A LncRNA in Pulmonary Mucosal Immunity and HIV-Associated Comorbidities

项目摘要 当代抗逆转录病毒疗法已将艾滋病毒感染转化为医学上可控制的慢性疾病， 条件然而，艾滋病毒感染者（PLWH）的寿命更长，使他们更容易感染艾滋病毒。 其他感染和共病。在调整潜在混杂因素后，PLWH比正常人高50-60%， 有可能发展为慢性阻塞性肺病（COPD），而且也是在更年轻的时候。 吸烟在艾滋病毒感染者中非常普遍，吸烟者艾滋病毒感染者（SLWH）表现出早发性 慢性阻塞性肺疾病患者肺功能加速下降。此外，艾滋病毒也是肺部感染的独立危险因素。 病理学HIV改变了包括肺在内的多个器官的上皮细胞内稳态;然而， HIV对肺上皮炎症反应的影响还不清楚。以前，使用大型动物模型， 我们报告了HIV和CS之间在影响肺部病理生理学方面存在强烈的相互作用。我们的数据表明 HIV是引起上皮屏障功能障碍和慢性肺部病变的一个强有力的独立危险因素， 和CS-暴露协同加重肺上皮重塑。尽管如此， 关于导致SLWH这种情况的宿主肺因素。分析肺癌的遗传决定因素 上皮反应，进行RNA测序，并鉴定长非编码RNA（lncRNA） 与上皮炎症反应显著相关。基于实验验证 以及在基因调控和调节炎症方面的潜力，LncRNA反义ICAM-1（LASI）是 选择进行进一步分析。细胞内粘附分子1（ICAM-1）是一种细胞表面糖蛋白， 在上皮细胞中，包括支气管上皮细胞，与气道炎症相关， 传播病毒感染到其他细胞并导致对慢性疾病的易感性增加。LASI 与ICAM-1重叠，在反义链上编码，在CS处理的正常人中高度上调。 人气道上皮细胞（HAEC）和COPD受试者与对照组相比。此外，击倒 HAECs中的这种lncRNA抑制炎性因子。因此，这些数据表明lncRNA LASI可能是 在HIV感染的背景下，是气道上皮反应和COPD发病机制的重要调节剂。 因此，我们认为HIV感染和CS暴露改变了lncRNA LASI表达和亚细胞 在气道中的定位导致宿主对气道炎症和COPD恶化的易感性。我们将 通过1）通过功能的获得和丧失直接调节气道上皮细胞中的LASI水平， 研究; 2）通过分析LASI调节炎症反应的机制， 作为microRNA或微蛋白的前体，或作为结合microRNA的海绵。的 因此，拟议的研究将有助于提高我们对环境暴露（如CS）影响的理解 和艾滋病毒在肺上皮细胞，并可能提供新的见解，为恢复和改善肺， SLWH的整体健康状况。

PROJECT ABSTRACT Contemporary antiretroviral therapy has transformed HIV infection into a medically manageable chronic condition. However, the longer lifespan of people living with HIV (PLWH) has made them more susceptible to other infections and comorbid diseases. After adjusting for potential confounders, the PLWH are 50-60% more likely to develop the chronic obstructive pulmonary disease (COPD) and that too at a much younger age. Cigarette smoking is highly prevalent among PLWH and smokers living with HIV (SLWH) exhibit an early onset of COPD with an accelerated decline in lung functions. Moreover, HIV is also an independent risk factor for lung pathologies. HIV alters the epithelial homeostasis in multiple organs, including lungs; however, the cellular impact of HIV on lung epithelial inflammatory responses is poorly defined. Previously, using a large animal model, we reported a strong interaction between HIV and CS in affecting the lung pathophysiology. Our data suggested that HIV is a strong independent risk factor in causing epithelial barrier dysfunction and chronic lung pathologies, and CS-exposure synergistically exacerbated the lung epithelial remodeling. Nonetheless, not much is known about the host lung factors contributing to this condition in SLWH. To analyze the genetic determinants of lung epithelial responses, RNA sequencing was performed, and the long noncoding RNAs (lncRNAs) were identified that were significantly associated with epithelial inflammatory responses. Based on the experimental validation and the potential in gene regulation and in modulating inflammation, LncRNA Anti-Sense to ICAM-1 (LASI) was selected for further analyses. Intracellular adhesion molecule 1 (ICAM-1) is a cell surface glycoprotein expressed in epithelial cells, including bronchial epithelia and is associated with airway inflammation and potentially propagates viral infection to other cells and lead to increased susceptibility to chronic diseases. The LASI overlaps with ICAM-1 and is encoded on the antisense strand and was highly upregulated in CS treated normal human airway epithelial cells (HAECs) and in COPD subjects compared to controls. Furthermore, knocking down this lncRNA in HAECs suppressed the inflammatory factors. These data thus posit that lncRNA LASI could be an essential modulator of airway epithelial response and COPD pathogenesis in the context of HIV infection. Therefore, we propose that HIV infection and CS exposure alters the lncRNA LASI expression and subcellular localization in airways resulting in host susceptibility to airway inflammation and COPD exacerbations. We will test this hypothesis by 1) directly modulating the LASI levels in airway epithelial cells by gain- and loss-of-function studies; 2) identifying the mechanisms by which LASI modulates inflammatory responses by analyzing its potential as the precursor for microRNAs or microproteins, or as the sponge that binds the microRNAs. The proposed studies will thus help in improving our understanding of the effect of environmental exposures like CS and HIV in lung epithelial cells and may provide new insights for restoring and improving the pulmonary and overall health of SLWH.