Summary of Study ST003248

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 PR002016. The data can be accessed directly via it's Project DOI: 10.21228/M81C00 This work is supported by NIH grant, U2C- DK119886.

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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.

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Study IDST003248
Study TitleEffects of acute mitoregulin loss on cardiac mitochondrial lipids in mice
Study SummaryCardiac mitochondrial lipidome analysis in 7-week-old male aMHC-Cas9 transgenic mice, harvested at 4 weeks post-injection with either AAV:guMtln (sgRNAs to delete Mtln) or AAV:GFP control.
Institute
University of Iowa
Last NameBoudreau
First NameRyan
Address4334 PBDB, 169 Newton Rd, Iowa City, IA 52242
Emailryan-boudreau@uiowa.edu
Phone3193535573
Submit Date2024-06-05
Raw Data AvailableYes
Raw Data File Type(s)mzML
Analysis Type DetailLC-MS
Release Date2024-12-31
Release Version1
Ryan Boudreau Ryan Boudreau
https://dx.doi.org/10.21228/M81C00
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002016
Project DOI:doi: 10.21228/M81C00
Project Title:Effects of mitoregulin loss on cardiac and mitochondrial lipids in mice
Project Summary:We and others discovered a highly-conserved mitochondrial transmembrane microprotein, named Mitoregulin (Mtln), that supports lipid metabolism. We reported that Mtln strongly binds cardiolipin (CL), increases mitochondrial respiration and Ca2+ retention capacities, and reduces reactive oxygen species (ROS). Here we extend our observation of Mtln-CL binding and examine Mtln influence on cristae structure and mitochondrial membrane integrity during stress. We demonstrate that mitochondria from constitutive- and inducible Mtln-knockout (KO) mice are susceptible to membrane freeze-damage and that this can be rescued by acute Mtln re-expression. In mitochondrial-simulated lipid monolayers, we show that synthetic Mtln decreases lipid packing and monolayer elasticity. Lipidomics revealed that Mtln-KO heart tissues show broad decreases in 22:6-containing lipids and increased cardiolipin damage/remodeling. Lastly, we demonstrate that Mtln-KO mice suffer worse myocardial ischemia-reperfusion injury, hinting at a translationally-relevant role for Mtln in cardioprotection. Our work supports a model in which Mtln binds cardiolipin and stabilizes mitochondrial membranes to broadly influence diverse mitochondrial functions, including lipid metabolism, while also protecting against stress.
Institute:University of Iowa
Last Name:Boudreau
First Name:Ryan
Address:4334 PBDB, 169 Newton Rd, Iowa City, IA 52242
Email:ryan-boudreau@uiowa.edu
Phone:3193535573

Subject:

Subject ID:SU003367
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Species Group:Mammals

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Genotype Sample source
SA3534733058-1acute ko cardiac mitochondria
SA3534743058-2acute ko cardiac mitochondria
SA3534753097-3acute ko cardiac mitochondria
SA3534763097-4acute ko cardiac mitochondria
SA3534773109-1acute ko cardiac mitochondria
SA3534783109-3acute ko cardiac mitochondria
SA3534793058-3cntrl cardiac mitochondria
SA3534803058-4cntrl cardiac mitochondria
SA3534813097-1cntrl cardiac mitochondria
SA3534823109-2cntrl cardiac mitochondria
SA3534833116-1cntrl cardiac mitochondria
SA3534843116-2cntrl cardiac mitochondria
Showing results 1 to 12 of 12

Collection:

Collection ID:CO003360
Collection Summary:Mitochondria were isolated from hearts of 6 AAV9:GFP injected and 6 AAV9:guMtln injected aMHC-Cas9 transgenic mice at approximately 7 weeks of age (4 weeks post AAV injection) and mitochondrial pellets stored at -80. Protein content of mitochondrial pellets was determined by BCA assay of distinct, paired pellets (collected from the same homogenate), after detergent solubilization.
Sample Type:Heart

Treatment:

Treatment ID:TR003376
Treatment Summary:No treatment.

Sample Preparation:

Sampleprep ID:SP003374
Sampleprep Summary:Mitochondria were isolated from hearts of 6 AAV9:GFP injected and 6 AAV9:guMtln injected aMHC-Cas9 transgenic mice at approximately 7 weeks of age (4 weeks post AAV injection) and mitochondrial pellets stored at -80. Protein content of mitochondrial pellets was determined by BCA assay of distinct, paired pellets (collected from the same homogenate), after detergent solubilization.

Combined analysis:

Analysis ID AN005320 AN005321
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Agilent 1290 Infinity Agilent 1290 Infinity
Column Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
MS Type ESI ESI
MS instrument type QTOF QTOF
MS instrument name Agilent 6545 QTOF Agilent 6545 QTOF
Ion Mode POSITIVE NEGATIVE
Units pmol lipid per mg tissue pmol lipid per mg tissue

Chromatography:

Chromatography ID:CH004024
Chromatography Summary:Positive Mode RP LCMS
Instrument Name:Agilent 1290 Infinity
Column Name:Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
Column Temperature:65
Flow Gradient:Started at 15% mobile phase B then increased to 30% B over 2.4 min. It sequentially increased to 48% B from 2.4 – 3.0 min, 82% B from 3 – 13.2 min, and 99% B from 13.2 – 13.8 min where it’s held until 16.7 min and returned to the initial conditions and equilibrated for 5 min.
Flow Rate:0.4 mL min
Solvent A:40% water/60% acetonitrile; 0.1% formic acid; 10 mM ammonium formate
Solvent B:90% isopropanol/9% acetonitrile/1% water; 0.1% formic acid; 10 mM ammonium formate
Chromatography Type:Reversed phase
  
Chromatography ID:CH004025
Chromatography Summary:Negative Mode RP LCMS
Instrument Name:Agilent 1290 Infinity
Column Name:Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
Column Temperature:65
Flow Gradient:Started at 15% mobile phase B then increased to 30% B over 2.4 min. It sequentially increased to 48% B from 2.4 – 3.0 min, 82% B from 3 – 13.2 min, and 99% B from 13.2 – 13.8 min where it’s held until 16.7 min and returned to the initial conditions and equilibrated for 5 min.
Flow Rate:0.4 mL min
Solvent A:40% water/60% acetonitrile; 10 mM ammonium formate
Solvent B:90% isopropanol/9% acetonitrile/1% water; 10 mM ammonium acetate
Chromatography Type:Reversed phase

MS:

MS ID:MS005050
Analysis ID:AN005320
Instrument Name:Agilent 6545 QTOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:For positive mode, the source gas temperature was set to 225 °C, with a drying gas flow of 11 L/minute, nebulizer pressure of 40 psig, sheath gas temp of 350 °C and sheath gas flow of 11 L/minute. VCap voltage is set at 3500 V, nozzle voltage 500V, fragmentor at 110 V, skimmer at 85 V and octopole RF peak at 750 V. For data processing, Agilent MassHunter (MH) Workstation and software packages MH Qualitiative and MH Quantitative were used. The pooled QC (n=8) and process blank (n=4) were injected throughout the sample queue to ensure the reliability of acquired lipidomics data. For lipid annotation, accurate mass and MS/MS matching was used with the Agilent Lipid Annotator library. Results from the positive and negative ionization modes from Lipid Annotator were merged based on the class of lipid identified. Data exported from MH Quantitative was evaluated using Excel where initial lipid targets are parsed based on the following criteria. Only lipids with relative standard deviations (RSD) less than 30% in QC samples are used for data analysis. Additionally, only lipids with background AUC counts in process blanks that are less than 30% of QC are used for data analysis. The parsed excel data tables are normalized based on the ratio to class-specific internal standards, then to sum prior to statistical analysis.
Ion Mode:POSITIVE
  
MS ID:MS005051
Analysis ID:AN005321
Instrument Name:Agilent 6545 QTOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:For negative mode, the source gas temperature was set to 300 °C, with a drying gas flow of 11 L/minute, a nebulizer pressure of 30 psig, sheath gas temp of 350 °C and sheath gas flow 11 L/minute. VCap voltage was set at 3500 V, nozzle voltage 75 V, fragmentor at 175 V, skimmer at 75 V and octopole RF peak at 750 V. For data processing, Agilent MassHunter (MH) Workstation and software packages MH Qualitiative and MH Quantitative were used. The pooled QC (n=8) and process blank (n=4) were injected throughout the sample queue to ensure the reliability of acquired lipidomics data. For lipid annotation, accurate mass and MS/MS matching was used with the Agilent Lipid Annotator library. Results from the positive and negative ionization modes from Lipid Annotator were merged based on the class of lipid identified. Data exported from MH Quantitative was evaluated using Excel where initial lipid targets are parsed based on the following criteria. Only lipids with relative standard deviations (RSD) less than 30% in QC samples are used for data analysis. Additionally, only lipids with background AUC counts in process blanks that are less than 30% of QC are used for data analysis. The parsed excel data tables are normalized based on the ratio to class-specific internal standards, then to sum prior to statistical analysis.
Ion Mode:NEGATIVE
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