Role of microRNA in stored RBC induced innate immune suppression

 描述（由申请人提供）：候选人的长期目标是成为一名独立的翻译研究者，并通过了解储存红细胞（RBC）的免疫抑制作用机制来提高输血的安全性。本建议的目的是为候选人提供早期的翻译研究培训 并确定microRNA作为红细胞储存相关先天免疫抑制介质的作用。先天性免疫抑制在重症儿童中很常见，并与不良结局相关。在重症监护室超过48小时的儿童中，高达49%的儿童将接受RBC输血。红细胞输注已被证明是免疫抑制通过未知的机制。较旧储存的RBC通过能够在mRNA水平上干扰促炎细胞因子产生的热稳定的可溶性介质在体外抑制先天性免疫细胞（单核细胞）功能。微小RNA（microRNA，miRNA）是一种小的、热稳定的、非编码的RNA，能够调控多个基因. RBC含有miRNA，并且特定miRNA的相对丰度可以在RBC储存期间随时间变化。该提议的中心假设是储存的RBC通过特异性miRNA的作用抑制单核细胞功能。具体目标一将全面鉴定储存期间RBC miRNA含量的变化，并确定暴露于储存的RBC产品后单核细胞中基因表达的变化。将使用计算方法来确定可能负责观察到的储存RBC对单核细胞的免疫抑制作用的miRNA。Specific Aim 2将为特异性miRNA的免疫抑制作用提供实验证据。具体目标3将证明miRNA靶向策略预防RBC诱导的单核细胞抑制的可行性。本提交资料中描述的工作将为RBC输注相关的先天免疫抑制提供新的机制见解。这些研究的进行和基础科学技术，分子免疫学，microRNA生物学，生物统计学，生物信息学和输血医学的具体指导将支持候选人向独立过渡。

 DESCRIPTION (provided by applicant): The candidate's long term goals are to be an independent translational investigator and to improve the safety of blood transfusion by understanding mechanisms of immunosuppressive effects of stored red blood cells (RBCs). The objectives of this proposal are to provide the candidate with early translational research training and to determine the role of microRNA as mediators of red blood cell storage-related innate immune suppression. Innate immune suppression is common in critically ill children and is associated with adverse outcomes. Up to 49% of children who are in the intensive care unit for greater than 48 hours will receive an RBC transfusion. RBC transfusion has been shown to be immunosuppressive through unknown mechanisms. Older stored RBCs suppress innate immune cell (monocyte) function in vitro via heat-stable, soluble mediators capable of interfering with pro-inflammatory cytokine production at the mRNA level. MicroRNA (miRNA) is small, heat- stable, non-coding RNA capable of regulating multiple genes. RBCs contain miRNA and the relative abundance of specific miRNA may change over time during RBC storage. The central hypothesis of this proposal is that stored RBCs suppress monocyte function via the action of specific miRNA. Specific Aim one will comprehensively identify changes in RBC miRNA content over storage duration and determine changes in gene expression in monocytes after exposure to variably aged RBC products. A computational approach will be used to determine miRNA likely responsible for observed immunosuppressive effects of stored RBCs on monocytes. Specific Aim 2 will provide experimental evidence for immunosuppressive effects of specific miRNA. Specific Aim 3 will demonstrate the feasibility of a miRNA- targeted strategy to prevent RBC-induced monocyte suppression. The work described in this submission will lend novel mechanistic insights into RBC transfusion-related innate immune suppression. The conduct of these studies and specific instruction in basic science techniques, molecular immunology, microRNA biology, biostatistics, bioinformatics, and transfusion medicine will support the candidate's transition to independence.