miRNA Expression Research Articles (Page 1)

Ertugliflozin ameliorates experimentally-induced colitis in rats by regulating the interplay between M1/M2 macrophage polarization, tight junction proteins, and MicroRNA 155 expression.

Unveiling the immunomodulatory mechanisms of fever-range whole-body hyperthermia in humans: Insights from miRNA and protein mediators.

MicroRNAs (miRNAs) have emerged as promising biomarkers for the identification of forensically significant body fluids, aiding crime scene reconstruction and the identification of biological material donors. However, despite their potential, inter-study discrepancies and lack of reproducibility have limited their forensic application, particularly in body fluid identification. A major challenge lies in the variability introduced by different nucleic acid extraction methods. This study aimed to evaluate the co-extraction performance and miRNA detection efficiency of three commonly used nucleic acid extraction kits (AccuPrep Genomic DNA Extraction Kit, QIAamp DNA Mini Kit, and miRNeasy Tissue/Cells Advanced Micro Kit) across five saliva input volumes (400 µL, 200 µL, 100 µL, 50 µL, and 25 µL). To assess yield, purity, robustness (CV%), and sensitivity, two quantification platforms were used, followed by a two-step reverse transcription quantitative polymerase chain reaction (RT-qPCR) protocol to evaluate the impact of extraction methods on miRNA expression. Our results demonstrate that both the extraction procedure and the quantity of input material significantly influences nucleic acid recovery. Additionally, we observe variations in miRNA levels depending on the extraction method used. Surprisingly, the kit designed specifically for miRNA extraction yielded relatively poor miRNA recovery. In contrast, the DNA extraction kit (AccuPrep Genomic DNA Extraction Kit) produced the highest nucleic acid yield with moderate purity and showed the lowest Cq values for miRNA targets, indicating better miRNA detection. These findings underscore the importance of the choice of extraction kit, as it can significantly influence both the yield and quality of nucleic acids detected, and the extension, accuracy and reliability of miRNA-based forensic analyses.

Pseudoexfoliation syndrome (PXS) is associated with increased risk of glaucoma, but the underlying molecular mechanisms remain unclear. This study aimed to compare microRNA (miRNA) expression profiles between PXS with glaucoma (PXSG) and PXS without glaucoma (PXSWG). We enrolled 24 PXS patients undergoing cataract surgery, dividing them into PXSG (n=16) and PXSWG (n=8) groups. miRNA expression in anterior lens capsule tissue was analysed using NanoString nCounter technology. Differentially expressed miRNAs were identified, and functional pathway analysis was conducted using the Kyoto Encyclopedia of Genes and Genomes (KEGG). The correlations between miRNA expression and clinical variables, including glaucoma severity, endothelial cell counts (ECCs) and systemic factors identified in serum blood tests, were also examined. Using a panel of 827 miRNAs, 23 upregulated miRNAs in PXSG were identified, miRNA-(miR-)887-3 p and miR-933 exhibiting the highest differential expression. The KEGG highlighted enrichment in pathways related to ageing and signal transduction. Elevated levels of several miRNAs, miR-933 and miR-302a-3p, were linked to worse visual field (VF) and thinner peripapillary retinal nerve fibre layer thickness (pRNFLT). Multivariate regression analysis identified associations of miR-302a-3p with lower ECC, miR-302f with thinner pRNFLT and miR-614 with higher triglyceride levels. This study indicates potential differences in miRNA expression between PXSG and PXSWG, with several showing suggestive associations with key clinical parameters. These preliminary findings may provide valuable insights into processes relevant to PXS and glaucoma but require validation in larger, independent cohorts to clarify their biomarker potential.

Background/Objectives: Type 2 diabetes mellitus (T2DM) is a complex metabolic disorder characterized by insulin resistance, impaired insulin secretion, and chronic hyperglycemia. Recent studies have identified microRNAs (miRNAs), a class of small non-coding RNAs that regulate gene expression at the post-transcriptional level, as modulators of pathways involved in T2DM pathophysiology. Dysregulated miRNA expression has been detected in various samples collected from patients with T2DM, implicating these molecules in disease onset and progression. Methods: We systematically searched PubMed, Scopus, and Web of Science for studies published from the earliest available records to 18 August 2025 using the following Boolean search terms: “miRNA AND gliclazide”, “miRNA AND glibenclamide”, “miRNA AND gliquidone”, “miRNA AND glimepiride”, “mirRNA AND metformin”, “miRNA AND pioglitazone”, “miRNA AND rosiglitazone”, “miRNA AND sitagliptin”, “miRNA AND vildagliptin”, “miRNA AND alogliptin”, “miRNA and saxagliptin”, “miRNA AND linagliptin”, “miRNA AND liraglutide”, “miRNA and dulaglutide”, “miRNA AND semaglutide”, “miRNA AND tirzepatide”, “miRNA AND lixisenatide”, “miRNA AND empagliflozin”, “miRNA AND dapagliflozin”, miRNA AND insulin glargine”, “miRNA AND insulin detemir”, “miRNA AND insulin degludec”, “miRNA AND insulin aspart”, “miRNA AND insulin glulisine”, and “miRNA AND insulin lispro”. Additionally, gray literature was searched in ClinicalTrials.gov, the EU Clinical Trials Register (EudraCT), and the ISRCTN Registry to identify unpublished studies. Studies were eligible for inclusion if they were clinical interventional studies assessing the impact of currently available antidiabetic treatments on miRNA expression. Only articles published in English were considered. The risk of bias was evaluated using the RoB2 (Risk of Bias 2) and ROBINS-I (Risk Of Bias In Non-randomized Studies—of Interventions) tools. Study characteristics and major findings were tabulated. Results: A total of 1264 manuscripts was identified initially. After removing duplicates, 726 articles remained for further screening. Ultimately, 17 manuscripts reporting interventional clinical trials on the effects of antidiabetic treatment on miRNA were included, encompassing a total of 1093 patients. Key findings included treatment-associated changes in miRNA expression and their potential utility for the prediction of clinical outcomes. Conclusions: Current evidence supports the hypothesis that antidiabetic treatments modulate miRNA expression, with some findings showing predictive value for metabolic outcomes. However, the available data remain limited and of low grade of certainty, and further large-scale clinical studies are needed to provide deeper insights into these associations.

Background Thyroid nodules are common, and the majority are of benign nature, however the main clinical aim remains to discriminate malignant ones. Despite that, the actual routine workup for thyroid nodules including hormonal testing, ultrasound scan, scintigraphy, and Fine needle aspiration cytology (FNAC) is still deficient, often necessitating surgical intervention for a conclusive histopathological examination and diagnosis. Therefore, searching for molecular tools that help in differentiating benign and malignant thyroid nodules gains an increasing interest. Methodology This case-control study was conducted on 46 adults, categorized into two groups (23 patients each) one with benign and another with malignant thyroid nodules, based on post- operative histopathology. Prior to surgery patients have undergone clinical examination and ultrasonography (US) neck and only patients with US - TIRADs 3, 4 and 5 thyroid lesions were referred for FNAC and included in this study. Surgery decisions were made according to FNAC Bethesda classification. Total RNA was extracted from plasma samples prior to surgery. The reverse transcription of miRNA to cDNA was followed by Real Time PCR Amplification in triplet to analyze miRNA-222, miRNA-146b and reference miRNA-16. Plasma from 40 healthy subjects was used as control for the calculations of the relative expression of miRNA using RT-qPCR. Results The results showed a statistically significant increased expression of plasma miRNA-222 and miRNA-146b in the malignant nodules group than benign group (p = 0.001 and p= <0.001, respectively). Also, there is a statistically significant difference between benign and malignant groups regarding the presence of plasma miRNA-222 or plasma miRNA-146b with higher levels than Youden Index cutoffs (p= <0.001). MiRNA-222 showed increased plasma expression with vascular invasion of malignant nodules. Adding the estimation of plasma miRNA-222 and 146b levels to the actual ultrasound features and to FNAC Bethesda classification, raised AUC of ROC of the mentioned tools for the diagnosis of malignant thyroid nodules. Conclusion These findings suggest a potential role for miRNA-222 and miRNA-146b in differentiating benign and malignant thyroid nodules and may provide a valuable diagnostic addition to the current routine workup.

This preliminary study aimed to identify microRNA (miRNA) signatures associated with bipolar disorder (BD) by profiling blood-derived extracellular vesicles (EVs) of both putative neuronal origin and from all sources. In two parallel studies of individuals with BD and controls (CON), we characterized miRNA expression profiles of blood EVs selected for L1CAM, a putative marker of neuronal origin (n = 20 BD/20 CON), as well as bulk EVs (n = 21 BD/20 CON). For each study, analyses identified miRNAs differentially expressed between groups, followed by functional interrogation and testing for associations with clinical features. Results of Study 1 showed 34 miRNAs differentially expressed between groups and implicated PTEN, a gene whose protein levels were previously found to be altered in postmortem brain studies of BD. Results of Study 2 showed 10 miRNAs differentially expressed between groups. Limited overlap was identified between studies, with only hsa-miR-1-3p identified with the same direction of change across both types of EVs. Differentially expressed miRNAs were significantly associated with clinical features of BD only in Study 1. Our results, albeit preliminary, reiterate a crucial role for miRNAs in the pathophysiology of BD and suggest that miRNA signatures of putative neuronal origin may more closely correspond to clinical features.

Background: Adipose tissue-derived extracellular vesicles (adiposomes) are key mediators of intercellular communication and play a critical role in metabolic disease progression, including diabetes. These vesicles carry bioactive molecules such as microRNAs (miRNAs) that regulate inflammation, vascular function, and metabolism. Our previous work showed that adiposomes from obese diabetic individuals impair endothelial function, promote vascular remodeling, and reduce microvascular flow-induced dilation, contributing to cardiovascular disease (CVD). However, the adiposomal cargo underlying these effects remains unknown. This study examines adiposomal miRNA profiles as potential contributors to the elevated CVD risk in obese diabetic patients. Methods: Adipose tissue samples were collected from obese individuals and classified into non-diabetic (n=17), pre-diabetic (n=10), and diabetic (n=14) groups based on fasting glucose and hemoglobin A1C. Adiposomes were isolated and characterized using nanoparticle tracking analysis and electron microscopy. miRNAs were extracted, sequenced, and analyzed bioinformatically. Correlations were assessed between miRNA expression and cardiometabolic measures, including (1) body fat and lean mass (DEXA), (2) glucose and lipid profiles, (3) brachial artery flow-mediated dilation, (4) flow-induced dilation in isolated arterioles, (5) echocardiographic cardiac function, and (6) inflammatory and endothelial biomarkers (IL-6, CRP, nitric oxide). Results: Diabetic and pre-diabetic groups exhibited distinct adiposomal miRNA profiles compared to non-diabetic controls. Top differentially expressed miRNAs included miR-361, miR-21, miR-130a, miR-181a, miR-10a, miR-126, miR-30e, miR-92b, miR-125a, and miR-221. Many of these miRNAs are implicated in hypertension, atherosclerosis, and cardiac hypertrophy. miR-221, miR-361, and miR-21 were upregulated 3–4 fold in diabetic subjects and significantly correlated with HbA1C and HOMA-IR. Regression analyses revealed associations between these miRNAs and BMI, fat percentage, impaired vascular dilation, and elevated LDL. Conclusion: Dysregulated adiposomal miRNA cargo may contribute to cardiometabolic dysfunction in obesity-associated diabetes. These findings highlight adiposomal miRNAs as potential mechanistic links and biomarkers for diabetes-related cardiovascular risk. Ongoing studies are investigating the downstream pathways regulated by these miRNAs.

Introduction Circulating microRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and play key roles in metabolic diseases, including diabetes. This study aimed to identify circulating miRNA signatures linked to glycemic status, vitamin D deficiency, aspirin consumption, and platelet activity in individuals with type 2 diabetes, prediabetes, and healthy controls. Methods Plasma samples from 24 participants (14 with diabetes, 2 prediabetic, and 8 non-diabetic controls) were analyzed using next-generation sequencing to assess differences in miRNA expression. Results Although principal component analysis showed no group separation, pairwise testing identified 131 miRNAs that were significantly altered between the diabetic and control groups, as well as 141 miRNAs between the prediabetic and control groups (FDR &amp;lt;0.05), with 56 overlapping between contrasts. Fourteen miRNAs in the pre-diabetic group and twelve in the diabetic group overlapped with literature-supported candidates, reinforcing their clinical relevance. In the direct diabetes vs. prediabetes comparison, no miRNAs passed FDR. Still, consistent trend-level decreases in diabetes were observed for hsa-miR-4776-5p, hsa-miR-6778-3p, and hsa-miR-5002-3p, which were among the most upregulated in prediabetes vs. controls. Sex-stratified analyses revealed distinct miRNA expression patterns, emphasizing the influence of biological sex on miRNA regulation in glucose metabolism. Correlation analyses highlighted miRNAs associated with glucose/HbA1c, alongside relationships among vitamin D status, glycemia, and platelet function. Discussion Collectively, these data highlight the exploratory potential of circulating miRNAs as candidate early biomarkers for glycemic dysregulation, providing a foundation for future validation and development of improved diagnostic and preventive strategies for type 2 diabetes.

Introduction Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer-related deaths worldwide, largely due to complex interactions within the tumor-immune microenvironment that limit treatment efficacy. MicroRNAs (miRNAs) play critical roles in the regulation of tumor progression and immune evasion. This study systematically evaluated the expression characteristics, clinical significance, and role of miR-4664-3p in tumor immune regulation in NSCLC. Methods We analyzed an NSCLC dataset from The Cancer Genome Atlas (TCGA) and identified miR-4664-3p as a potential diagnostic, prognostic, and immunotherapeutic biomarker. Bioinformatic approaches have been used to assess miRNA expression and clinical significance. The regulatory role of the miR-4664-3p/Protein Kinase C Beta (PRKCB) axis was further examined using correlation analysis, nomogram construction, and experimental validation in cell lines and animal models. Results MiR-4664-3p was significantly upregulated in NSCLC tissues and served as an independent predictor of poor prognosis. Its increased expression was linked to reduced immune cell infiltration and enhanced immune escape. PRKCB was validated as a direct downstream target of miR-4664-3p and showed a positive association with CD8 + T cell infiltration and favorable outcomes. Functional assays confirmed that miR-4664-3p promoted NSCLC cell proliferation, migration, and invasion. Conversely, the inhibition of miR-4664-3p increased PRKCB expression, boosted CD8 + T cell activity, strengthened anti-tumor immunity, and suppressed tumor growth. Conclusion These results suggest that the miR-4664-3p/PRKCB axis is crucial in NSCLC progression and immune modulation. Hence, MiR-4664-3p is a potential diagnostic and prognostic indicator, as well as therapeutic target in immunotherapy strategies for NSCLC.

Nonalcoholic steatohepatitis (NASH) pathogenesis involves apoptosis, inflammation, lipogenesis, and insulin resistance. Hepatocyte apoptosis leads to liver injury and fibrosis, while inflammation exacerbates damage through immune cells and cytokines. Lipogenesis promotes fat accumulation and lipotoxicity, stressing hepatocytes, whereas insulin resistance impairs glucose metabolism and exacerbates dysfunction. Highlight the complexity of NASH and the need for multi-targeted therapies are highlighted by these interrelated processes. Using in silico transcriptomic analysis, this study identifies key microRNAs (miRNAs) in NASH and explores the relationship between miRNAs and circ-MYO9B in NASH. To find differentially expressed miRNAs (DEMs), miRNA expression profiles from the liver and blood datasets (GSE33857, GSE59492, and GSE49012) were examined. Nonalcoholic steatohepatitis-induced cirrhosis (NASH-CH)-related miRNA modules were discovered by Weighted Gene Co-expression Network Analysis (WGCNA). RT-qPCR was used on tissue and blood samples to examine the expression and interaction of circ-MYO9B and hsa-miR-193b-5p in NASH patients, and dual-luciferase assays confirmed their direct interaction. Circ-MYO9B and hsa-miR-193b-5p were found to be significantly correlated in both blood and liver tissue in NASH-CH patients, according to expression analysis. RT-qPCR analysis validated their dysregulation, suggesting their role in the advancement of the disease. The direct interaction between circ-MYO9B and hsa-miR-193b-5p was confirmed by the dual-luciferase assay, indicating a regulatory function in metabolic pathways. By modulating this miRNA, circ-MYO9B may influence downstream pathways in which miR-193b-5p is known to be involved, such as lipogenesis, inflammation, apoptosis, and glucose metabolism. These findings suggest that circ-MYO9B may represent a potential therapeutic target for addressing metabolic dysfunction associated with NASH, although further validation in hepatocyte-specific models is warranted.

ABSTRACTGitelman syndrome (GS) is caused by an inactivating mutation in the SLC12A3, encoding the thiazide‐sensitive sodium chloride cotransporter (NCC), leading to salt wasting and electrolyte imbalance. Our aim is to identify microRNA (miRNA) expression and explore their role from urinary extracellular vesicles (uEVs) in GS patients. In this study, both uEVs from 23 genetically confirmed GS patients and renal biopsied tissues from another 3 GS patients were extracted for small RNA sequencing. Small RNA sequencing identified 358 miRNAs from uEVs and 652 miRNAs from renal biopsied tissues. Among differentially expressed miRNAs, 20 were upregulated and 23 downregulated in uEVs, while renal biopsied tissues showed 30 upregulated and 23 downregulated miRNAs. Four miRNAs (hsa‐let‐7d‐3p, hsa‐miR‐362‐5p, hsa‐miR‐30c‐5p, and hsa‐miR‐30b‐5p) overlapped from uEVs and renal tissues. In particular, the terms of the distinct upregulated hsa‐let‐7d‐3p target genes were related to ion transport and membrane depolarization, especially in neural precursor cell expressed, developmentally downregulated 4‐like (NEDD4L). Real‐time PCR of uEVs from another 11 GS patients confirmed significantly elevated hsa‐let‐7d‐3p compared to healthy controls. The decrease in the Nedd4l expression in the collecting duct was also confirmed in NccS707X/S707X knock‐in mice. Dual luciferase assays further demonstrated that hsa‐let‐7d‐3p negatively regulated the expression of NEDD4L. These findings concluded that differentially expressed miRNAs could be identified from uEVs in GS patients, and hsa‐let‐7d‐3p, the only upregulated miRNA, negatively regulates NEDD4L expression in the collecting duct.

Cardiovascular disease (CVD) causes more than 50% of deaths in patients with advanced chronic kidney disease (CKD). Clinical studies suggest that kidney-derived factors contribute to CVD development in CKD, independently of co-morbidities. However, to date, no kidney-specific humoral risk factor that triggers direct cardiotoxicity has been identified. In this cross-sectional study, we investigate how, in CKD patients, circulating extracellular vesicles (EVs) facilitate pathological kidney-heart communication, thereby causing cardiotoxicity, impairing cardiac function, and contributing to heart failure (HF) progression. We investigated the function of EVs from CKD patients and adenine diet-induced CKD mice on cardiomyocyte and cardiac contractility. MiRNA cargo of EVs was identified by small RNA sequencing and qRT-PCR, and their cardiotoxicity was tested by using miRNA-mimics. Tissue and cellular origin of CKD-EV-miRNAs were determined from their corresponding primary miRNA expressions in mice. EVs from plasma of CKD patients, but not from healthy controls, were cardiotoxic; they significantly induced apoptosis both in vitro and in vivo and impaired contractility of adult rat primary cardiomyocytes in vitro. Likewise, EVs from both plasma and kidneys of CKD mice were cardiotoxic. Pharmacologically depleting circulating EVs in CKD mice significantly recovered cardiac function and ameliorated HF, improvements that suggest CKD-EVs play a causal role in HF pathogenesis. Both human and mouse CKD-EVs were enriched in distinct miRNAs, compared to control-EVs. CKD-EV-miRNA mimics were cardiotoxic, impairing contractility and downregulating contractile gene expression in human-iPSC-derived cardiomyocytes. Interestingly, levels of endogenous primary miRNAs corresponding to circulating CKD-EV-miRNAs were significantly higher in CKD-kidney tissues, specifically in CD45-veCD31-ve renal cells, but not in CKD-hearts, CKD-livers or CKD-PBMCs from peripheral blood, a result that indicates that CKD-EV-miRNAs originate renally. Remarkably, CKD-EV-miRNA levels correlated with established markers of cardiac injury, thus uncovering the presence of sub-clinical heart disease and demonstrating heterogeneity in reno-cardiac disease. Collectively, our human subject and mouse studies show that circulating CKD-EVs, carrying distinct renal-derived miRNAs, mediate the molecular crosstalk that contributes to the pathogenesis of HF in CKD. Consequently, CKD-EVs hold promise as diagnostic and prognostic biomarkers for early disease detection and as targets for novel therapeutic interventions in chronic reno-cardiac disease.

In this study, we aimed to validate the potential of miR-374 in ameliorating hyperglycemia by regulating peroxisome proliferator-activated receptor γ coactivator-1 (PGC-1α) expression in pancreatic islets and liver. To identify miRNAs targeting PGC-1α, we performed miRNA chip analysis in rat islets under hyperglycemic and euglycemic conditions. Luciferase reporter assay was performed to identify miR binding sites in the 3'-untranslated region (3' UTR) of PGC-1α. In db/db mice, miRNA-encapsulated adenoviruses were administered and intraperitoneal glucose tolerance test and glucose stimulated insulin secretion tests were performed. For enhanced delivery to β-cells, we developed exendin-4 (Ex-4) coated cationic lipoparticles (CCLs) encapsulating miRNAs. The therapeutic potential of Ex-4-CCL-miRNA was further evaluated in insulin-producing cells derived from induced pluripotent stem cells. By analyzing miRNA expression in primary rat islets exposed under hyperglycemic environment, we identified miR-374 as a potential target. In vitro experiments confirmed that miR-374 significantly suppressed PGC-1α expression in β-cells and hepatocytes by binding to its 3'-UTR. In vivo experiments using adenovirus-mediated miR-374 (Ad-miR-374) delivering directly to the pancreas and liver of db/db mice demonstrated improved glycemic control, enhanced insulin secretion, and downregulated hepatic gluconeogenesis-related genes (G6Pase, Pepck, PC). To enhance the clinical applicability of miR-374, we developed Ex-4-CCLs. Ex-4-CCL-miR-374 successfully alleviated hyperglycemia, restored pancreatic islet function, and decreased gluconeogenesis gene expression in db/db mice. Furthermore, Ex-4-CCL-miR-374 improved insulin secretory function in glucotoxicity-exposed human induced pluripotent stem cell-derived insulin producing cells. Based on these findings, we propose that Ex-4-CCL-miR-374 as a promising therapeutic approach to reverse β-cell dysfunction and improve hepatic insulin resistance in type 2 diabetes mellitus.

Ginger is rich in various phytochemicals known for their potential antifibrotic properties, largely attributed to their anti-inflammatory effects. Although the anti-inflammatory benefits of ginger are well established, the underlying molecular mechanisms, particularly those involving microRNAs (miRNAs) at the post-transcriptional level, remain inadequately understood. This study investigated the antifibrotic effects of ginger extract in mice by examining its influence on the modulation of miR-21-5p and miR-29b, along with their respective target genes. The BALB/c mice were allocated into two separate groups at random, with each group consisting of six mice: one serving as the control group and the other as the treatment group. Both groups received standard chow and water; the treatment group was additionally given ginger extract via oral gavage, while the control group received water. Throughout the treatment period, two mice from each group succumbed. The remaining animals were subsequently utilized for downstream analyses. After three weeks, blood samples were collected for miRNA expression analysis. TargetScan was utilized to identify the target genes of both miRNAs, and their expression levels were confirmed through real time quantitative PCR (RT-qPCR). Smad7 and Col1a1 were confirmed as target genes of miR-21-5p and miR-29b, respectively. Smad7 is recognized for its antifibrotic role, while Col1a1 is a key marker associated with extracellular matrix formation. RT-qPCR results showed that ginger treatment led to the upregulation of miR‑21‑5p accompanied by increased Smad7 expression, while miR‑29b levels were also elevated, suggesting activation of antifibrotic regulatory pathways. Although changes in Col1a1 expression were less pronounced, the observed modulation of key antifibrotic miRNAs highlights ginger’s potential as a natural modulator of fibrosis‑related pathways and supports further investigation of its therapeutic benefits in fibrotic disorders.

Effects of residual PAH exposure from firefighters' skin and turnout gear on biospecimen microRNA expression.

Identification of dysregulated miRNA transcriptomic profile in canine hepatocellular carcinoma and comparison to human HCC.

Pentagalloyl glucose reverses chemoresistance in triple-negative breast cancer via EMT inhibition and miRNA modulation.

Preeclampsia is amultisystem disorder involving in inflammatory responses and metabolic dysfunction of maternal-fetal circulation. Recently, researchers found it threatens renal health of offspring in adulthood. Growing evidence indicated chronic kidney disease is associated with glomeruli deficiencies during intrauterine development. Our previous study showed placenta-derived exosomes from cord plasma with preeclampsia impede fetal glomerular vascularization, during which we postulate microRNAs may function as epigenetic switches for gene silencing of human glomerular endothelial cells. However, the specific miRNAs in placenta-derived exosomes engaged in glomerular vascularization remain unclear. Small RNA sequencing of placental-derived exosomes and bioinformatics analysis were applied to identify differentially expressed miRNAs, followed by real-time polymerase chain reaction for verification. Transient expression and inhibition of candidate miRNA were performed by transfection with chemically synthesized miRNA oligonucleotides. Functional assays of HGECs including cell proliferation assays, EDU assays, migration assays, tube formation assays and monolayer cell barrier permeability assays were performed after transfection. Further, dual luciferase assay was used to explore the target genes of candidate miRNA, followed by RT-qPCR, western blot and rescue assays. Antagomirs transfection of C57BL/6J fetal mice via Amniotic cavity injection in vivo and C57BL/6J fetal mice kidney explants culture in vitro were performed to evaluate number of glomeruli, renal development. Preeclampsia downregulates miR-199a-3p in placenta-derived exosomes from cord plasma. Suppression of endogenous miR-199a-3p in HGECs inhibits angiogenesis, proliferation, migration and permeability. A dual luciferase assay and rescue assays confirmed that miR-199a-3p targets PH domain leucine-rich repeat-containing protein phosphatase 2, regulating the phosphorylation of Akt serine/threonine kinase 1 (S473). C57BL/6J fetal mice with miR-199a-3p downregulation have low glomerulus counts and relative growth rate. miR-199a-3p in placenta-derived exosomes from cord plasma controls VEGF-induced glomerular angiogenesis. Moreover, it provides a distinct perspective for the mechanisms underlying the increased risk for renal disease in the offspring of preeclampsia patients via placenta-derived exosomal miRNAs.

Recent progress on biosensors for detection of circulating miRNA biomarkers.