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: 10225428
• 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 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10225428
• 关键词: Adult Respiratory Distress Syndrome; Amino Acid Motifs; Amino Acids; Anti-Inflammatory Agents; Antibiotics; Antiinflammatory Effect; Bacteria; Binding; Biochemical; Biological; Biological Testing; Cell model; Cells; Clinical; Clinical Trials; Complex; Computer Analysis; 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; Signal Transduction; Stimulus; Structure; Symptoms; TLR4 gene; Testing; Therapeutic; Tissues; Toll-like receptors; Toxic effect; Translating; base; cost; cytokine; immune function; immunogenicity; immunoregulation; immunosuppressed; improved; in silico; in vivo imaging; lead candidate; mortality; mouse model; mutant; novel; novel therapeutics; pathogen; pulmonary function; 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世界发病率和死亡率的主要原因。失控肺 感染及其相关的炎症性损伤导致急性呼吸窘迫综合征(ARDS)， 目前还没有特效的治疗方法。理想的疗法应该是能够控制病原体负担的疗法。 抑制炎症反应。使用计算、免疫化学和功能筛选 方法：一种Toll样受体(TLR4)与表面活性蛋白相互作用的衍生多肽SPA4(氨基 酸：GDFRYSDGTPVNYTNWYRGE)已被鉴定。SPA4多肽与TLR4结合，抑制 TLR4诱导的炎症，但仍维持TLR4诱导的病原体识别、摄取和 细胞内处理。项目假设是SPA4多肽将减少病原体负担 以及肺部感染和炎症过程中的炎症反应 TLR4复合体的组装。具体目的是：(1)确定前吞噬细胞的分子基础 以及SPA4多肽的抗炎作用；(2)评价SPA4多肽在小鼠体内的生物学作用 肺感染和炎症模型；(3)分析SPA4肽对TLR4信号转导和TLR4信号转导的影响。 免疫功能。我们将确定与SPA4多肽结合的关键氨基酸和基序 TLR4和抗感染刺激活性。我们将研究TLR4-组装及其接头分子 在人类细胞和模型中使用结构生物学、计算、分子、生化和细胞 接近了。细菌摄取和清除以及炎症参数将同时进行 在原代人类细胞中进行评估。铜绿假单胞菌和脂多糖的小鼠模型 用诱导的肺部感染和炎症来检验SPA4多肽的生物学相关性。我们会 研究SPA4多肽的毒性、免疫原性及对重复序列的作用 给药，在免疫抑制的小鼠中，在二次激发模型中，当在 与常规抗生素、表面活性物质或消炎剂联合使用。细菌负担，炎症性 将评估参数、组织病理学、肺功能、症状和存活率。绑定、分发、 SPA4多肽的药代动力学和活性将被评估以调节TLR4依赖 TLR4和TLR4基因小鼠模型肺组织和细胞水平的机制 使用活体成像、免疫组织化学、流式细胞术和共聚焦显微镜。它是 预期拟议研究和分析的终点决定因素最终将有助于确定 SPA4多肽的作用机制。这个项目使用了开发一部小说的独特概念 以多肽为基础的免疫治疗。我们预计，这项研究的结果将有助于 SPA4多肽作为一种免疫治疗剂，有助于减轻病原体负担和炎症，改善临床 肺部感染和炎症性损伤患者的病情和生存情况，并控制ARDS。

项目成果

Tight Junctions, the Epithelial Barrier, and Toll-like Receptor-4 During Lung Injury.

• DOI: 10.1007/s10753-022-01708-y
• 发表时间: 2022-12
• 期刊: Inflammation
• 影响因子: 5.1

Mechanism of Anti-Inflammatory Activity of TLR4-Interacting SPA4 Peptide.

• DOI: 10.4049/immunohorizons.2100067
• 发表时间: 2021-08-24
• 期刊: ImmunoHorizons
• 作者: Awasthi S;Kumar G;Ramani V;Awasthi V;Rodgers KK;Xie J;Beierle J;Kyere-Davies G;Singh B;Rahman N;Chowdhury AA;Chataut N
• 通讯作者: Chataut N

Stability and structure-activity relationship of the SPA4 peptide under ambient and stressed conditions of lung injury.

• DOI: 10.1039/d3ra02918b
• 发表时间: 2023-06-15
• 期刊: RSC advances
• 影响因子: 3.9