Project description:The objective of these experiments is to identify novel direct and indirect targets of miR-150-5p in breast cancer cell lines. The goal is that these will give direction as to what targets or pathways may be contributing to the reduced growth observed in these cell lines upon restoration of miR-150-5p. A therapy directed towards one or more critical subtype-specific targets could be developed as a therapeutic for breast cancer patients. Using has-miR-150-5p mirVana miRNA mimic (Ambion, 4464066), miR-150-5p was restored to a triple negative breast cancer cell line, BT-549.
Project description:The objective of these experiments is to identify novel direct and indirect targets of miR-150-5p in breast cancer cell lines. The goal is that these will give direction as to what targets or pathways may be contributing to the reduced growth observed in these cell lines upon restoration of miR-150-5p. A therapy directed towards one or more critical subtype-specific targets could be developed as a therapeutic for breast cancer patients. Using has-miR-150-5p mirVana miRNA mimic (Ambion, 4464066), miR-150-5p was restored to an estrogen receptor positive breast cancer cell line, ZR-75-1.
Project description:MiRNAs have been shown to alter both protein expression and secretion in different cellular contexts. By combining in vitro, in vivo and in silico techniques, we demonstrated that overexpression of pre-miR-1307 reduced the ability of breast cancer cells to induce endothelial cell sprouting and angiogenesis. However, the molecular mechanism behind this and the effect of the individual mature miRNAs derived from pre-miR-1307 on protein secretion and is largely unknown. Here, we overexpressed miR-1307-3p|0, -3p|1 and 5p|0 in MDA-MB-231 breast cancer cells and assessed the impact of miRNA overexpression on protein secretion by Mass Spectrometry. Unsupervised hierarchical clustering revealed a distinct phenotype induced by overexpression of miR-1307-5p|0 compared to the controls and to the 5’isomiRs derived from the 3p-arm. Together, our results suggest different impacts of miR-1307-3p and miR-1307-5p on protein secretion which is in line with our in vitro observation that miR-1307-5p, but not the isomiRs derived from the 3p-arm reduce endothelial cell sprouting in vitro. Hence these data support the hypothesis that miR-1307-5p is at least partly responsible for impaired vasculature in tumors overexpressing pre-miR-1307.
Project description:We report the RNA-seq based analyses of the transcriptional changes in the Aedes aegypti transcriptome 4 days after blood feeding with 2 μM hsa-miR-150-5p antagomir or mock.
Project description:MicroRNAs (miRNAs) are endogenous and small non-coding RNA molecules that mediate post-transcriptional gene suppression by incomplete matches with their host mRNAs. Recent studies have highlighted that miRNAs are involved in the pathology of liver lipid metabolism, metabolic inflammation, oxidative stress, chronic liver injury, regeneration and fibrogenesis. During liver fibrosis, miRNAs are simply categorized into pro-fibrotic or anti-fibrotic miRNAs according to their expression alterations in one liver cell type especially hepatic stellate cells (HSCs). Our current study observed a miRNA, termed miR-150-5p, exhibited cell-specific expression pattern in liver fibrosis. Although the expression level of miR-150-5p in fibrotic liver tissues or hepatocytes was significantly enhanced, it was notably down-regulated in HSCs in the context of liver fibrosis. Perturbation of miR-150-5p evidently affected the cell viability of HSCs but had no obvious effect on hepatocytes. While overexpression of miR-150-5p could sensitize the apoptosis of hepatocytes in the presence of pro-apoptotic factors. Further investigations revealed that miR-150-5p mimic treatment had a larger influence on the transcriptomic stability of HSCs but a minor effect on hepatocytes. The genes related to anti-apoptosis or pro-proliferation were decreased and genes associated with pro-apoptosis or anti-proliferation were increased after miR-150-5p overexpression, which might be the potential targets of miR-150-5p overexpressed in HSCs. Collectively, our study provide a strong supporting evidence that some miRNAs may express or function in a cell-specific pattern in the backdrop of liver fibrosis.
Project description:This is a prospective-retrospective study to determine if the expression of the miRNA’s miR-31-3p and miR-31-5p are prognostic of patient outcomes or predictive of the benefit from anti-EGFR therapy in stage III Colon Cancer. The present study will utilize FFPE tumor samples collected from patients enrolled in the PETACC-8 study conducted by the Fédération Francophone de Cancérologie Digestive (FFCD). This phase 3 clinical trial prospectively randomized fully resected stage III colon cancer patients to receive adjuvant treatment with either FOLFOX-4 plus cetuximab or FLOFOX-4 alone.
Project description:Background: In patients with left heart failure, microRNAs (miRNAs) have been shown to be of diagnostic and prognostic value. The present study aims to identify miRNAs in patients with univentricular heart (UVH) disease that may be predictive of overt heart failure. Methods: A large panel of human miRNA arrays were used to determine miRNA expression profiles in the blood of 48 UVH patients and 32 age and gender-matched healthy controls. For further selection, the most abundantly expressed miRNA arrays were related to clinical measures of heart failure and selected miRNAs validated by Reverse transcription quantitative polymerase chain reaction (RT-qPCR). These validated miRNAs were used for the prediction of overt heart failure and all-cause mortality. Results: According to microarray analysis, 50 miRNAs were found to be significantly abundant in UVH patients of which miR-150-5p was best related to heart failure parameters. According to ROC analysis, NT-proBNP levels (AUC 0.940, 95% CI 0.873-1.000; p=0.001), miR-150-5p expression levels (AUC 0.905, 95% CI 0.779-1.000; p=0.001) and a higher NYHA class ≥ III (AUC 0.893, 95% CI 0.713-1.000; p=0.002) were the 3 most significant predictors of overt heart failure. Using a combined biomarker model, AUC increased to 0.980 indicating an additive value of miR-150-5p. Moreover, in the multivariate analysis, a higher NYHA class ≥ III (p=0.005) and miR-150-5p (p=0.006) turned out to be independent predictors of overt heart failure. Conclusion: In patients with UVH, miR-150-5p has additive value to natriuretic peptides for the prediction of overt heart failure and thus may be used as additional biomarker for risk assessment in these patients.
Project description:Human CD4+ Total memory cells (CD4+CD25-CD45RA-CCR7+/-) were activated for 5d with anti-CD3/anti-CD28 antibodies and transfected with biotinylated miR-150 mimic or biotinylated mimic negative control by Exiqon. After 24h cells were lysed and miRNA:mRNA complexes pulled-down with streptavidin agarose O/N at 4°C. RNA was then purified using TRI-reagent and RNA zymospin columns. Three independent donors were used for this experiment. Inputs were also sequenced. Total RNA was sequenced in single end mode. This experiment is assessing direct targets bound by miR-150-5p in human T cells.
Project description:We examined the function of miR-150 in T-cell lymphomagenesis. We transfected GFP-control or GFP-miR-150 into several T-cell lymphoma lines and sought which genes were regulated by miR-150.