Huang Yiyao, Abdelgawad Ahmed, Turchinovich Andrey, Queen Suzanne, Abreu Celina Monteiro, Zhu Xianming, Batish Mona, Zheng Lei, Witwer Kenneth W
Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, China.
bioRxiv. 2023 Apr 29:2023.04.01.535193. doi: 10.1101/2023.04.01.535193.
Antiretroviral treatment regimens can effectively control HIV replication and some aspects of disease progression. However, molecular events in end-organ diseases such as central nervous system (CNS) disease are not yet fully understood, and routine eradication of latent reservoirs is not yet in reach. Brain tissue-derived extracellular vesicles (bdEVs) act locally in the source tissue and may indicate molecular mechanisms in HIV CNS pathology. Regulatory RNAs from EVs have emerged as important participants in HIV disease pathogenesis. Using brain tissue and bdEVs from the simian immunodeficiency virus (SIV) model of HIV disease, we profiled messenger RNAs (mRNAs), microRNAs (miRNAs), and circular RNAs (circRNAs), seeking to identify possible networks of RNA interaction in SIV infection and neuroinflammation.
Postmortem occipital cortex tissue were collected from pigtailed macaques: uninfected controls and SIV-infected subjects (acute phase and chronic phase with or without CNS pathology). bdEVs were separated and characterized in accordance with international consensus standards. RNAs from bdEVs and source tissue were used for sequencing and qPCR to detect mRNA, miRNA, and circRNA levels.
Multiple dysregulated bdEV RNAs, including mRNAs, miRNAs, and circRNAs, were identified in acute infection and chronic infection with pathology. Most dysregulated mRNAs in bdEVs reflected dysregulation in their source tissues. These mRNAs are disproportionately involved in inflammation and immune responses, especially interferon pathways. For miRNAs, qPCR assays confirmed differential abundance of miR-19a-3p, let-7a-5p, and miR-29a-3p (acute SIV infection), and miR-146a-5p and miR-449a-5p (chronic with pathology) in bdEVs. In addition, target prediction suggested that several circRNAs that were differentially abundant in source tissue might be responsible for specific differences in small RNA levels in bdEVs during SIV infection.
RNA profiling of bdEVs and source tissues reveals potential regulatory networks in SIV infection and SIV-related CNS pathology.
抗逆转录病毒治疗方案能够有效控制HIV复制及疾病进展的某些方面。然而，诸如中枢神经系统（CNS）疾病等终末器官疾病中的分子事件尚未完全明确，且潜伏储存库的常规清除目前尚无法实现。脑组织衍生的细胞外囊泡（bdEVs）在源组织中发挥局部作用，可能提示HIV中枢神经系统病理学中的分子机制。来自细胞外囊泡的调控RNA已成为HIV疾病发病机制中的重要参与者。利用来自HIV疾病的猴免疫缺陷病毒（SIV）模型的脑组织和bdEVs，我们对信使RNA（mRNAs）、微小RNA（miRNAs）和环状RNA（circRNAs）进行了分析，旨在识别SIV感染和神经炎症中可能的RNA相互作用网络。
从猪尾猕猴收集死后枕叶皮质组织：未感染对照以及SIV感染的受试者（急性期和伴有或不伴有CNS病理学的慢性期）。bdEVs根据国际共识标准进行分离和表征。来自bdEVs和源组织的RNA用于测序和qPCR，以检测mRNA、miRNA和circRNA水平。
在急性感染和伴有病理学的慢性感染中鉴定出多种失调的bdEV RNA，包括mRNA、miRNA和circRNA。bdEVs中大多数失调的mRNA反映了其源组织中的失调情况。这些mRNA不成比例地参与炎症和免疫反应，尤其是干扰素途径。对于miRNA，qPCR分析证实bdEVs中miR-19a-3p、let-7a-5p和miR-29a-3p（急性SIV感染）以及miR-146a-5p和miR-449a-5p（伴有病理学的慢性期）丰度存在差异。此外，靶标预测表明，源组织中差异丰富的几种circRNA可能是SIV感染期间bdEVs中小RNA水平特定差异的原因。
bdEVs和源组织的RNA分析揭示了SIV感染和SIV相关CNS病理学中的潜在调控网络。
