Regulatory Role of Splicing In Inflammation

项目摘要 炎症的调节是至关重要的，无论是对于治疗的进展， 干预和限制有害的自身免疫并发症。精确的调音 的炎症反应涉及转录，从我们最近未发表的研究， 在许多水平上对数百种mRNA进行细致的调控。自从我们发现NF-κB在 1986年，我们一直在研究这种转录因子系统的各种性质， 我们的注意力主要集中在NF-κB在免疫系统中的作用，特别是在 协调对病原体攻击的炎症反应。近年来我们 研究了在基因表达过程中支持基因表达精确时间的调控事件， 炎症反应。我们使用RNA-seq仅靶向炎症转录物， 炎症基因内含子的定量剪接动力学，发现一些是 拼接速度慢于预期。我们发现它们能显著降低 基因表达作为剪接中的延迟通常伴随着RNA外泌体接合。 我们预测这可能是一种调节机制，并称之为“瓶颈内含子”。在这 建议，我们建议在组织培养中测试瓶颈内含子的生物相关性， 通过制造小鼠（目标1）来观察在修复的背景下炎症的限度是否改变， 内含子。此外，我们建议调查是否有生物背景（刺激， 细胞类型，发育状态），赋予改善的瓶颈剪接（目标2），因此 为这一发现提供了一个监管框架。

PROJECT SUMMARY Regulation of inflammation is of crucial importance, both for the advancement of therapeutic intervention and for the limiting of deleterious autoimmune complications. The precise tuning of the inflammatory response involves transcription and, from our recent unpublished studies, meticulous regulation of hundreds of mRNAs at many levels. Since we discovered NF-κB in 1986, we have been examining a variety of properties of this transcription factor system, focusing most of our attention on the role of NF-κB in the immune system, particularly in orchestrating the inflammatory response to pathogen challenge. In recent years, we have studied the regulatory events underpinning the precise timing of gene expression during the inflammatory response. We used RNA-seq to target only inflammatory transcripts and have quantified splicing kinetics of introns of inflammatory genes, finding that some are orders of magnitude slower to splice than expected. We find that they confer a significant reduction in gene expression as delays in splicing are often concomitant with RNA exosome engagement. We predict this may be a regulatory mechanism, and call them ‘bottleneck introns.’ In this proposal, we propose to test the biological relevance of bottleneck introns in tissue culture and by making mice (Aim 1) to see if limits to inflammation are altered in the context of a repaired intron. In addition, we propose to investigate whether there are biological contexts (stimulus, cell-type, developmental state) that confers improved splicing of a bottleneck (Aim 2), therefore providing a regulatory framework for this finding.