miRNA profiling of primate cervicovaginal lavage and extracellular vesicles reveals miR-186-5p as a potential retroviral restriction factor in macrophages

• Published in: bioRxiv
• Main Authors: Zhao, Zezhou, Muth, Dillon C, Mulka, Kathleen, Powell, Bonita H, Hancock, Grace V, Liao, Zhaohao, Metcalf Pate, Kelly A, Witwer, Kenneth W
• Format: Paper
• Language: English
• Published: Cold Spring Harbor Cold Spring Harbor Laboratory Press 31.10.2019; Cold Spring Harbor Laboratory
• Edition: 1.2
• Subjects: Antiretroviral drugs; Extracellular vesicles; HIV; Human immunodeficiency virus; Infections; Macrophages; Menopause; Menstrual cycle; Menstruation; miRNA; Molecular Biology; Monocytes; Peripheral blood; Progesterone; Replication; Reproductive system; Ultracentrifugation; HIV-1; SIV; microvesicle; cervicovaginal lavage; restriction factor; microRNA; biomarker; extracellular vesicle; exosome
• ISSN: 2692-8205; 2692-8205
• DOI: 10.1101/263947

The goal of this study was to characterize extracellular vesicles (EVs) and miRNAs of primate cervicovaginal lavage (CVL) during the menstrual cycle and simian immunodeficiency virus (SIV) infection, and to determine if differentially regulated CVL miRNAs might influence retrovirus replication. CVL and peripheral blood were collected from SIV-infected and uninfected macaques. EVs were enriched by stepped ultracentrifugation and characterized thoroughly. miRNA profiles were assessed with a medium-throughput stem-loop/hydrolysis probe qPCR platform and validated by single qPCR assays. Hormone cycling was abnormal in infected subjects, but EV concentration correlated with progesterone concentration in uninfected subjects. miRNAs were present predominantly in the EV-depleted CVL supernatant. Only a small number of CVL miRNAs were found to vary during the menstrual cycle or SIV infection. Among them was miR-186-5p, which was depleted in retroviral infection. In experiments with infected macrophages in vitro, this miRNA inhibited HIV replication. These results provide further evidence for the potential of EVs and small RNAs as biomarkers or effectors of disease processes in the reproductive tract. Footnotes * Additional analyses and experiments have been performed since the first version of this manuscript was deposited. In particular, additional EV characterization has been done, and experiments with miR-186 modulation have used cells from additional donors. These results suggest that antiretroviral efficacy of miR-186 depends on abundance/efficiency of delivery.