Table_1_ERG-Associated lncRNA (ERGAL) Promotes the Stability and Integrity of Vascular Endothelial Barrier During Dengue Viral Infection via Interaction With miR-183-5p.DOCX

Dengue virus (DENV) continues to be a major public health problem. DENV infection will cause mild dengue and severe dengue. Severe dengue is clinically manifested as serious complications, including dengue hemorrhagic fever and/or dengue shock syndrome (DHF/DSS), which is mainly characterized by vascular leakage. Currently, the pathogenesis of severe dengue is not elucidated thoroughly, and there are no known therapeutic targets for controlling the disease effectively. This study aimed to further reveal the potential molecular mechanism of severe dengue. In this study, the long non-coding RNA, ERG-associated lncRNA (lncRNA-ERGAL), was activated and significantly up-regulated in DENV-infected vascular endothelial cells. After knockdown of lncRNA-ERGAL, the expression of ERG, VE-cadherin, and claudin-5 was repressed; besides, cell apoptosis was enhanced, and cytoskeletal remodeling was disordered, leading to instability and increased permeability of vascular endothelial barrier during DENV infection. Fluorescence in situ hybridization (FISH) assay showed lncRNA-ERGAL to be mainly expressed in the cytoplasm. Moreover, the expression of miR-183-5p was found to increase during DENV infection and revealed to regulate ERG, junction-associated proteins, and the cytoskeletal structure after overexpression and knockdown. Then, ERGAL was confirmed to interact with miR-183-5p by luciferase reporter assay. Collectively, ERGAL acted as a miRNA sponge that can promote stability and integrity of vascular endothelial barrier during DENV infection via binding to miR-183-5p, thus revealing the potential molecular mechanism of severe dengue and providing a foundation for a promising clinical target in the future.

登革热病毒（DENV）持续构成重大的公共卫生问题。DENV感染可导致轻微的登革热和严重的登革热。严重的登革热在临床上表现为严重的并发症，包括登革出血热和/或登革休克综合征（DHF/DSS），其主要特征为血管渗漏。目前，严重登革热的发病机制尚未得到充分阐明，且尚无已知的有效治疗靶点。本研究旨在进一步揭示严重登革热潜在的分子机制。在本研究中，长非编码RNA，ERG相关长链非编码RNA（lncRNA-ERGAL），在DENV感染血管内皮细胞中被激活并显著上调。通过敲低lncRNA-ERGAL，ERG、VE-钙粘蛋白和claudin-5的表达受到抑制；此外，细胞凋亡增强，细胞骨架重塑紊乱，导致DENV感染期间血管内皮屏障的不稳定性和通透性增加。荧光原位杂交（FISH）实验显示lncRNA-ERGAL主要在细胞质中表达。此外，miR-183-5p的表达在DENV感染期间被发现增加，并在过表达和敲低后调节ERG、连接蛋白和细胞骨架结构。随后，通过荧光素酶报告基因实验证实ERGAL与miR-183-5p相互作用。综上所述，ERGAL作为一种miRNA海绵，通过结合miR-183-5p，能够促进DENV感染期间血管内皮屏障的稳定性和完整性，从而揭示了严重登革热的潜在分子机制，并为未来潜在的临床治疗靶点奠定了基础。