Long Non-Coding RNA, TLR Tolerance and Sepsis

长非编码 RNA、TLR 耐受性和脓毒症

• 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
• 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

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。美国，影响了约75万美国人 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，免疫功能低下， 继发感染。来自这种免疫功能低下的脓毒症患者的单核细胞具有“再- 程序性Toll样受体（TLR）4信号传导，显示抑制的促炎性细胞因子， 但抗炎和抗微生物的表达不变或增加 介质，让人联想到内毒素耐受。长链非编码RNA（lncRNA）最近被 作为TLR信号通路的新调节因子出现，但是lncRNA“形状”的变化如何重新调节TLR信号通路？ TLR耐受和脓毒症期间骨髓细胞应答的编程是未知的。我们 初步和公开的数据已经在人THP 1单核细胞和小鼠单核细胞中鉴定了lncRNA， 在TLR 4攻击和内毒素耐受后，巨噬细胞的表达发生显著变化。 我们发现THP-1细胞中PCGEM 1 lncRNA的敲低和天然反义寡核苷酸的缺乏， 小鼠巨噬细胞中的转录本-IL-1α、linc-Cox 2或Eps lncRNA显著影响LPS信号传导。 我们还发现，Eps-/- M β在诱导内毒素耐受中受到损害。这些数据 支持了我们的假设，即髓系细胞中lncRNA的表达和功能改变促进了 TLR耐受和脓毒症相关的免疫抑制，通过重新编程TLR反应， 水平转录和转录后调节炎症介质。为了验证这一 假设，我们设计了以下具体目标：1。确定TLR 4耐受性的影响 和脓毒症对骨髓细胞中lncRNA表达的影响;和2.确定以下方面的功能意义 TLR 4信号转导和耐受中的lncRNA。完成这个项目后，我们将确定 lncRNA在TLR耐受性形成过程中髓系细胞TLR信号转导的“重编程”中的作用 和败血症。该项目预计将提高我们对机制的理解， lncRNA调节骨髓细胞的抗菌反应，并确定lncRNA的分子靶点 用于未来在脓毒症患者中的转化研究。这些进展将是关键 在美国改善脓毒症治疗的重要性