M.tuberculosis-induced Alternative Processing of IL12RB1 mRNA

结核分枝杆菌诱导的 IL12RB1 mRNA 选择性加工

• 批准号: 8606809
• 负责人: MARK T MCNALLY
• 金额: $ 19.13万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8606809
• 关键词: Affect; Alternative Splicing; Americas; Antigens; Basic Science; Binding; Binding Sites; Biological; Biological Assay; Biology; Cell Surface Receptors; Cell physiology; Cells; Cellular biology; Complex; Cryptococcus neoformans infection; Cytokine Signaling; Cytoplasmic Tail; Disease; Elements; Event; Exposure to; Family; Figs - dietary; Gene Expression; Genetic Polymorphism; Goals; Health; Human; IL12RB1 gene; Immune response; Immunity; Indium; Inflammatory; Interferons; Interleukin-12; Knockout Mice; Knowledge; Lead; Leukocytes; Ligands; Lymphocyte; Measures; Mediating; Messenger RNA; Mission; Molecular; Mouse Strains; Mus; Mycobacterium tuberculosis; Outcome; Pathway interactions; Pattern; Play; Polyadenylation; Population; Prevention; Principal Investigator; Process; Production; Protein Isoforms; Proteins; Public Health; RNA; RNA Processing; Regulation; Relative (related person); Research; Role; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Stimulus; System; T-Lymphocyte; Techniques; Testing; Therapeutic; Therapeutic Intervention; Transmembrane Domain; Tuberculosis; United States; United States National Institutes of Health; Work; base; cytokine; extracellular; human disease; immunogenic; improved; in vitro activity; in vivo; infectious disease treatment; innovation; leukocyte activation; mRNA Precursor; meetings; novel; pathogen; public health relevance; receptor; response; therapeutic target; tuberculosis immunity

DESCRIPTION (provided by applicant): IL12R¿1?TM (Isoform 2) is an alternative IL12 receptor isoform that is expressed by mouse leukocytes following exposure to M. tuberculosis (Mtb). Isoform 2 lacks the transmembrane domain of IL12R¿1 (Isoform 1), has a localization pattern that is distinct from that of Isoform 1, and in contrast to the initial predictions, enhancs IL12-dependent activities in vitro. While it is clear than the activation of human leukocytes with Mtb also stimulates Isoform 2 production via alternative splicing / polyadenylation, there is a gap in our understanding of the factors required for Isoform 2 alternative RNA processing and the significance of this pathway for controlling Mtb infection in vivo. This knowledge gap is important because until filled, avenues for therapeutic manipulation of IL12 activity will remain out of reach. Our central hypothesis is that immunogenic signaling induces alternative splicing/polyA via cis elements in the mRNA to generate Isoform 2, a protein that regulates TB control in vivo by promoting IL12-dependent TH1 differentiation. Our long-term goal is to understand how Mtb stimulates signaling to induce IL12RB1 alternative mRNA processing, and to manipulate Isoform 2 production to influence T cell biology and Mtb control. The objectives in this application are to identify the cis-elements in IL12RB1 pre-mRNA that are the targets of signaling and mediate Isoform 2 production and to test, using human and mouse systems, the significance of Isoform 2 to TB control. The rationale for the proposal is that identification of signal-responsive cis elements in IL12RB1 pre-mRNA that mediate alternative RNA processing, as well as the function of Isoform 2 in the context of TB, will lead to therapeutic targets to modiy pathway activity. The hypothesis will be tested through two specific aims: 1) Identify the cis-elements and trans-factors that mediate Isoform 2 RNA alternative processing and 2) determine the extent to which IL12RB1 Isoform 2 influences Mtb immunity. Aim 1 will utilize IL12R¿1 minigenes that will be nucleofected into T cells to identify cis-elements within the IL12RB1 mRNA that mediate pathogen-induced RNA alternative processing. Roles for candidate processing factors (based on potential binding sites near the regulated polyA site) will be assessed for Isoform 2 alternative processing. Aim 2 will use an IL12-dependent bioassay to test the affect of Isoform 2 silencing in human lymphocytes, as well use a newly generated mouse strain to determine the outcome of experimental TB in the absence of Isoform 2. The work is innovative because it focuses on a completely different level of IL12R¿1 activity regulation - signal-mediated post-transcriptional alternative RNA splicing / polyadenylation - in humans. This work is significant because it is the initial effort towards understanding the earlies events in Isoform 2 production in human cells, which should reveal downstream signaling pathways and lead to strategies for manipulating Isoform 2 levels and thus improving T cell function.

描述（由申请人提供）：IL12R¿1？TM（异构体2）是暴露于结核分枝杆菌（Mtb）后由小鼠白细胞表达的另一种il - 12受体异构体。Isoform 2缺乏IL12R¿1的跨膜结构域（Isoform 1），具有与Isoform 1不同的定位模式，并且与最初的预测相反，在体外增强了il12依赖的活性。虽然很明显，Mtb对人白细胞的激活也通过选择性剪接/聚腺苷酸化刺激异构体2的产生，但这是一个空白

项目成果

The introduction of RNA-DNA differences underlies interindividual variation in the human IL12RB1 mRNA repertoire.

RNA-DNA 差异的引入是人类 IL12RB1 mRNA 库个体间变异的基础。

• DOI: 10.1073/pnas.1515978112
• 发表时间: 2015
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Turner,AmyJ;Aggarwal,Praful;Miller,HalliE;Waukau,Jill;Routes,JohnM;Broeckel,Ulrich;Robinson,RichardT
• 通讯作者: Robinson,RichardT

IL12Rβ1: the cytokine receptor that we used to know.

• DOI: 10.1016/j.cyto.2014.11.018
• 发表时间: 2015-02
• 期刊: CYTOKINE
• 影响因子: 3.8
• 作者: Robinson, Richard T.