Long Non-Coding RNA, TLR Tolerance and Sepsis

长非编码 RNA、TLR 耐受性和脓毒症

基本信息

批准号：
9297691
负责人：
ANDREI E MEDVEDEV
金额：
$ 19.94万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-01-01 至 2018-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9297691
关键词：
Address Affect Agonist American Anti-Inflammatory Agents Anti-inflammatory Bacteria Cells Cessation of life Clinical Trials Data Dendritic Cells Development Disease Disseminated Intravascular Coagulation Endotoxins Epithelial Cells Exhibits Failure Future Genetic Polymorphism Genetic Transcription Grant Health Human IRAK1 gene IRAK3 gene IRAK4 gene Immune Immunocompromised Host Immunoprecipitation Immunosuppression Impairment Infection Inflammation Mediators Inflammatory Intensive Care Units Interleukin-1 alpha Mass Spectrum Analysis Mediating Mediator of activation protein Mission Molecular Molecular Target Mus Myeloid Cells National Institute of Allergy and Infectious Disease Natural Immunity Organ Pathway interactions Patients Pattern Phase Phenotype Post-Transcriptional Regulation Post-Translational Protein Processing Publishing Receptor Signaling Regulation Role Secondary to Sepsis Shapes Signal Pathway Signal Transduction TLR4 gene Testing Toll-like receptors Transcript Translational Research Untranslated RNA antimicrobial cytokine design differential expression drug testing effective therapy experimental study improved knock-down macrophage microbial monocyte mortality next generation sequencing novel strategies pathogen prevent programs response secondary infection septic

项目摘要

Sepsis remains one of the major health threats in the U. S., affecting ~750,000 Americans per year, with mortality rate up to 50%. Many septic patients survive the initial “cytokine storm” but develop profound immunosuppression, become immunocompromised and succumb to secondary infections. Monocytes from such immunocompromised septic patients have “re- programmed” Toll-like receptor (TLR) 4 signaling, showing suppressed pro-inflammatory cytokines but unchanged or increased expression of anti-inflammatory and anti-microbial mediators, reminiscent of endotoxin tolerance. Long non-coding RNAs (lncRNAs) have recently emerged as new regulators of TLR signaling pathways, but how changes in lncRNAs “shape” re- programming of responses of myeloid cells during TLR tolerance and sepsis is unknown. Our preliminary and published data has identified lncRNAs in human THP1 monocytes and mouse macrophages with significant changes in expression upon TLR4 challenge and endotoxin tolerization. We showed that knock-down of PCGEM1 lncRNA in THP-1 cells and deficiencies of natural antisense transcript-IL-1α, linc-Cox2 or Eps lncRNAs in mouse macrophages significantly affect LPS signaling. We also found that Eps-/- Ms are compromised in the induction of endotoxin tolerance. These data supports our hypothesis that altered expression and functions of lncRNAs in myeloid cells promote TLR tolerance and sepsis-associated immunosuppression by reprogramming TLR responses at the level transcriptional and post-transcriptional regulation of inflammatory mediators. To test this hypothesis, we have designed the following Specific Aims: 1. Identify the impact of TLR4 tolerance and sepsis on expression of lncRNAs in myeloid cells; and 2. Determine the functional significance for lncRNAs in TLR4 signaling and tolerance. Having completed this project, we will determine the role for lncRNAs in “reprogramming” of TLR signaling in myeloid cells during development of TLR tolerance and sepsis. This project is expected to improve our understanding of the mechanisms by which lncRNAs modulate antimicrobial responses of myeloid cells, and define molecular targets of lncRNAs for future translational research in human patients with sepsis. These advances would be of key importance for improving treatment of sepsis in the U.S.

脓毒症仍然是美国主要的健康威胁之一，影响约 75 万美国人每年，死亡率高达 50%，许多脓毒症患者在最初的“细胞因子风暴”中幸存下来。但会产生严重的免疫抑制，变得免疫功能低下并屈服于来自此类免疫功能低下的脓毒症患者的单核细胞会“重新感染”。程序化的“Toll 样受体 (TLR) 4 信号传导，显示出抑制的促炎作用细胞因子但抗炎和抗微生物表达不变或增加介质，让人想起内毒素耐受性，最近出现了长非编码RNA（lncRNA）。成为 TLR 信号通路的新调节因子，但 lncRNA 的变化如何“塑造”重新 TLR 耐受和败血症期间骨髓细胞反应的编程尚不清楚。初步和已发表的数据已鉴定出人类 THP1 单核细胞和小鼠中的 lncRNA 巨噬细胞在 TLR4 攻击和内毒素耐受后表达发生显着变化。我们发现 THP-1 细胞中 PCGEM1 lncRNA 的敲低和天然反义的缺陷小鼠巨噬细胞中的转录-IL-1α、linc-Cox2 或 Eps lncRNA 显着影响 LPS 信号传导。我们还发现 Eps-/- Ms 在内毒素耐受性的诱导中受到损害。支持我们的假设，即骨髓细胞中 lncRNA 的表达和功能促进通过重新编程 TLR 反应来实现 TLR 耐受和脓毒症相关免疫抑制炎症介质的转录和转录后调节水平以测试这一点。根据假设，我们设计了以下具体目标： 1. 确定 TLR4 耐受性的影响和脓毒症对骨髓细胞中 lncRNA 表达的影响；以及 2. 确定其功能意义；完成该项目后，我们将确定 lncRNA 在 TLR4 信号传导和耐受中的作用。在 TLR 耐受性发展过程中，lncRNA 参与骨髓细胞 TLR 信号传导的“重编程” 该项目预计将提高我们对脓毒症机制的理解。 lncRNA 调节骨髓细胞的抗菌反应，并定义 lncRNA 的分子靶点对于未来人类脓毒症患者的转化研究来说，这些进展将是关键。改善美国脓毒症治疗的重要性