Metabolomics data from liver tissue of male mice with hepatic-specific repression of the miR-379/miR-410 cluster

Description

The miR-379/miR-410 cluster, belonging to the Dlk1-Dio3 locus, is the largest cluster of miRNAs in mammals. Most of its components are predicted to regulate insulin signaling, and we found consistent upregulation of the cluster in the livers of obese subjects as well as of different experimental models of insulin resistance. The distinct miRNAs from the cluster are processed from two lncRNAs, Mirg and LncMGC. We have engineered a tool, a TuD, able to target both primary transcripts in a liver-specific manner. As a result, the cluster is repressed, which activates insulin signaling in the liver and leads to enhanced hepatic insulin sensitivity and reduced hepatic glucose output. This dataset includes metabolomics data from liver tissue of young male mice carrying hepatic-specific expression of a TuD inhibitor able to repress the miR-379/miR-410 cluster (n=8) or a negative control sequence (n=7). Samples were taken after a 6 h fasting from regular chow-fed animals.

Steps to reproduce

Fragments (≈ 20 mg) of liver samples were extracted in 100% methanol solution (190 µl) containing ribitol (0.02 mg/ml) at 70°C with vigorous shaking for 15 min. Then, 100 µl of a 100% chloroform solution was added, and the samples were shaken at 37 C for 5 extra min. Polar and organic phases were separated by centrifugation (11000 g for 10 min) after the addition of 200 µL HPLC-grade water. The polar upper phase was then transferred to a fresh vial and dried using the Eppendorf Concentrator Plus. Sequential methoximation and silylation reactions were performed using a MPS autosampler (Gerstel, Mülheim Ruhr) before loading the samples into the GC-ToF system used for GC/MS analysis. This system included an Agilent 7890 Gas Chromatograph (Agilent, Santa Clara) fitted with a Rxi-5Sil MS column (Restek Corporation, Bellefonte) coupled to a Pegasus BT Mass Spectrometer (LECO Corporation, Michigan). The GC was operated with an injection temperature of 250°C and 1 µL sample was injected in splitless mode. The ToF mass spectrometer was operated with ion source and interface temperatures of 250°C, a solvent cut time of 9 min and a scan range (m/z) of 50 – 600 with an acquisition rate of 17 spectra/second. The ChromaTof v5.50 software (LECO Corporation, Michigan) was used for data processing. The resulting signal intensities (peak area) were transformed and analyzed using Perseus software.

Institutions

• Helmholtz Zentrum München

  Bavaria, Munich

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