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：
10393056
负责人：
GEORGE R DUBYAK
金额：
$ 19.28万
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 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10393056
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 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包括与感染相关的毛孔形成蛋白家族炎症和癌症。 RNASEQ分析表明，唯一由GSDM家庭成员表示中性粒细胞是GSDMD和GSDME/DFNA5，初步数据表明，中性粒细胞可以激活caspase- 3，将pro-GSDME切割到孔形成N-GSDME以驱动质膜孔的形成导致促炎性凋亡细胞死亡。这与我们自然传播的结果相反 2020年论文表明，caspase-1和中性粒细胞弹性酶在中性粒细胞中激活的GSDMD发生在没有凋亡细胞死亡。 GSDMD和GSDME都介导了生物活性IL-的中性粒细胞产生 1β，我们报道的是宿主免疫激素中的一个关键细胞因子，需要调节肺炎链球菌和铜绿假单胞菌细菌的生长和角膜无情。我们建议表征GSDME调节和响应肺炎链球菌中嗜中性粒细胞的功能体外和铜绿假单胞菌，并定义了由这些致病性引起的角膜炎中GSDME的体内作用细菌。 AIM 1将表征GSDME调节和在中性粒细胞中的功能，并定义其在响应S. 肺炎或铜绿假单胞菌。 AIM 2将表征受感染角膜中的GSDME处理，并评估 GSDME在肺炎链球菌或铜绿假单胞菌中使用GSDME - / - 和GSDME - / - / - /GSDMD - / - 小鼠中性粒细胞中的作用生存力，细菌杀死和角膜病的严重程度，并将使用自适应转移模型检查中性粒细胞GSDME的作用。拟议研究的结果将洞悉这些孔在中性粒细胞对这些眼病原体的反应中形成蛋白质，并将确定潜力对细菌角膜炎的免疫干预的靶标。