RNA-binding proteins and control of mRNA metabolism in the regulation of adaptive immunity in humans

RNA结合蛋白和mRNA代谢在人类适应性免疫调节中的控制

• 批准号: 432656284
• 负责人: Professor Dr. Vigo Heissmeyer
• 金额: --
• 依托单位: Forschungsbereich Immunologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2020
• 资助国家: 德国
• 起止时间: 2019-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/432656284?language=en
• 关键词: RNA; binding; proteins; control; mRNA

Post-transcriptional gene regulation can determine mRNA splicing, stability, translation and sub-cellular localization, thereby potentially impacting all cellular functions. In the immune system, the importance of post-transcriptional control in regulating the differentiation and functions of T lymphocytes is becoming increasingly clear, although many questions remain to be addressed. RNA-binding proteins (RBPs) have been variably involved in immune modulation by regulating target mRNA stability and translation. Understanding mechanisms of post-transcriptional regulation of gene expression is particularly important for immune cells, in which the expression of effector molecules such as cytokines must increase rapidly in response to danger signals, but it also must be quickly turned off to avoid excessive inflammation and tissue damage. Indeed, many mRNAs encoding for cytokines and other immune-relevant genes are unstable and/ or kept in a translationally silenced state until needed. However, there are still major gaps in our knowledge regarding the mechanisms of action of RBPs and their functional role in immune cells, especially in humans. Most importantly, the level of cross-interaction, cooperation or redundancy of the different RBPs in regulating the metabolism of specific mRNAs remains almost completely unknown. With this project we will therefore fill these critical knowledge gaps by investigating the following Specific Aims:- Aim 1: To determine the importance of the different RBPs in the differentiation and function of human T lymphocytes. This aim will tackle two major questions: i) what is the impact of mRNA regulation mediated by RBPs specifically in the human system, and ii) to what extent the different family members of a given RBP family have unique and/ or redundant functions. These questions will be addressed primarily by taking advantage of recently optimized systems to perform CRISPR-Cas9-mediated gene deletion in primary human T lymphocytes and by assessing the functional effects of such deletions.- Aim 2: In depth mechanistic analysis of selected RBPs in human T lymphocytes. In this aim we will use a combination of ‘omics’ and targeted approaches to establish the binding profile of selected RPBs in human T cells and their effect on mRNA stability and translation.Overall, we will integrate a comprehensive description of the functional role of selected RBPs in modulating aspects of T lymphocyte biology with a detailed mechanistic analysis of the role of these RBPs in regulating mRNA metabolism during cell activation, differentiation and proliferation. By combining our expertise in the regulation of immune responses by RBPs and in human T lymphocyte biology we aim at unravelling general paradigms linking RBP expression to the regulation of complex cellular responses in human T lymphocytes.

转录后基因调控可以决定mRNA的剪接、稳定性、翻译和亚细胞定位，从而潜在地影响所有细胞功能。在免疫系统中，转录后调控在调节T淋巴细胞分化和功能中的重要性正变得越来越清楚，尽管仍有许多问题有待解决。RNA结合蛋白（RBP）通过调节靶mRNA的稳定性和翻译而参与免疫调节。了解基因表达的转录后调控机制对免疫细胞特别重要，其中效应分子（如细胞因子）的表达必须响应危险信号迅速增加，但也必须迅速关闭以避免过度炎症和组织损伤。事实上，许多编码细胞因子和其他免疫相关基因的mRNA是不稳定的和/或保持在沉默状态直到需要。然而，我们对RBPs的作用机制及其在免疫细胞中的功能作用，特别是在人类中的作用的认识仍然存在重大差距。最重要的是，在调节特定mRNA的代谢中，不同RBP的交叉相互作用、合作或冗余的水平仍然几乎完全未知。因此，通过本项目，我们将通过调查以下具体目标来填补这些关键的知识空白：-目标1：确定不同RBP在人类T淋巴细胞分化和功能中的重要性。这一目标将解决两个主要问题：i）RBP特异性介导的mRNA调控在人类系统中的影响是什么，以及ii）给定RBP家族的不同家族成员在多大程度上具有独特和/或冗余的功能。这些问题将主要通过利用最近优化的系统在原代人类T淋巴细胞中进行CRISPR-Cas9介导的基因缺失以及评估此类缺失的功能效应来解决。目的2：深入分析人T淋巴细胞中选择性RBP的机制。在这个目标中，我们将使用“组学”和靶向方法的组合来建立所选RPB在人类T细胞中的结合谱及其对mRNA稳定性和翻译的影响。总体而言，我们将综合全面描述所选RBP在调节T淋巴细胞生物学方面的功能作用，并详细分析这些RBP在细胞活化期间调节mRNA代谢的作用，分化和增殖。通过结合我们在RBP免疫反应调节和人类T淋巴细胞生物学方面的专业知识，我们的目标是解开RBP表达与人类T淋巴细胞复杂细胞反应调节之间的一般范式。