Molecular pathology of oral immune dysregulation in HIV/SIV infection

摘要 虽然，在急性HIV/SIV感染期间，口腔粘膜中维持了先天性和适应性免疫功能， 感染后，这些反应在慢性HIV感染期间明显受损， 联合抗逆转录病毒治疗（cART）。持续性口腔粘膜炎症/免疫激活 以促炎性细胞因子产生失调和口腔生态失调为特征的口腔溃疡的主要特征， 慢性HIV/SIV感染。尽管我们对HIV/SIV感染引起的口腔粘膜功能障碍的理解 尽管这些基因的表达水平得到了改善，但其潜在的分子机制仍然是未知的和未探索的。除了细胞因子和 转录因子，最近的研究表明microRNA（miRNA）介导的基因调控是至关重要的 免疫细胞分化、成熟、活化和炎症反应。在其他领域的广泛研究 口腔的慢性炎性疾病（牙周炎、口腔扁平苔藓。等等）。揭示了一种 miRNAs在疾病发病机制中的致病作用导致其被鉴定为有希望的治疗药物 目标的然而，miRNA在HIV诱导的口腔粘膜功能障碍中的作用缺乏。本申请 建立在初步研究的基础上，该研究确定了与T和B细胞相关的miRNAs的显著失调， 慢性SIV感染的扁桃体和颊粘膜（BUCM）中的激活和上皮屏障破坏 恒河猴使用δ-9-四氢大麻酚（δ-9-tetrahydrocannabinol，THC）给药的同一组研究 慢性SIV感染的RM显著抑制胃肠道促炎基因/miRNA表达 并在体外抑制T和B细胞的活化和增殖，表明其具有巨大的治疗潜力 用于减弱/逆转局部（口腔）和全身免疫激活并减缓疾病进展， 抑制性cART的设置。基于我们强有力的初步数据表明， 在慢性HIV/SIV感染的BUCM和扁桃体中表达失调，我们假设这导致了口腔 粘膜功能障碍，这有助于免疫激活和HIV疾病进展。我们还 假设慢性四氢大麻酚-THC治疗可以减少炎症，恢复口腔免疫功能， HIV/SIV疾病进展通过调节miRNA表达。此应用程序的一个主要目标是识别 与口腔粘膜功能障碍相关的miRNA机制和大麻素介导的抑制 免疫激活该项目有三个具体目标：1）验证miRNA（miR-142- 3 p，-142- 3 p） 5 p，-186，-7，-101- 3 p，-29 b，-141）介导的闭合蛋白（occludin，OCLN）、封闭蛋白-1（claudin-1，CLDN 1）和封闭蛋白-1（claudin-1，CLDN 2）的下调。 封闭齿（ZO 1）导致口腔上皮屏障破坏和上皮渗透性增加。2)识别 大麻素诱导的BUCM和扁桃体中免疫激活抑制的miRNA机制 SIV感染的过程。3)确定联合抗逆转录病毒治疗（cART）的效果 在慢性给药的情况下，对口腔微生物菌群、炎症/免疫激活、病毒复制 口腔上皮细胞紧密连接蛋白与miRNA的表达这项研究是新颖的， 应用最先进的免疫学和分子方法来解决我们的一个重大差距， 理解口腔粘膜功能障碍的转录和转录后机制 在抑制性cART的情况下。此外，更重要的是，这些结果将加深我们的 了解大麻素抗炎作用的机制， 不仅对HIV而且对其他慢性炎症性疾病的靶向免疫调节的治疗意义 口腔疾病。因此，总体意义和影响很大。

ABSTRACT Although, innate and adaptive immune functions are maintained in the oral mucosa during acute HIV/SIV infection, these responses become markedly impaired during chronic HIV infection and are not fully restored by combination anti-retroviral therapy (cART). Persistent oral mucosal inflammation/immune activation characterized by dysregulated proinflammatory cytokine production and oral dysbiosis are cardinal features of chronic HIV/SIV infection. Even though our understanding of oral mucosal dysfunction in HIV/SIV infection has improved, the underlying molecular mechanisms remain unknown and unexplored. Apart from cytokines and transcription factors, recent studies show microRNA (miRNA)-mediated gene regulation to be critical for immune cell differentiation, maturation, activation and inflammatory responses. Extensive studies in other chronic inflammatory diseases of the oral cavity (Periodontitis, Oral lichen planus. etc.) have revealed a causative role for miRNAs in disease pathogenesis resulting in their identification as promising therapeutic targets. However, the role of miRNAs in HIV induced oral mucosal dysfunction is lacking. This application builds on preliminary studies, which identified significant dysregulation of miRNAs linked to T and B-cell activation and epithelial barrier disruption in tonsils and buccal mucosa (BUCM) from chronically SIV-infected rhesus macaques. Studies of the same group using delta-9-tetrahydrocannabinol (⁹-THC) administration to chronic SIV-infected RMs significantly suppressed gastrointestinal proinflammatory gene/miRNA expression and inhibited T and B-cell activation and proliferation in vitro, suggesting their immense therapeutic potential for attenuating/reversing local (oral cavity) and systemic immune activation and slowing disease progression in the setting of suppressive cART. Based on our strong preliminary data demonstrating that miRNA expression is dysregulated in BUCM and tonsils in chronic HIV/SIV infection, we hypothesize that this leads to oral mucosal dysfunction, which contributes to immune activation and HIV disease progression. Further, we hypothesize that chronic ⁹-THC treatment may reduce inflammation, restore oral immune function, and slow HIV/SIV disease progression by modulating miRNA expression. A major goal of this application is to identify miRNA mechanisms associated with oral mucosal dysfunction and cannabinoid mediated suppression of immune activation. The project has three specific aims: 1) Test the hypothesis that miRNA (miR-142-3p,-142- 5p,-186,-7,-101-3p,-29b,-141) mediated downregulation of occludin (OCLN), claudin-1 (CLDN1) and zona occludens (ZO1) leads to oral epithelial barrier breakdown and increased epithelial permeability. 2) Identify miRNA mechanisms of cannabinoid-induced suppression of immune activation in BUCM and tonsils during the course of SIV infection. 3) Determine the effect of combination anti-retroviral treatment (cART) in conjunction with chronic ⁹-THC administration on the oral microflora, inflammation/immune activation, viral replication, miRNA expression and epithelial tight junction proteins in the oral cavity. The proposed research is novel and applies state of the art immunological and molecular approaches to address a significant gap in our understanding of the transcriptional and post-transcriptional mechanisms underlying oral mucosal dysfunction in the setting of suppressive cART. Additionally and more importantly, the results will deepen our understanding of the mechanisms underlying the anti-inflammatory effects of cannabinoids and have important therapeutic implications for targeted immune modulation in not only HIV but also other chronic inflammatory diseases of the oral cavity. Thus, the overall significance and impact is high.