Summary of Study ST002403
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001532. The data can be accessed directly via it's Project DOI: 10.21228/M8P11T This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST002403 |
| Study Title | Deep multi-omic profiling reveals extensive mitochondrial remodeling driven by glycemia in early diabetic kidney disease (Mitochondria) |
| Study Summary | Changes in mitochondrial energy metabolism are thought to be central to the development of diabetic kidney disease (DKD); however, whether this response is explicitly driven by systemic glucose concentrations remains unknown. Here, we show that titrating blood glucose concentrations in vivo directly impacts mitochondrial morphology and bioenergetics and remodels the mitochondrial proteome in the kidney in early DKD. Mitoproteomic analysis revealed profound metabolic disturbances induced by severe hyperglycemia, including upregulation of enzymes involved in the TCA cycle and fatty acid metabolism, enhanced ketogenesis as well as extensive dysregulation of the mitochondrial SLC25 transporter family. The metabolite and lipid landscape were perturbed by severe hyperglycemia; untargeted metabolomics and lipidomics confirmed the enrichment of TCA cycle metabolites, an increase in triglyceride concentrations, and extensive and specific cardiolipin remodeling. Lowering blood glucose to moderate hyperglycemia stabilized all three omic landscapes, partially prevented changes in mitochondrial morphology and bioenergetics, and improved kidney injury. This study provides insights into altered substrate utilization and energy generation in the kidney early in diabetes, during moderate and severe hyperglycemia and has implications for therapeutic strategies aiming at the reinvigoration of mitochondrial function and signaling in diabetes. |
| Institute | Baker Heart and Diabetes Institute |
| Laboratory | Metabolomics |
| Last Name | Huynh |
| First Name | Kevin |
| Address | 75 Commercial Road, Melbourne, 3004 |
| kevin.huynh@baker.edu.au | |
| Phone | 0385321537 |
| Submit Date | 2022-11-20 |
| Num Groups | 3 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-12-01 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN003917 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1290 Infinity II |
| Column | Agilent ZORBAX Eclipse Plus C18 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | Agilent 6490 QQQ |
| Ion Mode | POSITIVE |
| Units | pmol per mg |
MS:
| MS ID: | MS003656 |
| Analysis ID: | AN003917 |
| Instrument Name: | Agilent 6490 QQQ |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Lipid extraction. Muscle homogenates/mitochondrial isolates were extracted using a modified single-phase chloroform/methanol extraction as described previously [Weir et al. 2016]. In brief, 20 volumes of chloroform:methanol (2:1) was added to the sample along with a series of internal standards. Samples were vortexed and spun on a rotoary mixer for 10 minutes. After sonication on a sonicator bath for 30 minutes, samples were rested for a further 20 minutes prior to centrifugation at 13,000 x g for 10 minutes. Supernatants were transferred into a 96 well plated, dried down and reconstituted in 50L water saturated butanol and sonicated for 10 minutes. After the addition of 50l of methanol with 10mM ammonium formate, the samples were spun down again at 4000RPM on a plate centrifuge (Heraeus multifuge 1S-R, ThermoFisher) and transferred into glass vials with inserts for mass spectrometry analysis. Targeted lipidomics analysis. Liquid chromatography tandem mass spectrometry (LC-MS/MS) was performed according to previously published methods, with slight modification for tissue samples [Huynh et al. 2019]. Sample extracts were analysed using either (i) a 4000 QTRAP mass spectrometer (Sciex) for cardiolipins as described preciously [Tan et al. 2020] or (ii) an Agilent 6490 QQQ mass spectrometer all other lipid species. Lipids run on the Agilent 6490 were measured using scheduled multiple reaction monitoring with the following conditions: Isolation widths for Q1 and Q3 were set to “unit” resolution (0.7 amu), gas temperature, 150°C, nebulizer 20psi, sheath gas temperature 200°C, gas flow rate 17L/min, capillary voltage 3500V and sheath gas flow 10L/min. The list of MRMs used and chromatographic conditions were extensively described previously [Huynh et al. 2019] |
| Ion Mode: | POSITIVE |
