miR-129-5p as a biomarker for pathology and cognitive decline in Alzheimer's disease.

BACKGROUND: Alzheimer's dementia (AD) pathogenesis involves complex mechanisms, including microRNA (miRNA) dysregulation. Integrative network and machine learning analysis of miRNA can provide insights into AD pathology and prognostic/diagnostic biomarkers. METHODS: We performed co-expression network analysis to identify network modules associated with AD, its neuropathology markers, and cognition using brain tissue miRNA profiles from the Religious Orders Study and Rush Memory and Aging Project (ROS/MAP) (N = 702) as a discovery dataset. We performed association analysis of hub miRNAs with AD, its neuropathology markers, and cognition. After selecting target genes of the hub miRNAs, we performed association analysis of the hub miRNAs with their target genes and then performed pathway-based enrichment analysis. For replication, we performed a consensus miRNA co-expression network analysis using the ROS/MAP dataset and an independent dataset (N = 16) from the Gene Expression Omnibus (GEO). Furthermore, we performed a machine learning approach to assess the performance of hub miRNAs for AD classification. RESULTS: Network analysis identified a glucose metabolism pathway-enriched module (M3) as significantly associated with AD and cognition. Five hub miRNAs (miR-129-5p, miR-433, miR-1260, miR-200a, and miR-221) of M3 had significant associations with AD clinical and/or pathologic traits, with miR129-5p by far the strongest across all phenotypes. Gene-set enrichment analysis of target genes associated with their corresponding hub miRNAs identified significantly enriched biological pathways including ErbB, AMPK, MAPK, and mTOR signaling pathways. Consensus network analysis identified two AD-associated consensus network modules and two hub miRNAs (miR-129-5p and miR-221). Machine learning analysis showed that the AD classification performance (area under the curve (AUC) = 0.807) of age, sex, and APOE ε4 carrier status was significantly improved by 6.3% with inclusion of five AD-associated hub miRNAs. CONCLUSIONS: Integrative network and machine learning analysis identified miRNA signatures, especially miR-129-5p, as associated with AD, its neuropathology markers, and cognition, enhancing our understanding of AD pathogenesis and leading to better performance of AD classification as potential diagnostic/prognostic biomarkers.

miR-129-5p as a biomarker for pathology and cognitive decline in Alzheimer's disease.

Background: Alzheimer's dementia (AD) pathogenesis involves complex mechanisms, including microRNA (miRNA) dysregulation. Integrative network and machine learning analysis of miRNA can provide insights into AD pathology and prognostic/diagnostic biomarkers. Methods: We performed co-expression network analysis to identify network modules associated with AD, its neuropathology markers, and cognition using brain tissue miRNA profiles from the Religious Orders Study and Rush Memory and Aging Project (ROS/MAP) (N = 702) as a discovery dataset. We performed association analysis of hub miRNAs with AD, its neuropathology markers, and cognition. After selecting target genes of the hub miRNAs, we performed association analysis of the hub miRNAs with their target genes and then performed pathway-based enrichment analysis. For replication, we performed a consensus miRNA co-expression network analysis using the ROS/MAP dataset and an independent dataset (N = 16) from the Gene Expression Omnibus (GEO). Furthermore, we performed a machine learning approach to assess the performance of hub miRNAs for AD classification. Results: Network analysis identified a glucose metabolism pathway-enriched module (M3) as significantly associated with AD and cognition. Five hub miRNAs (miR-129-5p, miR-433, miR-1260, miR-200a, and miR-221) of M3 had significant associations with AD clinical and/or pathologic traits, with miR129-5p by far the strongest across all phenotypes. Gene-set enrichment analysis of target genes associated with their corresponding hub miRNAs identified significantly enriched biological pathways including ErbB, AMPK, MAPK, and mTOR signaling pathways. Consensus network analysis identified two AD-associated consensus network modules, and two hub miRNAs (miR-129-5p and miR-221). Machine learning analysis showed that the AD classification performance (area under the curve (AUC) = 0.807) of age, sex, and apoE ε4 carrier status was significantly improved by 6.3% with inclusion of five AD-associated hub miRNAs. Conclusions: Integrative network and machine learning analysis identified miRNA signatures, especially miR-129-5p, as associated with AD, its neuropathology markers, and cognition, enhancing our understanding of AD pathogenesis and leading to better performance of AD classification as potential diagnostic/prognostic biomarkers.

Inhibition of MEG3 ameliorates cardiomyocyte apoptosis and autophagy by regulating the expression of miRNA-129-5p in a mouse model of heart failure.

The long non-coding RNA, maternally expressed gene 3 (MEG3), are involved in myocardial fibrosis and compensatory hypertrophy, but its role on cardiomyocyte apoptosis and autophagy in heart failure (HF) remains unclear. The aim of this study was to investigate the effect of MEG3 on cardiomyocyte apoptosis and autophagy and the underlying mechanism. A mouse model of HF was established by subcutaneous injection of isoproterenol (ISO) for 14 days, and an in vitro oxidative stress injury model was replicated with H2O2 for 6 h. SiRNA-MEG3 was administered in mice and in vitro cardiomyocytes to knock down MEG3 expression. Our results showed that cardiac silencing of MEG3 can significantly ameliorate ISO-induced cardiac dysfunction, hypertrophy, oxidative stress, apoptosis, excessive autophagy and fibrosis induced by ISO. In addition, inhibition of MEG3 attenuated H2O2-induced cardiomyocyte oxidative stress, apoptosis and autophagy in vitro. Downregulation of MEG3 significantly inhibited excessive cardiomyocyte apoptosis and autophagy induced by ISO and H2O2 through miRNA-129-5p/ATG14/Akt signaling pathways, and reduced H2O2-induced cardiomyocyte apoptosis by inhibiting autophagy. In conclusion, inhibition of MEG3 ameliorates the maladaptive cardiac remodeling induced by ISO, probably by targeting the miRNA-129-5p/ATG14/Akt signaling pathway and may provide a tool for pharmaceutical intervention.

Exosomes derived from microRNA-129-5p-modified tumor cells selectively enhanced suppressive effect in malignant behaviors of homologous colon cancer cells.

Exosome-encapsulated microRNAs (miRNAs) are novel diagnostic and predictive markers in colon cancer. Hence, the study of serum exosomal miRNAs in patients with colon cancer may help its diagnosis and treatment. PKH26-labeled exosomal uptake analysis identified whether exosomes transfer miRNA-129-5p to target cells. Transmission electron microscopy and dynamic light scattering analysis were applied to determine exosome morphology and size distribution. The Cell Counting Kit-8, wound healing assay and Transwell assays were used to detect cell proliferation, migration, and invasion after treatment with engineered exosomes. Moreover, the Western blotting was used to quantify the expression of proteins involved in cell apoptosis. In our study, hepatocellular liver carcinoma, cervical cancer and colon cancer cells were selected as the target cells of miRNA-129-5p exosomes. Exosomes containing miRNA-129-5p were found to be significantly more easily absorbed by colon cancer cells, presenting a stronger inhibitory effect on colon cancer cell proliferation. MiRNA-129-5p exosomes induced apoptosis in colon cancer cells while inhibiting their proliferation, migration, and invasion. In conclusion, exosomes derived from miRNA-129-5p-modified tumor cells selectively inhibited colon cancer progression, shedding new insights to therapeutic efficacy of this cancer.

Study on the mechanism of miRNA-129-5p in epithelial-mesenchymal transition of biliary atresia mouse / 重庆医学

Objective To investigate the mechanism of miRNA-129-5p in epithelial-mesenchymal transition (EMT)of biliary atresia.Methods Constructed bile duct epithelial EMT cell model (the experimental group) induced by TGF-β1,detected the expressions of EMT related markers and miRNA-129-5p.While miRNA-129-5p precursor was transfected,the expressions of EMT related markers and extracellular matrix were contrasted between the original and the renovated biliary epithelial cells.Results In the experimental group,extrahepatic bile duct showedEMT,the expression of miRNA-129-5p was decreased (P＜0.05),overexpression of miRNA-129-5p could inhibit the progression of EMT (P＜0.05).Conclusion miRNA-129-5p may relate to EMT by regulating the expression of TGF-β1.

MicroRNA-129-5p inhibits vascular smooth muscle cell proliferation by targeting Wnt5a.

Aberrant smooth muscle cells (SMCs) play important roles in the formation of abdominal aortic aneurysm (AAA). Although the molecular mechanism of AAA formation has been investigated, there is a lack of understanding concerning the role of microRNAs (miRNAs) in AAA, which the current study aimed to address. Firstly, miRNA array analysis was performed in order to compare the miRNA profiles in a mouse model of AAA with those in normal control mice, and differentially expressed miRNAs were identified. miR-129-5p was selected for further analysis, and was used to transfect human SMCs. The results of an MTT assay revealed that miR-129-5p inhibited the proliferation of SMCs, and flow cytometry indicated that apoptosis was induced. Bioinformatic analysis predicted that Wnt5a was the potential target gene of miR-129-5p, and this was verified by luciferase assay. In summary, miR-129-5p inhibits cellular proliferation, induces apoptosis and modulates the Wnt5a signaling pathway in SMCs.

Upregulation of miR-129-5p affects laryngeal cancer cell proliferation, invasiveness, and migration by affecting STAT3 expression.

microRNA-129-5p may have a relationship with laryngeal squamous cell carcinoma (LSCC), but the expression and function of miRNA-129-5p are not fully understood. This study was performed to investigate the expression and function of miRNA-129-5p in LSCC and its effects on STAT3. HBE and Hep-2 cells were cultured and analyzed by western blotting and real-time PCR. miR-129-5p expression was detected by real-time PCR. Hep-2 cells were transfected with a miR-129-5p expression vector. Cell proliferation, invasion, migration, cell cycle, and apoptosis assays were used to determine the role of miRNA-129-5p in the growth, invasion, and migration of LSCC cells and to determine the importance of STAT3 in these effects. STAT3 mRNA expression in Hep-2 cells was significantly higher than that in HBE cells (P < 0.05). miR-129-5p expression detected by real-time PCR showed that it was expressed at a lower level in Hep-2 cells than in HBE cells (P < 0.05). Compared with the control cells, the transfected cells showed lower STAT3 mRNA expression. For up to 7 days in culture, the transfected cells showed lower proliferation than the control cells (P < 0.05). After 24 h in culture, the apoptosis rate in miR-129-5p-transfected cells was 3.48 ± 0.38 %, while the rate in control cells was 0.92 ± 0.09 % (P = 0.0028), but the statistical significance was lost after 72 h in culture (P = 0.3180). The invasion and migration of the cells were inhibited after 24 and 72 h in culture when the miR-129-5p expression in Hep-2 cells was upregulated (P = 0.0037 and 0.00383, respectively), while there was no statistically significant difference at 48 h (P = 0.0712). STAT3 expression could be suppressed by the upregulation of miR-129-5p expression. Both the proliferation and migration of tumor cells were suppressed when the level of STAT3 expression was decreased. The apoptosis rate of tumor cells was also increased. Based on these data, we suggest that miR-129-5p may directly inhibit STAT3 expression and play an important role in the development of LSCC.

miRNA-129-5p regulates the targets of the VCP gene expression in human osteosarcoma cells / 医学研究生学报

Objective High expression of the valosin-containing protein ( VCP) gene can enhance the metastasis of osteosar-coma via the AKT/PI3K/NF-KappaB/MMP-9 signaling pathway, but the molecular mechanisms underlying the up-regulation of VCP in osteosarcoma cells remains unknown .This study aimed to determine whether miRNA-129-5p can regulate the VCP expression and its targets in human osteosarcoma cells . Methods The microRNA target-predicting software TargetScanhuman 6.2 ( http://www.tar-getscan.org/) was used to predict the possible targets of miRNA-129-5p on the VCP gene.Then, two recombinant gene report vectors containing the wild VCP gene 3′UTR ( psi-VCP vector ) and mutant VCP gene 3′UTR ( psi-VCPmut vector ) were constructed , se-quenced, and identified.The human osteosarcoma U2-OS cells were co-transfected with miRNA-129-5p mimic and psi-VCP vector or psi-VCPmut vector, respectively.A non-specificity mimic transfection served as negative control , and the luciferase activity was detec-ted in each group. Results The software prediction showed only one conserved function site of miRNA-129-5p on the VCP gene 3′UTR163-169 bp.Luciferase activity was significantly lower in the psi-VCP vector +miRNA-129-5p transfection group (15.529 ± 1.902) than in the VCP control group (21.781 ±0.854), VCP mutation experimental group (19.978 ±1.377), and VCP mutation control group (21.952 ±1.516) (P<0.05), with no remarkable difference between the VCP mutation control and VCP control groups (P=0.276). Conclusion miRNA-129-5p can probably regulate the targets of the VCP gene in human osteosarcoma U 2-OS cells.