Distinct expression profile of HCMV encoded miRNAs in plasma from oral lichen planus patients

Abstract

BackgroundOral lichen planus (OLP) is a T cell-mediated autoimmune disease. The aetiology and molecular mechanisms of OLP remain unclear. Human cytomegalovirus (HCMV) infection is a causal factor in the development of various diseases, but the clinical relevance of HCMV in OLP has not been thoroughly investigated.MethodsIn the present study, we firstly examined twenty-three HCMV-encoded microRNA (miRNA) expression profiles in plasma from training set that including 21 OLP patients and 18 healthy controls using RT-qPCR technology. Dysregulated miRNAs were subsequently confirmed in another larger cohort refereed as validation set consisting of 40 OLP patients and 33 healthy controls. HCMV DNA in peripheral blood leukocytes (PBLs) was also measured in an additional cohort of 13 OLP patients and 12 control subjects. Furthermore, bioinformatics analyses, luciferase reporter assay and western blotting were also performed to predict and verify the direct potential targets of HCMV-encoded miRNAs.ResultsThe RT-qPCR results showed that the plasma levels of five HCMV-encoded miRNAs including hcmv-miR-UL112-3p, hcmv-miR-UL22a-5p, hcmv-miR-UL148d, hcmv-miR-UL36-5p and hcmv-miR-UL59 were significantly increased in OLP patients in both training and validation sets. HCMV DNA in PBLs was also significantly higher in OLP patients than in control subjects. Additionally, by using a combination of luciferase reporter assay and western blotting, we demonstrated that cytomegalovirus UL16-binding protein 1, a molecule that mediates the killing of virus-infected cells by natural killer cells, is a direct target of hcmv-miR-UL59.ConclusionsOur results demonstrate a distinct expression pattern of HCMV-encoded miRNAs in OLP patients, which may provide insight into the relationship between HCMV infection and OLP, and warrants additional study in the diagnosis and aetiology of OLP.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-017-1222-8) contains supplementary material, which is available to authorized users.

Altered human cytomegalovirus-encoded miRNAs in host circulation: novel disease biomarkers and potential aetiological agents

“…More interestingly, recent studies revealed that viruses can hijack the exosome pathway, and virus-encoded miRNAs can be actively secreted via exosomes from virus-infected cells and delivered to and act in virus-negative recipient cells [34]. Furthermore, our latest study implied that specifically altered plasma HCMV miRNA is primarily encapsulated in exosomes [35]. Thus, taken together our findings combined with those previous reports, the exogenous HCMV-encoded miRNAs can be packaged into exosomes and delivered to recipient cells, moreover, the involvement of exosome-encapsulated HCMV-encoded miRNAs in host recipient cellular gene repression may also provide further insights into novel and somewhat surprising potential therapeutic targets.…”

“…Oral lichen planus Upregulation [35] this disease remains elusive. By characterizing the plasma miRNAs pattern of patients with essential hypertension, Li and colleagues identified that the plasma concentrations of miR-UL112 were markedly higher in hypertensive patients than that in the control subjects.…”

The E2F1–miR-520/372/373–SPOP Axis Modulates Progression of Renal Carcinoma

“…Protein was extracted from tissues and cells by RIPA lysis buffer, and protein levels were analyzed via Western blotting and performed as described previously (26). The antibodies used for Western blotting were as follows: anti-SPOP antibody (ProteinTech; 16750-1-AP, 1:500), anti-E2F1 antibody (Cell Signaling Technology; #3742, 1:1,000), anti-PTEN antibody (Cell Signaling Technology; #9559, 1:1,000), anti-DUSP7 antibody (ABGENT; AP8450a, 1:500), anti-SP1 antibody (Abcam; ab227383,1:1,000), anti-c-Myc antibody (Cell Signaling Technology; #13987, 1:1,000), and anti-GAPDH antibody (Santa Cruz Biotechnology; sc-25778, 1:2,000).…”

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

“…Comparatively, detection of hcmv-miR-UL22A-5p in transplant recipients with HCMV infections had the highest sensitivity for the prediction of subsequent virologic recurrence [16]. Our research group also demonstrated different expression pro le of HCMVencoded miRNAs in plasma sample from patients suffered with OLP, and 5 of the miRNAs including hcmv-miR-UL112-3p, hcmv-miR-UL22a-5p, hcmv-miR-UL148D, hcmv-miR-UL36-5p and hcmv-miR-UL59 were signi cantly upregulated in OLP samples compared with normal samples [17]. All these results indicate that these HCMV-encoded-miRNAs may share similar physiological and pathological roles in HCMV infection related diseases mentioned above, as well as in this study, even though much further study is needed to con rm this.…”

“…Total RNA was extracted from 100 µL of serum using a 1-step phenol/chloroform puri cation method and precipitated using isopropyl alcohol as previously described [17]. In brief, 100 µL of serum was mixed with 300 µL deionized water, 200 µL acid phenol, and 200 µL chloroform.…”