权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Outer membrane vesicles in macrophage inflammation and pyroptosis

巨噬细胞炎症和焦亡中的外膜囊泡

基本信息

批准号：
10365997
负责人：
Suzhao Li
金额：
$ 7.78万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-05 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10365997
关键词：
Address Bacteria Bacterial Translocation Binding Biological Assay CASP1 gene CRISPR screen Candidate Disease Gene Caspase Cell Death Cells Clustered Regularly Interspaced Short Palindromic Repeats Complex Cytosol Development Disease Drug Delivery Systems Endocytosis Endotoxins Engineering Enzyme Precursors Genes Genetic Screening Glean Goals Gram-Negative Bacteria Human Immune Individual Infection Inflammasome Inflammation Inflammatory Inflammatory Response Innate Immune Response Interleukin-1 beta Knowledge Light Lipopolysaccharides Measures Mediating Mediator of activation protein Membrane Molecular Mus Pathway interactions Pattern Phospholipids Pilot Projects Research Role Source Surface System TLR4 gene Vesicle combat cytokine delivery vehicle endosome membrane extracellular genome-wide insight macrophage pathogen tool vesicular release

项目摘要

PROJECT SUMMARY Innate immune cells such as macrophages detect pathogens by recognizing pathogen-associated molecular patterns (PAMPs) such as lipopolysaccharides (LPS, also known as endotoxin) displayed on the surface of Gram-negative bacteria. Extracellular and endosomal LPS molecules are recognized by Toll-like receptor 4. Cytosolic LPS, on the other hand, is detected by caspase-11 (in mice) and caspase-4/5 (in humans). Interestingly, it was recently found that outer membrane vesicles (OMVs) released by extracellular Gram-negative bacteria can deliver LPS and other bacterial molecules to the cytosol of macrophages. OMVs first enter macrophages through endocytosis, followed by the translocation of bacterial molecules across the endosome membrane into the cytosol to trigger pyroptosis, an inflammatory form of cell death. The molecular basis of OMV-triggered pyroptosis remains poorly understood. In particular, it is unknown how OMVs bind and enter macrophages, and how OMVs crosses the endosome membrane. The goal of this proposed research is to address these key questions by genetically dissecting OMV-induced pyroptosis using an unbiased genome-wide CRISPR genetic screen. In our preliminary studies, we established assays to measure OMV-induced pyroptosis in macrophages. Moreover, we developed tools to genetically dissect complex mammalian pathways using genome-wide CRISPR screens. Here, we will take strategic advantage of these systems to dissect OMV- induced pyroptosis in macrophages using an unbiased genome-wide CRISPR genetic screen. We will then validate the identified genes in pooled and targeted secondary screens. Finally, we will further characterize selected genes identified in the screens in primary human macrophages. These studies will provide a genome-scale view of OMV-induced pyroptosis and will expand our knowledge of inflammatory responses in pathogen-host interactions. Insights gleaned from this pilot study will facilitate the development of safer and more effective strategies to treat pathogen infection and inflammatory diseases. Moreover, we anticipate that these findings will guide the engineering of drug delivery vehicles using OMVs.

项目总结