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: 9540935
• 负责人: SHANJANA AWASTHI
• 金额: $ 36.25万
• 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/9540935
• 关键词: Adult Respiratory Distress Syndrome; Amino Acid Motifs; Amino Acids; Anti-Inflammatory Agents; Anti-inflammatory; Antibiotics; Antiinflammatory Effect; 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; Lung Inflammation; Lung infections; 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; lead candidate; 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.

项目概要 肺部感染和炎症是全世界发病和死亡的主要原因。 感染及其相关的炎症损伤导致急性呼吸窘迫综合征（ARDS）， 没有可用的特异性治疗方法，理想的治疗方法是能够控制病原体负荷的方法。 并使用计算、免疫化学和功能筛选来抑制炎症反应。 方法，一种 Toll 样受体 (TLR4) 相互作用的表面活性剂蛋白 A 衍生肽，称为 SPA4（氨基 SPA4 肽与 TLR4 结合并抑制。 TLR4 诱导的炎症，但维持 TLR4 诱导的病原体识别、摄取和 该项目假设 SPA4 肽将减少病原体负担。 通过与肺部感染和炎症相互作用并改变炎症反应 TLR4复合物的组装具体目标是：（1）定义促吞噬细胞的分子基础。 SPA4肽的抗炎作用，（2）评价SPA4肽在小鼠体内的生物学效应 肺部感染和炎症模型，(3) 分析 SPA4 肽对 TLR4 信号传导的活性和 我们将确定对结合至关重要的 SPA4 肽的氨基酸和基序。 TLR4 和针对感染刺激的活性 我们将研究 TLR4 及其接头分子的组装。 使用结构生物学、计算、分子、生物化学和细胞在人体细胞和模型中 细菌的摄取和清除以及炎症参数将同时进行。 在铜绿假单胞菌和脂多糖的小鼠模型中进行了评估。 我们将使用诱导的肺部感染和炎症来测试 SPA4 肽的生物学相关性。 研究 SPA4 肽的毒性和免疫原性，以及 SPA4 肽重复后的功效 在免疫抑制小鼠中，在二次攻击模型中，以及在 与传统抗生素、表面活性剂或抗炎剂联合使用可减少细菌负担、炎症。 将评估结合、分布、参数、组织病理学、肺功能、症状和存活率。 将评估 SPA4 肽的药代动力学和活性，以调节 TLR4 依赖性 TLR4及其相关野生型和基因小鼠模型肺细胞和组织水平的机制 接头，使用体内成像、免疫组织化学、流式细胞术和共聚焦显微镜。 预计拟议研究和分析的终点决定因素最终将有助于建立 SPA4肽的作用机制该项目采用了开发新型的独特概念。 我们预计这项研究的结果将促进基于肽的免疫治疗的发展。 SPA4肽作为免疫治疗药物，有助于减轻病原体负荷和炎症，改善临床疗效 肺部感染和炎性损伤患者的病情和生存，并控制ARDS。