
• 关键词: Adult Respiratory Distress Syndrome; Amino Acid Motifs; Amino Acids; Anti-Inflammatory Agents; Anti-inflammatory; Antibiotics; Bacteria; Binding; Biochemical; Biological; Biological Testing; Cell model; Cells; Clinical; Clinical Trials; Complex; Computer Analysis; Computer Simulation; Confocal Microscopy; Cytolysis; Development; Docking; Drug Kinetics; Failure; Flow Cytometry; Formulation; Functional disorder; Genetic; Goals; Host Defense; Human; Immune; Immunohistochemistry; Immunomodulators; Immunotherapeutic agent; Infection; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Injury; Lead; Lipopolysaccharides; Lung; Measures; Modeling; Molecular; Morbidity - disease rate; Mus; N-terminal; Organ; Pathology; Patients; Peptide Fragments; Peptides; Phagocytes; Pharmacotherapy; Population; Property; Proteins; Pseudomonas aeruginosa; Public Health; Pulmonary Surfactant-Associated Protein A; Receptor Signaling; Respiratory physiology; Signal Transduction; Stimulus; Symptoms; TLR4 gene; Testing; Therapeutic; Tissues; Toll-like receptors; Toxic effect; Translating; base; cost; cytokine; immune function; immunogenicity; immunoregulation; immunosuppressed; improved; in vivo imaging; mortality; mouse model; mutant; novel; novel therapeutics; pathogen; screening; structural biology; surfactant; uptake

Project Summary Lung infections and inflammation are major causes of morbidity and mortality worldwide. Uncontrolled lung infection and its associated inflammatory injuries lead to acute respiratory distress syndrome (ARDS), for which no specific treatment is available. An ideal therapeutic would be one that can control pathogen burden and suppress the inflammatory response. Using computational, immunochemical, and functional screening approaches, a Toll-like receptor (TLR4)-interacting surfactant protein-A derived peptide called SPA4 (amino acids: GDFRYSDGTPVNYTNWYRGE) has been identified. The SPA4 peptide binds to the TLR4, suppresses the TLR4-induced inflammation, and yet maintains the TLR4-induced pathogen-recognition, uptake and intracellular processing. The project hypothesis is that the SPA4 peptide will reduce the pathogen burden and inflammatory response during lung infection and inflammation by interacting with and altering the assembly of the TLR4 complex. The specific aims are to: (1) define the molecular basis of the pro-phagocytic and anti-inflammatory effects of the SPA4 peptide, (2) evaluate the biological effects of SPA4 peptide in mouse models of lung infection and inflammation, and (3) analyze the activity of SPA4 peptide on TLR4-signaling and immune function. We will determine the amino acids and motif of SPA4 peptide that are critical for binding to TLR4 and activity against infectious stimuli. We will investigate the TLR4-assembly with its adaptor molecules in human cells and models using structural biology, computational, molecular, biochemical, and cellular approaches. The bacterial uptake and clearance and inflammatory parameters will be simultaneously assessed in primary human cells. Mouse models of Pseudomonas aeruginosa- and lipopolysaccharide- induced lung infection and inflammation will be used to test the biological relevance of SPA4 peptide. We will investigate the SPA4 peptide’s toxicity and immunogenicity, and efficacy of SPA4 peptide upon repeat administration, in immunosuppressed mice, in secondary challenge model, and when administered in combination with conventional antibiotic, surfactant, or anti-inflammatory agent. Bacterial burden, inflammatory parameters, tissue pathology, lung function, symptoms and survival will be assessed. The binding, distribution, pharmacokinetics, and activity of SPA4 peptide will be evaluated for modulation of TLR4-dependent mechanisms at cellular and tissue levels in lungs of wild type and genetic mouse models of TLR4 and its adaptors, using in vivo imaging, immunohistochemistry, flow cytometry, and confocal microscopy. It is expected that the end-point determinants of the proposed studies and analyses will eventually help establish the mechanism of action of the SPA4 peptide. This project uses the unique concept of developing a novel peptide-based immunotherapeutic. We anticipate that the results of this study will facilitate the development of SPA4 peptide as an immunotherapeutic to help reduce pathogen burden and inflammation, improve the clinical condition and survival of patients suffering from lung infection and inflammatory injuries, and control ARDS.

项目摘要 肺部感染和炎症是全球发病率和死亡率的主要原因。不受控制的肺 感染及其相关的炎性损伤导致急性呼吸窘迫综合征（ARDS）， 没有具体的治疗方法。一种理想的治疗方法是能够控制病原体负荷 抑制炎症反应使用计算机、免疫化学和功能筛选 Toll样受体（TLR 4）相互作用的表面活性蛋白-A衍生肽SPA 4（氨基 酸：GDFRYSDGTPVNYTNWYRGE）。SPA 4肽与TLR 4结合，抑制TLR 4的表达， TLR 4诱导的炎症，但仍维持TLR 4诱导的病原体识别，摄取和 细胞内加工项目假设是SPA 4肽将减少病原体负荷 以及肺部感染和炎症过程中的炎症反应， TLR 4复合物的组装。具体目的是：（1）明确促吞噬细胞的分子基础 （2）评价SPA 4肽在小鼠体内的生物学效应 肺感染和炎症模型，以及（3）分析SPA 4肽对TLR 4-信号传导的活性， 免疫功能我们将确定SPA 4肽中对于结合至关重要的氨基酸和基序 TLR 4和抗感染刺激的活性。我们将研究TLR 4与其接头分子的组装 在人体细胞和模型中，使用结构生物学、计算、分子、生物化学和细胞生物学， 接近。细菌摄取和清除以及炎性参数将同时被测量。 在原代人类细胞中进行评估。绿脓杆菌和脂多糖的小鼠模型 诱导的肺部感染和炎症将用于测试SPA 4肽的生物相关性。我们将 研究了SPA 4肽的毒性和免疫原性，以及SPA 4肽在重复给药后的功效 在免疫抑制小鼠中，在二次激发模型中，以及在 与常规抗生素、表面活性剂或抗炎剂组合。细菌负荷，炎症 将评估参数、组织病理学、肺功能、症状和存活率。绑定，分配， 将评价SPA 4肽的药代动力学和活性以调节TLR 4依赖性的细胞内信号传导。 在TLR 4的野生型和遗传小鼠模型的肺中的细胞和组织水平的机制及其 衔接子，使用体内成像、免疫组织化学、流式细胞术和共聚焦显微镜。是 预计拟议研究和分析的终点决定因素最终将有助于建立 SPA 4肽的作用机制。这个项目采用了独特的概念， 基于肽的免疫系统。我们预计，这项研究的结果将有助于发展 SPA 4肽作为一种免疫调节剂，有助于减少病原体负担和炎症，改善临床 病情和生存的患者患有肺部感染和炎性损伤，并控制ARDS。