Role of peptidylarginine deiminase in modulating macrophage functions in sepsis

肽基精氨酸脱亚氨酶在调节脓毒症巨噬细胞功能中的作用

基本信息

批准号：
10387837
负责人：
Jessie W Ho
金额：
$ 7.43万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-01 至 2024-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10387837
关键词：
Ablation Acute Affect Antibiotics Arginine Biological Markers Bone Marrow Calcium Candidate Disease Gene Caring Cause of Death Cell Death Cessation of life Citrulline Complex Complication Dependovirus Disease Employment Enzymes Etiology Family Financial compensation Foundations Functional disorder Genes Genetic Genetic Transcription Guide RNA Histone H3 Histones Hospitals IV Fluid Immune Immune response Infection Inflammation Inflammatory Knock-out Knowledge Lead Life Link Mediating Modeling Morbidity - disease rate Multiple Organ Failure Mus Nuclear Organ Outcome Pathogenesis Pathway interactions Patients Physiological Post-Translational Protein Processing Protein-arginine deiminase Proteomics Pseudomonas aeruginosa Public Health RNA Research Resistance Role Sepsis Serum Signal Transduction Supportive care Tissues United States United States National Institutes of Health Wild Type Mouse Work candidate identification cecal ligation puncture cost effective therapy experimental study genetic profiling improved improved outcome inhibitor insight macrophage mortality mouse model neutrophil novel therapeutic intervention organ injury prevent protein expression restoration sepsis induced acute lung injury septic septic patients targeted treatment tool transcriptome sequencing

项目摘要

Project Summary Sepsis is a life-threatening organ dysfunction caused by a multifaceted dysregulated host response to an infection, including sepsis induced acute lung injury (sepsis-ALI). Sepsis is the most common cause of in- hospital deaths and costs more than $24 billion annually in the US. No specific therapy exists to prevent or treat the cellular dysfunction associated with sepsis and sepsis-ALI. While the complex mechanisms of sepsis have remained poorly defined, an emerging pathogenesis of sepsis and sepsis-ALI involves neutrophil and macrophage cell death causing inflammation and organ damage through the release of citrullinated histone 3 release (CitH3). Petidylarginine deiminase (PAD) 2 and 4 catalyze the formation of CitH3. PAD2 and CitH3 have been linked to sepsis with findings of elevated serum levels of PAD2 and CitH3 in septic patients. Mouse models have further demonstrated that knockout of Pad2 and utilization of pan-PAD inhibitors improved survival and prevented organ dysfunction in several different sepsis models. Throughout the past decade, Dr. Alam and Dr. Li have conducted extensive work on the PAD-CitH3 pathway in sepsis. This proposal is linked to the NIH R01 proposal (1R01HL155116-01A1) “PAD2 and CitH3 in Pathogenesis of Sepsis-induced ALI” by Dr. Alam and Dr. Li with a specific focus on elucidating the mechanisms of PAD and their physiologic impacts and modulation of macrophages in sepsis. In Aim 1, we will conduct experiments using Pad 2-/-, Pad4-/-, and Pad2-/-Pad4-/- mice to investigate the interaction of PAD2 and PAD4 in sepsis and sepsis-ALI outcomes. In Aim 2, we will use adeno-associated virus (AAV)- mediated delivery of Cas9/gRNA to perform targeted knockout and restoration of Pad2 in macrophages. Outcomes of mice with sepsis and their macrophage activity with somatic ablation and targeted RNA knockout will be compared to assess possible compensatory changes in mice with somatic ablation. For Aim 3, we will use RNA sequencing and proteomics to identify genes associated with the compensation of Pad2 ablation. The knowledge gained from this project will have a significant impact on our understanding of the PAD-CitH3 pathway in sepsis. This will contribute significant insight in to the pathogenesis and potential treatment of sepsis, sepsis-ALI, and sepsis induced multi-organ dysfunction.

项目概要脓毒症是一种危及生命的器官功能障碍，由多方面的宿主反应失调引起感染，包括脓毒症引起的急性肺损伤（脓毒症-ALI），脓毒症是感染的最常见原因。在美国，每年造成的医院死亡和费用超过 240 亿美元，目前尚无具体的治疗方法可以预防或预防该疾病。治疗与脓毒症和脓毒症-ALI 相关的细胞功能障碍，而脓毒症的复杂机制。目前尚不清楚，脓毒症和脓毒症-ALI 的新发病机制涉及中性粒细胞和巨噬细胞死亡通过释放瓜氨酸组蛋白 3 引起炎症和器官损伤释放 (CitH3)。哌替啶精氨酸脱亚胺酶 (PAD) 2 和 4 催化 CitH3 和 CitH3 的形成。脓毒症小鼠血清 PAD2 和 CitH3 水平升高与脓毒症有关。模型进一步证明 Pad2 的敲除和泛 PAD 抑制剂的利用得到改善在过去的十年中，博士在几种不同的脓毒症模型中存活并预防了器官功能障碍。 Alam 和 Li 博士对败血症中的 PAD-CitH3 通路进行了广泛的研究。 NIH R01 提案 (1R01HL155116-01A1) “PAD2 和 CitH3 在脓毒症诱发的 ALI 发病机制中的作用”，作者：Dr. Alam 和 Li 博士特别关注阐明 PAD 的机制及其生理影响和败血症中巨噬细胞的调节在目标 1 中，我们将使用 Pad 2-/-、Pad4-/- 和进行实验。 Pad2-/-Pad4-/- 小鼠研究 PAD2 和 PAD4 在脓毒症和脓毒症 ALI 结局中的相互作用。目标 2，我们将使用腺相关病毒 (AAV) 介导的 Cas9/gRNA 递送来执行靶向脓毒症小鼠及其巨噬细胞中 Pad2 的敲除和恢复。将比较体细胞消融和靶向 RNA 敲除的活性，以评估可能的补偿对于目标 3，我们将使用 RNA 测序和蛋白质组学来识别小鼠的体细胞消融变化。从该项目中获得的知识将具有与 Pad2 消融补偿相关的基因。对我们对脓毒症中 PAD-CitH3 通路的理解产生重大影响，这将做出重大贡献。深入了解脓毒症、脓毒症-ALI 和脓毒症引起的多器官损伤的发病机制和潜在治疗方法功能障碍。