The role of Gasdermin E (DFNA5) in bacterial keratitis

Gasdermin E (DFNA5) 在细菌性角膜炎中的作用

• 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
• 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

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， 炎症和癌症。RNAseq分析显示，仅有的GSDM家族成员表达为： 中性粒细胞是Gsdmd和Gsdme/DFNA 5，初步数据显示中性粒细胞可以激活半胱天冬酶， 3，其将pro-GSDME切割成孔的N-GSDME以驱动质膜孔的形成 导致促炎性细胞死亡。这与我们的《自然通讯》的结果形成了鲜明对比 2020年的论文显示，中性粒细胞中的caspase-1和中性粒细胞弹性蛋白酶激活的GSDMD发生在 不存在细胞死亡。GSDMD和GSDME均介导中性粒细胞产生生物活性IL-1， 1β，我们报道它是宿主免疫应答中的关键细胞因子， 调节肺炎链球菌和铜绿假单胞菌的细菌生长和角膜混浊。 我们建议描述中性粒细胞对S.肺炎 和铜绿假单胞菌在体外的作用，并确定GSDME在这些致病性角膜炎引起的体内作用 细菌目标1将描述GSDME在中性粒细胞中的调节和功能，并定义其在 协调中性粒细胞热凋亡细胞死亡和中性粒细胞胞外陷阱的形成，以响应S. 肺炎假单胞菌或铜绿假单胞菌。目的2将表征感染角膜中的GSDME处理，并评估 GSDME在S.使用GSDME-/-和GSDME-/-/GSDMD-/-小鼠在嗜中性粒细胞中检测肺炎假单胞菌或铜绿假单胞菌 存活力、细菌杀灭和角膜疾病的严重程度，并将使用过继转移模型， 研究中性粒细胞GSDME的作用。拟议研究的结果将使人们深入了解 这些孔形成蛋白在中性粒细胞对这些眼部病原体的反应，并将确定潜在的 细菌性角膜炎免疫干预的靶点。