Anti-inflammatory activity of hydrogels designed to capture extracellular inflammasomes

旨在捕获细胞外炎症小体的水凝胶的抗炎活性

• 批准号: 10746957
• 负责人: Eva de Alba Bastarrechea
• 金额: $ 22.22万
• 依托单位: UNIVERSITY OF CALIFORNIA, MERCED
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10746957
• 关键词: Adaptor Signaling Protein; Adopted; Amino Acid Sequence; Anti-Inflammatory Agents; Atherosclerosis; Binding; Biochemical; CASP1 gene; Cardiovascular Diseases; Cell Death; Cell membrane; Cell-Free System; Cells; Cellular biology; Chronic; Chronic Obstructive Pulmonary Disease; DNA; Data; Dependence; Detection; Diabetes Mellitus; Diameter; Disease; Environment; Enzyme-Linked Immunosorbent Assay; Filament; Flow Cytometry; Fluorescence; Fluorescence Microscopy; Fluorescence Spectroscopy; Functional disorder; Funding; Future; Glues; Hydrogels; Immune; Implant; Infection; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Injectable; Injury; Knowledge; Label; Laboratories; Life; Macrophage; Malignant Neoplasms; Measures; Molecular; Monitor; Multiprotein Complexes; Organ; Pathogen detection; Pathogenicity; Penetration; Peptide Hydrolases; Pharmaceutical Preparations; Play; Protein Engineering; Protein Isoforms; Proteins; Public Health; Regulation; Reporting; Research; Resolution; Rheumatoid Arthritis; Role; Rupture; SARS-CoV-2 infection; Signal Transduction; Sodium Chloride; Structure; Technology; Temperature; Testing; Time; Tissues; United States National Institutes of Health; Western Blotting; cellular imaging; cytokine; cytokine release syndrome; design; direct application; experience; extracellular; human disease; macromolecular assembly; meter; nervous system disorder; optic trap; optical traps; particle; pathogen; protein complex; protein protein interaction; recruit; self assembly; sensor; therapeutic target; three dimensional structure; ultra high resolution

PROJECT SUMMARY/ABSTRACT Multiprotein complexes known as inflammasomes form in innate immune cells to trigger inflammation upon detection of pathogens or tissue damage. Abnormal inflammasome activation leads to chronic inflammation, which is the culprit of numerous life-threatening diseases such as cancer, diabetes, cardiovascular disorders, and the cytokine storm in SARS-CoV-2 infection. Inflammasome assembly is controlled by protein-protein interactions as it requires the self-association and oligomerization of multiple copies of three proteins: sensors to detect danger signals, a protease to activate inflammatory factors, and the adaptor protein ASC to connect sensor and protease. Inflammasome formation leads to plasma membrane rupture and concomitant cell death, thus resulting in the release of proinflammatory cytokines and inflammasome particles to the extracellular environment. These extracellular inflammasomes are internalized by nearby cells to perpetuate and amplify the inflammatory response. Removing or sequestering extracellular inflammasomes will likely inhibit or reduce inflammation. Therefore, extracellular inflammasomes are potential therapeutic targets. Our laboratory’s extensive experience on the function and structure of the adaptor ASC, and its interactions with other inflammasome proteins, has led us to create hydrogels designed to form specific protein-protein interactions with inflammasomes; thus, they have the potential to broadly inhibit inflammation by effectively capturing and removing extracellular inflammasomes. This project focuses on identifying the hydrogelation factors leading to optimum biding of inflammasome particles in cell-free systems (Aim 1); and determining the anti-inflammatory efficiency of the hydrogels in the presence of activated innate immune cells (Aim 2). Our experimental plan will combine cell biology and biochemical approaches, including live/dead cell imaging, flow cytometry, immunoblotting, enzyme-linked immunosorbent assays and fluorescence spectroscopy. Overall, we expect to develop a hydrogel technology of broad applicability to reduce inflammation in the absence of drug loading by targeting the inflammasome, which is implicated in many inflammatory diseases.

项目摘要/摘要 在先天免疫细胞中形成的称为炎性症的多蛋白络合物，以触发炎症 检测病原体或组织损伤。异常炎性体激活导致慢性炎症， 这是许多威胁生命的疾病的罪魁祸首，例如癌症，糖尿病，心血管疾病， 以及SARS-COV-2感染中的细胞因子风暴。炎性组组件由蛋白质蛋白质控制 相互作用需要三种蛋白质的多个副本的自我关联和寡聚化：传感器 要检测危险信号，一种激活炎症因子的蛋白质以及衔接蛋白ASC连接 传感器和蛋白酶。炎性体的形成导致质膜破裂和伴随的细胞死亡， 因此，导致促炎细胞因子和炎症体颗粒释放到细胞外 环境。这些细胞外炎症体被附近的细胞内部化，以使并扩大 炎症反应。去除或隔离细胞外炎症可能会抑制或减少 炎。因此，细胞外炎症是潜在的治疗靶标。我们的实验室 关于适配器ASC的功能和结构的丰富经验及其与其他的相互作用 炎性蛋白使我们产生了旨在与特定蛋白质 - 蛋白质相互作用的水凝胶 炎症；因此，他们有可能通过有效捕获和 去除细胞外炎症。该项目着重于确定导致 无细胞系统中炎性体颗粒的最佳体屈（AIM 1）；并确定抗炎药 在活化的先天免疫细胞存在下水凝胶的效率（AIM 2）。我们的实验计划将 结合细胞生物学和生化方法，包括活细胞成像，流式细胞术， 免疫印迹，酶联免疫吸附测定和荧光光谱。总体而言，我们希望 在没有药物负荷的情况下，开发具有广泛适用性的水凝胶技术，以减少炎症 针对炎症体，该炎症体在许多炎症性疾病中实施。