Molecular mechanisms of posttranscriptional gene regulation in asthmatic airway inflammation

哮喘气道炎症转录后基因调控的分子机制

• 批准号: 10698606
• 负责人: ULUS ATASOY
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10698606
• 关键词: Ablation; Allergens; Asthma; Binding; Biological Products; CD4 Positive T Lymphocytes; Cell physiology; Chemicals; Data; Development; Disease; Donor person; Elements; Family; Family member; GATA3 gene; Gene Cluster; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Genetically Engineered Mouse; Goals; Healthcare Systems; Human; Immunoprecipitation; Inflammation; Inflammatory; Intervention; Investigation; Knock-in; Knowledge; Lung; Lung Transplantation; Mediating; Messenger RNA; Methods; MicroRNAs; Military Personnel; Modeling; Molecular; Mus; Oral; Outcome; Paint; Pathogenesis; Patients; Peripheral; Pharmaceutical Preparations; Play; Post-Transcriptional Regulation; Production; Proteins; Publishing; Pulmonary Inflammation; RNA; RNA-Binding Proteins; Regulation; Research; Role; Steroid Resistance; Steroid-resistant asthma; Steroids; T cell differentiation; T-Lymphocyte; TIS11 protein; Techniques; Testing; Therapeutic; Trans-Activators; Transcript; Treatment outcome; Veterans; airway inflammation; allergic airway inflammation; asthmatic; asthmatic airway; cytokine; experimental study; genetic signature; insight; mRNA Expression; mRNA Stability; mRNA Translation; member; military veteran; mouse model; novel; novel therapeutics; overexpression; peripheral blood; permissiveness; posttranscriptional; response; side effect; small hairpin RNA; small molecular inhibitor; transcriptome sequencing; transcriptomics; treatment response

Asthma remains a difficult to treat disease that greatly impacts deployed military personnel and Veterans. Currently used medications either: 1) are very expensive (biologics), placing enormous burdens on the VA Healthcare System; 2) have significant side effects (oral steroids); or 3) do not work in some endotypes, e.g., steroid-resistant asthma. Due to discordance between steady-state mRNA levels and protein, transcriptomic approaches may overlook genes regulated by RNA binding proteins (RBPs). Posttranscriptional gene regulation by RBPs and microRNAs (miRNAs) is increasingly recognized as an important control mechanism for pro- inflammatory genes but understudied. RBPs, such as HuR (Elavl1), which binds to mRNA AU-rich elements (AREs), play critical roles by regulating mRNA stability and translation of key pro-inflammatory gene expression in asthma. Our recently published data indicates that HuR ablation in mice ameliorates allergen-driven lung inflammation. Furthermore, HuR is over-expressed in asthmatic CD4+ T cells and its inhibition reduces cytokine expression. Using RNA Immunoprecipitation techniques (RIP-seq) combined with genetically engineered murine models, we have demonstrated that HuR controls both Th2 and Th17 CD4+ T lineages. Without a better understanding of posttranscriptional control of inflammation, the field will continue to have a limited insight into molecular mechanisms, which likely contribute to asthma endotypes and unequal treatment responses and outcomes in patients. Our long-term goal is to understand posttranscriptional gene regulation in different endotypes of asthmatic airway inflammation. The objective of this application is to determine how HuR and TTP family members regulate key pro-inflammatory molecules produced by CD4+ T cells in different asthma endotypes. Our rationale is that investigation of HuR-driven gene expression will identify molecular mechanisms that differ between asthma endotypes, especially type 2-high vs. non-type 2-high. The central hypothesis is that the HuR-Gata3 interaction in CD4+ T cells controls airway inflammation in type 2-high asthma (driven by Gata3) and in non-type 2-high asthma (driven by Il17). We plan to test the central hypothesis and accomplish these objectives by the following three specific aims: 1) Define molecular mechanisms of HuR regulation in murine models of type 2-high airway inflammation; 2) Elucidate human CD4+ T cell gene clusters permissive for asthmatic lung inflammation and 3) Determine effects of HuR inhibition on T cell-mediated inflammation in type 2 high and non-type 2 high asthma. At the completion of the proposed research, our expected outcomes are to identify how HuR controls CD4+ T cell differentiation and function, which are critical for the development of allergic airway inflammation. These results are anticipated to have a sustained positive impact upon the field because they will further define molecular mechanisms distinguishing type 2 high from non-type 2 high asthma endotypes. The fundamentally important knowledge gained will provide opportunities to develop novel therapies to treat asthmatic lung inflammation by interfering with HuR function, including in steroid-resistant asthma.

哮喘仍然很难治疗极大地影响部署的军事人员和退伍军人的疾病。 目前使用的药物要么：1）非常昂贵（生物制剂），在VA上施加了巨大的负担 医疗保健系统； 2）具有显着的副作用（口服类固醇）；或3）在某些内型中不起作用，例如 抗类固醇的哮喘。由于稳态mRNA水平和蛋白质之间的不一致，转录组 方法可能会忽略由RNA结合蛋白（RBP）调节的基因。转录后基因调控 RBP和MicroRNA（miRNA）越来越被认为是重要的控制机制 炎症基因但研究了。 RBP，例如HUR（Elavl1），它与mRNA au-rich元素结合 （ARES），通过调节mRNA稳定性和关键促炎基因表达的翻译来发挥关键作用 在哮喘中。我们最近发表的数据表明，小鼠的HUR消融可以改善过敏原驱动的肺 炎。此外，HUR在哮喘CD4+ T细胞中过度表达及其抑制作用可降低细胞因子 表达。使用RNA免疫沉淀技术（RIP-SEQ）结合了基因工程的鼠 模型，我们证明了HUR控制TH2和TH17 CD4+ T谱系。没有更好的 了解炎症后转录后控制，该领域将继续对 分子机制，可能有助于哮喘内型和不平等的治疗反应以及 患者的结果。我们的长期目标是了解不同的转录后基因调节 哮喘气道炎症的内型。该应用的目的是确定HUR和TTP如何 家庭成员调节CD4+ T细胞在不同哮喘中产生的关键促炎分子 内型。我们的理由是，对HUR驱动基因表达的研究将识别分子机制 哮喘内型，尤其是2高的哮喘内型与非型2高的哮喘内型之间有所不同。中心假设是 CD4+ T细胞中的HUR-GATA3相互作用控制2型哮喘的气道炎症（由 GATA3）和非型2高哮喘（由IL17驱动）。我们计划测试中心假设并完成 这些目标通过以下三个特定目的：1）定义HUR调节的分子机制 2型高气道炎症的鼠模型； 2）阐明人类CD4+ T细胞基因簇允许 哮喘性肺部炎症和3）确定HUR抑制对T细胞介导的炎症类型的影响 2高和非型2高哮喘。拟议研究完成时，我们的预期结果是 确定HUR如何控制CD4+ T细胞分化和功能，这对于开发至关重要 过敏性气道炎症。预计这些结果会对现场产生持续的积极影响 因为它们将进一步定义分子机制，从而区分2型高的非类型2高哮喘 内型。从根本上获得的重要知识将提供开发新疗法的机会 通过干扰HUR功能，包括抗类固醇的哮喘。