486Serum levels of miRNA-221 and -222 may be predictive biomarkers for cognitive decline

Background Although dementia is a huge problem in public health, no fundamental biomarker has been established to detect cognitive decline at the early stage. MicroRNAs (miRNAs) regulate gene expression, and are associated with the development of various diseases. Methods The subjects of this prospective study were 162 (75 men, 87 women) residents who attended a health examination in Yakumo town in Hokkaido, in 2012 and re-attended at least once while 2013 to 2015. Serum samples were collected in 2012 and serum miRNA were measured by qRT-PCR. We used a short version of the Mini-Mental State Examination (SMMSE) to screen cognitive function, and calculated the change in SMMSE score per year. We calculated odds ratios (ORs) and 95% confidence intervals (CIs) of serum miRNA levels for cognitive decline (decrease of greater than 0.5 points per year) using the lowest tertile group of miRNAs as the reference by logistic regression analysis. Results The mean age and change in SMMSE score of the subjects was 63.9±9.6 years and -0.03±1.19 points. Odds ratios (ORs) for cognitive decline were significantly higher in the highest tertile of serum miR-221 (OR = 3.24, 95%CI=1.20-8.72) and miR-222 (OR = 4.01, 95%CI=1.36-11.80) even if confounding factors were adjusted. Conclusions High serum levels of miR-221 and -222 were significantly associated with cognitive decline. Key messages High serum levels of miR-221 and -222 may be potential candidate biomarkers for prediction of cognitive decline.

Circulating miRNA expression in asthmatics is age-related and associated with clinical asthma parameters, respiratory function and systemic inflammation

Background The course of asthma may differ between elderly asthmatics (EA) and non-elderly asthmatics (nEA), which may be partially associated with an age-dependent aberrant immune response. The aim of the study was to determine the influence of serum miRNA expression on asthma characteristics and systemic inflammation markers in EA and nEA. Methods Control and severity of asthma, pulmonary function and FeNO were assessed in 28 EA and 31 nEA patients. The control group included 59 elderly and non-elderly healthy individuals. The expression of selected miRNAs in serum was measured with rt-PCR, and proinflammatory cytokine activity was assayed by ELISA or flow cytometry. Results No difference in serum miRNA expression was observed between the asthmatics and healthy controls. EA demonstrated lower expression of miRNA-106a and miRNA-126a than nEA (p = 0.003 and p = 0.02) and EC had lower expression of miRNA-146a, -126a, -106a and 19b than nEC (p = 0.001, p = 0.003, p = 0.005 and p < 0.001 respectively). Only nEA demonstrated a relationship between the expression of selected miRNAs and the level of asthma control (assessed with ACT) and with airway inflammation, measured by FeNO level. All patients with asthma demonstrated elevated TNFα, IL-6 and sTNF RI levels compared to controls (p = 0.026, p = 0.03 and p < 0.001 respectively). EA demonstrated a higher TNFα level than EC (p < 0.001), and EA had a higher level of sTNF RI than nEA (p < 0.001). A significant correlation was observed between serum levels of proinflammatory cytokines and selected miRNAs. Conclusion Serum miRNA expression was found to correlate with clinical characteristics of asthma and systemic inflammation in an age-dependent fashion, suggesting that miRNA may differentially contribute to asthma pathogenesis in elderly and non-elderly patients.

Serum miRNAs Support the Indication for MRI-Ultrasound Fusion-Guided Biopsy of the Prostate in Patients with Low-PI-RADS Lesions

Multiparametric MRI (mpMRI) and targeted biopsy of the prostate enhance the tumor detection rate. However, the prediction of clinically significant prostate cancer (PCa) is still limited. Our study tested the additional value of serum levels of selected miRNAs in combination with clinical and mpMRI information for PCa prediction and classification. A total of 289 patients underwent targeted mpMRI-ultrasound fusion-guided prostate biopsy complemented by systematic biopsy. Serum miRNA levels of miRNAs (miR-141, miR-375, miR-21-5p, miR-320b, miR-210-3p, let-7c, and miR-486) were determined by quantitative PCR. Detection of any PCa and of significant PCa were the outcome variables. The patient age, pre-biopsy PSA level, previous biopsy procedure, PI-RADS score, and serum miRNA levels were covariates for regularized binary logistic regression models. The addition of miRNA expression of miR-486 and let-7c to the baseline model, containing only clinical parameters, increased the predictive accuracy. Particularly in patients with PI-RADS ≤3, we determined a sensitivity for detecting significant PCa (Gleason score≥7a corresponding to Grade group ≥2) of 95.2%, and an NPV for absence of significant PCa of 97.1%. This accuracy could be useful to support patient counseling in selected cases.

Changes in Expression of miRNA-320a and miRNA-497-5p in Early Stage of Breast Cancer

Background: Micro-ribonucleic acids (miRNAs) are noncoding small RNA species considered a varying class with a single-stranded structure whose expression is often dysregulated in cancer. The expression of miRNAs has been used as a promising new biomarker for the detection of breast cancer (BC). Objectives: The purpose of the present case-control study was to investigate the expression levels of miRNA-320a and miRNA-497-5p and their potential role in BC patients in comparison to those of the healthy controls in the Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran, in 2019. Methods: The concentrations of miR-320a and miR-497-5p were analyzed in 80 serum samples of 40 patients with a confirmed diagnosis of early-stage BC in comparison to those of 40 age-matched healthy volunteers. Real-time quantitative polymerase chain reaction was carried out for the detection of the expression level of these miRNAs. Results: The results of the current study showed that the serum levels of miR-320a and miR-497-5p were down-regulated in the BC patients, compared to those reported for the healthy controls (P=0.651 and P=0.044, respectively). However, the levels of miR-320a in the early-stage BC samples were not statistically different from those of the healthy volunteers. There was a reduction in the serum miRNA-320a of the premenopausal subjects under 48 years of age. Serum miRNA-497-5p also decreased among the cases under 48 years of age. Conclusions: The identification and effectiveness of these miRNAs were demonstrated in the early-stage BC screening. It seems that miRNAs have the potential to be used as biomarkers for the screening and diagnosis of BC.

MO635PRO-INFLAMMATORY CYTOKINES IL-6 AND IL-17 DISPLAY A PARTICULAR MOLECULAR PATTERN IN ASSOCIATION WITH DYSREGULATED MIRNAS IN PATIENTS WITH TYPE 2 DIABETES MELLITUS IN THE EARLY STAGES OF DIABETIC KIDNEY DISEASE

Background and Aims Glomerular injury and proximal tubule (PT) dysfunction have intricate mechanisms in diabetic kidney disease (DKD). Pro-inflammatory cytokines are involved in the initiation and progression of DKD through mediating the inflammatory response, both at glomerular and proximal tubular level. miRNAs are able to modulate cellular and biochemical functions, thus intervening in the pathogenesis of DKD. The aim of the study, performed on patients with type 2 diabetes mellitus (DM), was to evaluate selective pro-inflammatory cytokines in relation to biomarkers of podocyte lesion and of PT dysfunction. Particular molecular pathways, such as specific miRNA profiles operating in this relation, have also been studied. Method A number of 126 patients with type 2 DM, staged by albuminuria [39 normoalbuminuric – urinary albumin/creatinine ratio UACR)&lt;30mg/g; 45 microalbuminuric–UACR-30–300mg/g; 42 macroalbuminuric–UACR&gt;300mg/g], and 23 healthy control subjects were included in a cross-sectional study. All patients were evaluated concerning biomarkers of podocyte injury (nephrin, podocalyxin, synaptopodin) and of PT dysfunction [Kidney injury molecule-1(KIM-1), N-acetyl-beta-D-glucuronidase (NAG), alpha 1- microglobulin]. Also, serum and urinary levels of specific interleukins (IL-6, IL-17), serum cystatin C, and eGFR were determined. Serum and urinary miRNAs (miRNA-21, miRNA-124, miRNA-146a, miRNA-192) were assessed by RT-PCR. Results The biomarkers of podocyte lesion and of PT dysfunction were increased, even in normoalbuminuric type 2 DM patients. Serum and urinary IL-6 and IL-17 showed increased levels in type 2 DM patients, across all groups studied. The model provided by univariable regression analysis showed that IL-6 and IL-17 correlated directly with biomarkers of podocyte injury (nephrin, podocalyxin, synaptopodin), of PT dysfunction (KIM-1, NAG, alpha 1-microglobulin), as well as with UACR. Negative correlations have been identified regarding eGFR. In multivariable regression analysis, serum IL-6 correlated directly with synaptopodin, NAG, and negatively with eGFR (p&lt;0.00001, R2=0.805); serum IL-17 correlated directly with synaptopodin, NAG, KIM-1, UACR, and negatively with eGFR (p&lt;0.00001, R2=0.941); urinary IL-6 correlated directly with synaptopodin, NAG, and negatively with eGFR (p&lt;0.00001, R2=0.889); urinary IL-17 correlated directly with synaptopodin, nephrin, NAG, and negatively with eGFR (p&lt;0.00001, R2=0.905). Also, important associations were found between specific interleukins and miRNAs. In univariable regression analysis, IL-6 and IL-17 correlated directly with miRNA-21 and miRNA-124, and negatively with miRNA-146a and miRNA-192. The models provided by multivariable regression analysis showed that urinary IL-6 correlated directly with urinary miRNA-21, and negatively with urinary miRNA-192 (p&lt;0.00001, R2=0.886). Urinary IL-17 displayed direct correlations with urinary miRNA-21, and negative correlations with urinary miRNA-192 (p&lt;0.00001, R2=0.860). Serum IL-6 correlated directly with serum miRNA-21, miRNA-124, and indirectly with serum miRNA-146a, miRNA-192 (p&lt;0.00001, R2=0.862). Serum IL-17 showed direct correlations with serum miRNA-21, miRNA-124, and negative correlations with serum miRNA-192 (p&lt;0.00001, R2=0.745). Conclusion In the early stages of DKD, there is an association of pro-inflammatory cytokines with specific miRNAs, and with biomarkers of podocyte injury and of PT dysfunction. IL-6 and IL-17, as well as dysregulated miRNA-21, miRNA-124, miRNA-146a, and miRNA-192 display a particular molecular pattern, in relation to complex mechanisms that can initiate and maintain the chronic inflammatory response in DKD. Routine detection of these interleukins may provide biomarkers to refine the diagnosis of early renal involvement in the course of type 2 DM, independently of albuminuria and level of renal function.