Bei Jiani, Qiu Yuan, Cockrell Diane, Chang Qing, Husseinzadeh Sorosh, Zhou Changcheng, Gaitas Angelo, Fang Xiang, Jin Yang, Khanipov Kamil, Saito Tais B, Gong Bin
bioRxiv. 2023 Jan 7:2023.01.06.522907. doi: 10.1101/2023.01.06.522907.
We previously reported that microRNA (miR)23a and miR30b are selectively sorted into rickettsia-infected, endothelial cell-derived exosomes ( -ECExos). Yet, the mechanism remains unknown. The number of cases of spotted fever rickettsioses has been increasing in recent years, and infections with these bacteria cause life-threatening diseases by targeting brain and lung tissues. Therefore, the aim of the present study is to continue to dissect the molecular mechanism underlying -ECExos-induced barrier dysfunction of normal recipient microvascular endothelial cells (MECs), depending on their exosomal RNA cargos. Rickettsiae are transmitted to human hosts by the bite of an infected tick into the skin. In the present study we demonstrate that treatment with -ECExos, which were derived from spotted fever group infected human dermal MECs, induced disruptions of the paracellular adherens junctional protein VE-cadherin and breached the paracellular barrier function in recipient pulmonary MECs (PMECs) in an exosomal RNA-dependent manner. Similarly, we did not detect different levels of miRs in parent dermal MECs following rickettsial infections. However, we demonstrated that the microvasculopathy-relevant miR23a-27a-24 cluster and miR30b are selectively enriched in -ECExos. Bioinformatic analysis revealed that common sequence motifs are shared exclusively among the exosomal, selectively-enriched miR23a cluster and miR30b at different levels. Taken together, these data warrant further functional identification and characterization of a single, bipartition, or tripartition among ACA, UCA, and CAG motifs that guide recognition of microvasculopathy-relevant miR23a-27a-24 and miR30b, and subsequently results in their selective enrichments in -ECExos.
我们之前报道过，微小RNA（miR）23a和miR30b被选择性地分选到立克次体感染的、内皮细胞来源的外泌体（-ECExos）中。然而，其机制仍不清楚。近年来，斑点热立克次体病的病例数一直在增加，这些细菌感染通过靶向脑和肺组织导致危及生命的疾病。因此，本研究的目的是继续剖析-ECExos诱导正常受体微血管内皮细胞（MECs）屏障功能障碍的分子机制，这取决于它们的外泌体RNA货物。立克次体通过受感染的蜱叮咬皮肤传播给人类宿主。在本研究中，我们证明，用源自斑点热群感染的人真皮MECs的-ECExos处理，以外泌体RNA依赖的方式诱导了受体肺MECs（PMECs）中细胞旁黏附连接蛋白VE-钙黏蛋白的破坏，并破坏了细胞旁屏障功能。同样，我们在立克次体感染后的亲本真皮MECs中未检测到不同水平的miR。然而，我们证明与微血管病相关的miR23a-27a-24簇和miR30b在-ECExos中选择性富集。生物信息学分析表明，外泌体中选择性富集的miR23a簇和miR30b在不同水平上仅共享共同的序列基序。综上所述，这些数据需要进一步对指导识别与微血管病相关的miR23a-27a-24和miR30b的ACA、UCA和CAG基序中的单分区、双分区或三分区进行功能鉴定和表征，随后导致它们在-ECExos中选择性富集。
