Different Expression Pattern of Human Cytomegalovirus-Encoded microRNAs in Circulation from Virus Latency to Reactivation

Abstract

Background: Human cytomegalovirus (HCMV) is a beta-hersvirinae that has a high latent infection rate worldwide and can cause serious consequences in immunocompromised patients when reactivation; however, the mechanism of how HCMV convert from latent to reactivation has rarely been investigated. In the present study, we aimed to perform a comprehensive analysis of the HCMV-encoded microRNA (miRNA) profile in serum of patients upon HCMV reactivation from latency and to further evaluate its clinical significance for the disease monitoring and preventing usefulness.Methods: Serum samples from 60 viremia patients and 60 age-gender matched controls were enrolled in this study for screening and validation of different expression of HCMV miRNAs. Serum concentrations of 22 known HCMV miRNAs were determined by a hydrolysis probe-based stem-loop quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay. HCMV DNA was measured by quantitative real-time PCR (qPCR) with the whole blood sample. Serum HCMV IgG and IgM were assessed using enzyme linked immunosorbent assay (ELISA). Another 47 samples from 5 patients at different time points were collected to evaluate the monitoring effectiveness and disease prediction ability of differential expression HCMV-miRNAs during the antiviral treatment. Results: The RT-qPCR analysis revealed that the serum levels of 16 of the 22 examined HCMV miRNAs were elevated in HCMV viremia patients compared with controls, and a profile of 8 HCMV miRNAs including hcmv-miR-US25-2-3p, hcmv-miR-US4-5p, hcmv-miR-US25-2-5p, hcmv-miR-US25-1-3p, hcmv-miR-US25-1, hcmv-miR-UL36, hcmv-miR-UL148D, hcmv-miR-US29-3p were markedly elevated (fold change > 2, P < 0.01). Receiver operating characteristic curve (ROC) analysis were performed on the selected HCMV-miRNAs in all of the patients and controls that enrolled in this study, and which ranged from 0.74 to 0.79 in the autoimmune patients. In addition, hcmv-miR-US25-1-3p levels were significantly correlated with HCMV DNA copies (r = 0.297，P = 0.022), and were obviously higher in the reactivation set than the latency set in the autoimmune patients, which could be a predictor for the monitoring of the antiviral treatment.Conclusions: HCMV miRNAs profile showed markedly shift-switch from latency to reactivation in circulation from HCMV infected patients and hcmv-miR-US25-1-3p may be served as a predictor for the switch upon reactivation from latency in patients suffered with autoimmune diseases.

Altered serum human cytomegalovirus microRNA levels are common and closely associated with the inflammatory status in patients with fever

“…The exact reason for the negative correlation between the serum HCMV miRNAs' levels and blood CRP contents remain unclear. One possible explanation is that HCMV under latent infection status in healthy subjects with normal immunity, however, fever patients with high CRP levels often exhibited immunocompromised status, and the HCMV can be reactive when the host organism immunity attenuate (33). Nevertheless, Further studies and direct evidences are still needed to clarify this issue.…”

Significance of serum miR‐29a in the occurrence and progression of diabetic nephropathy: A cross‐sectional study

“…However, since miR2911 (5′-GGCCGGGGGACGGGCUGGGA-3′) does not have a mammalian homologue and the measurement of its expression has high repeatability and reproducibility, it was added to each sample during the RNA extraction at a concentration of 10 nmol/L. 21 The relative concentration of serum miR-29a was based on the standard concentration of miR2911 and calculated using the comparative Cq method (2 −ΔCq ), where ΔCq was calculated by subtracting the Cq value of miR2911 from the Cq value of miR-29a. Notably, miR2911 (10 nmol/L) was not subjected to serial dilution and standard curve preparation and was only used as an internal reference to represent the relative expression of miR-29a (nmol/L).…”

“…MicroRNAs (miRNAs) constitute a group of noncoding singlestranded small RNA molecules with a length of approximately [21][22][23][24][25] nucleotides that can regulate the expression of target genes through the complete or partial complementary binding of the 3' untranslated region of the target messenger RNA. 7,8 Strikingly, human serum/plasma contains numerous stably expressed miRNAs and has the biological properties of resistance to RNase activity; thus, miR-NAs derived from peripheral blood have the potential to become noninvasive biomarkers of various diseases.…”