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.

哮喘仍然是一种难以治疗的疾病，极大地影响了已部署的军事人员和退伍军人。 目前使用的药物有：1) 非常昂贵（生物制剂），给 VA 带来巨大负担 医疗保健系统； 2）有明显的副作用（口服类固醇）；或 3) 在某些内型中不起作用，例如， 类固醇抵抗性哮喘。由于稳态 mRNA 水平和蛋白质之间的不一致，转录组学 方法可能会忽略 RNA 结合蛋白 (RBP) 调控的基因。转录后基因调控 RBP 和 microRNA (miRNA) 被越来越多地认为是一种重要的控制机制。 炎症基因但尚未得到充分研究。 RBP，例如 HuR (Elavl1)，可与富含 mRNA AU 的元件结合 (ARE)，通过调节 mRNA 稳定性和关键促炎基因表达的翻译发挥关键作用 在哮喘中。我们最近发表的数据表明，小鼠中的 HuR 消融可改善过敏原驱动的肺部 炎。此外，HuR 在哮喘 CD4+ T 细胞中过度表达，其抑制可减少细胞因子 表达。使用RNA免疫沉淀技术（RIP-seq）结合基因工程小鼠 在模型中，我们证明了 HuR 控制 Th2 和 Th17 CD4+ T 谱系。没有更好的 尽管对炎症转录后控制的了解，该领域仍将有有限的了解 分子机制，可能导致哮喘内型和不平等的治疗反应， 患者的结果。我们的长期目标是了解不同物种中的转录后基因调控 哮喘气道炎症的内型。此应用程序的目的是确定 HuR 和 TTP 如何 家族成员在不同哮喘中调节 CD4+ T 细胞产生的关键促炎分子 内型。我们的理由是，对 HuR 驱动的基因表达的研究将确定分子机制 哮喘内型之间存在差异，尤其是 2 型高与非 2 型高。中心假设是 CD4+ T 细胞中的 HuR-Gata3 相互作用控制 2 型高哮喘中的气道炎症（由 Gata3）和非 2 型高哮喘（由 Il17 驱动）。我们计划检验中心假设并完成 这些目标通过以下三个具体目标：1）定义HuR调节的分子机制 2型高气道炎症小鼠模型； 2) 阐明人类 CD4+ T 细胞基因簇 哮喘性肺部炎症和 3) 确定 HuR 抑制对 T 细胞介导的炎症的影响 2型高和非2型高哮喘。完成拟议的研究后，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 CD4+ T 细胞的发育至关重要 过敏性气道炎症。预计这些结果将对该领域产生持续的积极影响 因为他们将进一步定义区分 2 型高哮喘和非 2 型高哮喘的分子机制 内型。获得的基本重要知识将为开发新疗法提供机会 通过干扰 HuR 功能来治疗哮喘性肺部炎症，包括类固醇耐药性哮喘。