Role of miR-181a-5p in Adenovirus 14p1 induced Acute Lung Injury

miR-181a-5p在腺病毒14p1诱导的急性肺损伤中的作用

基本信息

批准号：
10621862
负责人：
Jay R Radke
金额：
$ 14.59万
依托单位：
IDAHO VETERANS RESEARCH / EDUCATION FDN
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-13 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10621862
关键词：
3&apos Untranslated Regions Acute Lung Injury Acute Respiratory Distress Syndrome Adenovirus Infections Adenovirus Protein Adenoviruses Alveolar Macrophages Animal Model Anti-Inflammatory Agents Apoptotic Cells Data Development Disease Disease Outbreaks Etiology Experimental Models Fatality rate Future Genetic Transcription Goals Hospital Mortality Human IL8 gene In Vitro Incidence Infection Infection prevention Inflammation Inflammatory Inflammatory Response Innate Immune Response Interleukin-6 Lung Lung infections Macrophage Mediating Mediator Mesocricetus auratus MicroRNAs Modeling Molecular NF-kappa B Pathogenesis Pathogenicity Pathology Pattern Play Production Pulmonary Pathology RNA Recombinants Reporter Reporting Repression Role SARS coronavirus Serotyping Severities Small RNA TNF gene Testing Therapeutic Therapeutic Intervention Translations Variant Viral Virus Virus Diseases comparative cytokine design differential expression effective therapy experimental study genetic variant immunoregulation in vivo inflammatory modulation influenza virus strain innovation lung injury new therapeutic target novel novel strategies prototype response single-cell RNA sequencing tool transcriptome sequencing translational study virology

项目摘要

PROJECT SUMMARY There have been globally distributed, regional outbreaks of a novel strain of adenovirus (Ad) 14, Ad14p1, that induces acute lung injury similar to that observed during outbreaks of infections with other Ad serotypes (e.g., Ad 3, 4 and 7). It has not, however, previously been possible to do comparative virology and pathogenesis studies, contrasting a prototype, parental Ad strain, such as Ad14, with a single, globally distributed genetic variant, such as Ad14p1. The reasons for the increased severity of these Ad14p1 infections are poorly defined. We have shown that cells undergoing Ad-induced cytopathic effect (CPE) from infection with Ad14 elicit an immunorepressive activity of activated human alveolar macrophages by repressing IL-1b, IL-6, IL-8 and TNF-alpha. In contrast, Ad14p1 CPE corpses enhance production of these same cytokines, due to decreased expression of viral E1B 20K in Ad14p1 CPE corpses. Consistent with these in vitro observations, infection of Syrian hamsters with prototype Ad14 resulted in minimal inflammation and lung injury, whereas Ad14p1 infection induced a marked acute lung injury and ARDS-like pathology. It has been shown that apoptotic cells have altered expression of microRNA (miRNA) that can repress pro- inflammatory responses and that these miRNA from apoptotic cells can be delivered to macrophages. We postulated that a similar mechanism could be occurring with Ad14 infected cells but not cells infected with the Ad14p1 variant. To test this, we infected human cells with Ad14 or Ad14p1 and did small RNA-seq on the respective CPE corpses. The data showed that Ad14 CPE corpses are enriched in four miRNA (Ad14 miRNA), all of which have been shown to repress NF-kB dependent transcription of pro-inflammatory cytokines, compared to Ad14p1 CPE corpses. Our hypothesis is that increased expression of these specific miRNA in Ad14 CPE corpses and their transfer to responder macrophages are the mechanisms through which Ad14 CPE corpses repress pro-inflammatory responses. The studies we propose here will test those hypotheses. We will test the role of Ad14 miRNA in modulating inflammatory responses of macrophages using miRNA mimics and whether E1B 20K modulates Ad14 miRNA expression during infection. We will test the role of the most abundantly expressed Ad14 miRNA, miR-181a-5p, as the mediator of Ad14 CPE immunorepression by using 3’ UTR reporter constructs and inhibiting miR-181a- 5p with miRZips. Finally, we will do translational studies to determine the role miR-181a-5p plays in the immunopathogenesis of Ad14p1 infection in the Syrian hamster. Upon completion of this project, we expect to have an increased understanding of the role of virally induced cellular miRNAs in modulation of macrophage responses during acute lung injury in response to Ad14p1. The positive impact of our studies will be the determination of the potential of miRNAs as therapeutic interventions for ARDS.

项目概要新型腺病毒 (Ad) 14、Ad14p1 株在全球范围内发生区域性爆发，引起急性肺损伤，类似于其他 Ad 血清型感染爆发期间观察到的情况（例如，Ad 3、4 和 7），但是，以前无法进行比较病毒学和发病机制。研究将原型、亲本 Ad 菌株（例如 Ad14）与单一的、全球分布的遗传菌株进行了对比变体，例如 Ad14p1。我们已经证明，这些 Ad14p1 感染严重程度增加的原因尚不清楚。 Ad14 感染导致细胞发生 Ad 诱导的细胞病变效应 (CPE)，从而引发免疫抑制活性相比之下，Ad14p1 通过抑制 IL-1b、IL-6、IL-8 和 TNF-α 来激活人肺泡巨噬细胞。由于病毒 E1B 20K 的表达减少，CPE 尸体增强了这些相同细胞因子的产生。 Ad14p1 CPE 尸体与这些体外观察一致，用原型感染叙利亚仓鼠。 Ad14 导致轻微的炎症和肺损伤，而 Ad14p1 感染则引起明显的急性肺损伤损伤和 ARDS 样病理。研究表明，凋亡细胞改变了 microRNA (miRNA) 的表达，从而抑制 pro- 炎症反应，并且这些来自凋亡细胞的 miRNA 可以被传递到巨噬细胞。假设类似的机制可能发生在 Ad14 感染的细胞中，但不会发生在感染 Ad14 的细胞中。为了测试这一点，我们用 Ad14 或 Ad14p1 感染人类细胞，并对其进行小 RNA 测序。各自的CPE尸体数据显示Ad14 CPE尸体富含四种miRNA（Ad14 miRNA），相比之下，所有这些都已被证明可以抑制促炎细胞因子的 NF-kB 依赖性转录我们的假设是 Ad14 CPE 中这些特定 miRNA 的表达增加。尸体及其转移到应答巨噬细胞是 Ad14 CPE 尸体的机制抑制促炎症反应。我们在此提出的研究将测试这些假设，我们将测试 Ad14 miRNA 在调节中的作用。使用 miRNA 模拟物研究巨噬细胞的炎症反应以及 E1B 20K 是否调节 Ad14 miRNA 我们将测试表达最丰富的 Ad14 miRNA miR-181a-5p 的作用。通过使用 3' UTR 报告构建体并抑制 miR-181a- 作为 Ad14 CPE 免疫抑制的介质最后，我们将进行翻译研究以确定 miR-181a-5p 在 miR-181a-5p 中发挥的作用。叙利亚仓鼠 Ad14p1 感染的免疫发病机制在该项目完成后，我们期望能够对病毒诱导的细胞 miRNA 在巨噬细胞调节中的作用有了更多的了解我们的研究的积极影响将是急性肺损伤期间对 Ad14p1 的反应。确定 miRNA 作为 ARDS 治疗干预的潜力。