The role of Gasdermin E (DFNA5) in bacterial keratitis

Gasdermin E (DFNA5) 在细菌性角膜炎中的作用

基本信息

批准号：
10196230
负责人：
GEORGE R DUBYAK
金额：
$ 25.28万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
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：
2021-05-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10196230
关键词：
Adoptive Transfer Autophagocytosis Autophagosome Azurophilic Granule Bacteria Bacterial Infections Biochemical Budgets C57BL/6 Mouse CASP1 gene CASP3 gene Caspase Cell Death Cell Line Cell Membrane Permeability Cell membrane Cells Cessation of life Cleaved cell Clustered Regularly Interspaced Short Palindromic Repeats Communication Cornea Corneal Diseases Corneal Opacity Corneal Stroma Cytoplasmic Granules Cytosol Data Family Family member Funding Genetic Transcription Growth Histones Human Immune Immune Targeting Immune response In Vitro Infection Inflammasome Inflammation Inflammatory Interleukin-1 beta Keratitis Knock-out Leukocyte Elastase Malignant Neoplasms Matrix Metalloproteinases Mediating Mediator of activation protein Membrane Modeling Molecular Mus Nature Neutrophil Activation Organelles Paper Pathogenesis Pathway interactions Peptide Hydrolases Play Principal Investigator Production Protease Inhibitor Protein Family Proteins Pseudomonas aeruginosa Pseudomonas aeruginosa infection Regulation Reporting Resolution Role Serine Serine Protease Serpins Severities Signal Transduction Source Stimulus Streptococcus pneumoniae Tissues VDAC1 gene base corneal scar cytokine extracellular immunological intervention insight member microbial microbicide neutrophil novel pathogen pathogenic bacteria preservation prevent response trafficking transcriptome sequencing

项目摘要

PROJECT ABSTRACT Gasdermin (GSDM) A-E comprise a family of pore-forming proteins that are associated with infection, inflammation and cancer. RNAseq analysis revealed that the only GSDM family members expressed by neutrophils are Gsdmd and Gsdme/DFNA5, and preliminary data show that neutrophils can activate caspase- 3, which cleaves pro- GSDME to the pore-forming N-GSDME to drive formation of plasma membrane pores leading to pro-inflammatory pyroptotic cell death. This is in contrast to results from our Nature Communications 2020 paper showing that that GSDMD activated by caspase-1 and neutrophil elastase in neutrophils occurs in the absence of pyroptotic cell death. Both GSDMD and GSDME mediate neutrophil production of bioactive IL- 1β, which we reported is a pivotal cytokine in the host immune response that is required to regulate Streptococcus pneumoniae and Pseudomonas aeruginosa bacterial growth and corneal opacification. We propose to characterize GSDME regulation and function in neutrophils in response to S. pneumoniae and P. aeruginosa in vitro, and to define the vivo role(s) of GSDME in keratitis caused by these pathogenic bacteria. Aim 1 will characterize GSDME regulation and function in neutrophils and define its role in coordinating neutrophil pyroptotic cell death and formation of neutrophil extracellular traps in response to S. pneumoniae or P. aeruginosa. Aim 2 will characterize GSDME processing in infected corneas, and assess the role of GSDME in S. pneumoniae or P. aeruginosa using GSDME-/- and GSDME-/-/GSDMD-/- mice in neutrophil viability, bacterial killing and the severity of corneal disease, and will use an adoptive transfer model to examine the role of neutrophil GSDME. Results of the proposed studies will provide insight into the role of these pore forming proteins in neutrophil responses to these ocular pathogens, and will identify potential targets for immune intervention in bacterial keratitis.

项目摘要 Gasdermin (GSDM) A-E 包含与感染相关的成孔蛋白家族， RNAseq 分析揭示了 GSDM 家族成员唯一表达的。中性粒细胞为Gsdmd和Gsdme/DFNA5，初步数据表明中性粒细胞可以激活caspase- 3，将 pro-GSDME 裂解为成孔 N-GSDME，以驱动质膜孔的形成导致促炎性焦亡细胞死亡，这与我们的《自然通讯》的结果相反。 2020年论文显示，中性粒细胞中由caspase-1和中性粒细胞弹性蛋白酶激活的GSDMD发生在 GSDMD 和 GSDME 均介导中性粒细胞产生生物活性 IL-。 1β，我们报道它是宿主免疫反应中的关键细胞因子，是调节肺炎链球菌和铜绿假单胞菌细菌生长和角膜混浊。我们建议描述中性粒细胞响应肺炎链球菌的 GSDME 调节和功能特征和铜绿假单胞菌体外，并确定 GSDME 在这些致病菌引起的角膜炎中的体内作用目标 1 将描述 GSDME 在中性粒细胞中的调节和功能，并定义其在细菌中的作用。协调中性粒细胞焦亡细胞死亡和中性粒细胞胞外陷阱的形成以响应金黄色葡萄球菌。目标 2 将表征感染角膜中的 GSDME 处理，并评估使用 GSDME-/- 和 GSDME-/-/GSDMD-/- 小鼠中性粒细胞研究 GSDME 在肺炎链球菌或铜绿假单胞菌中的作用活力、细菌杀灭和角膜疾病的严重程度，并将使用过继转移模型来检查中性粒细胞 GSDME 的作用。拟议研究的结果将提供对中性粒细胞 GSDME 作用的深入了解。这些孔形成蛋白参与中性粒细胞对这些眼部病原体的反应，并将识别潜在的细菌性角膜炎免疫干预的目标。