Summary of Study ST003168

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 PR001927. The data can be accessed directly via it's Project DOI: 10.21228/M8N42X 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 IDST003168
Study TitleLipidomic analysis of cryopreserved human cardiac tissue from young and ageing adults
Study SummaryUntargeted lipidomic analysis was performed to measure lipids in left ventricular heart tissue from pre-mortem healthy donor hearts, as classified by formal pathological examination. Hearts were stored at the Sydney Heart Bank. Samples were divided into young (age ≤ 25 years) and old (age ≥ 50 years) cohorts. Lipidomic analysis used liquid chromatography-tandem mass spectrometry (LC-MS/MS) on a high resolution Q-Exactive HF-X Quadrupole-Orbitrap mass spectrometer, operated in both positive and negative ionisation mode.
Institute
University of Sydney
DepartmentMedicine and Health
LaboratoryLipid Metabolism and Neurochemistry
Last NameDon
First NameAnthony
AddressThe Hub, Charles Perkins Centre, D17, The University of Sydney, NSW, 2006
Emailanthony.don@sydney.edu.au
Phone+61 2 8627 5578
Submit Date2024-03-20
Num Groups2
Total Subjects23
Num Males15
Num Females8
Raw Data AvailableYes
Raw Data File Type(s)mzML
Analysis Type DetailLC-MS
Release Date2024-05-03
Release Version1
Anthony Don Anthony Don
https://dx.doi.org/10.21228/M8N42X
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:


Project:

Project ID:PR001927
Project DOI:doi: 10.21228/M8N42X
Project Title:The Human Cardiac “Age-OME”: Age-specific changes in myocardial molecular expression
Project Type:MS analysis
Project Summary:A substantial proportion of the World’s population is ageing. One of the most significant risk factors for heart disease is ageing. However, we do not understand how the human heart changes with age. Taking advantage of a unique set of pre-mortem, cryopreserved, non-diseased human hearts, we performed multi-omic analyses (transcriptomics, proteomics, metabolomics and lipidomics), coupled with biological computational modelling in younger (<25 years old) and older hearts (>50years old) to describe the molecular landscape of human cardiac ageing. In older hearts, we observed a downregulation of proteins involved in calcium signalling and of the contractile apparatus itself. In addition, we found a potential counteractive upregulation of central carbon generation of fuel, upregulation of glycolysis and increases in long-chain fatty acids. This is the first molecular data set of normal human cardiac ageing, which may have important implications for the development of age-related heart disease.
Institute:University of Sydney
Department:School of Medical Sciences
Laboratory:Cardiometabolic Medicine
Last Name:Koay
First Name:Yen Chin
Address:The Hub, Charles Perkins Centre, D17, The University of Sydney, NSW, 2006
Email:yen.koay@sydney.edu.au
Phone:+61486275851

Subject:

Subject ID:SU003287
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Age Or Age Range:Young donors (aged 25 years or younger) and older donors (aged 50 years or older).
Gender:Male and female

Factors:

Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Sample source Sex Age group
SA343041old_5Heart F old
SA343042old_1Heart F old
SA343043old_7Heart F old
SA343044old_11Heart F old
SA343045old_13Heart F old
SA343046young_10Heart F young
SA343047young_12Heart F young
SA343048young_7Heart F young
SA343049old_10Heart M old
SA343050old_8Heart M old
SA343051old_4Heart M old
SA343052old_2Heart M old
SA343053old_12Heart M old
SA343054old_9Heart M old
SA343055young_3Heart M young
SA343056young_4Heart M young
SA343057young_2Heart M young
SA343058young_6Heart M young
SA343059young_8Heart M young
SA343060young_11Heart M young
SA343061young_1Heart M young
SA343062young_9Heart M young
SA343063young_5Heart M young
Showing results 1 to 23 of 23

Collection:

Collection ID:CO003280
Collection Summary:Control donor hearts that were considered suitable for transplantation but ultimately not used due to factors such as transport issues, immune mismatches, and size discrepancies between donor and recipient, were collected in this study. These hearts came from individuals who died of non-cardiac reasons and had no risk factors for heart disease, including having a BMI under 30. Following a thorough pathological review, these hearts were confirmed to be structurally normal through histological evaluation. Left ventricular (LV) samples were taken directly in the operating room, immediately snap-frozen in liquid nitrogen at -196°C, and then stored at the Sydney Heart Bank at the University of Sydney, at temperatures ranging from -170 to -180°C. This collection process received ethical approval from the University of Sydney's Ethics Committee (USYD # 2021/122).
Collection Protocol ID:CO003212
Sample Type:Heart
Storage Conditions:Described in summary

Treatment:

Treatment ID:TR003296
Treatment Summary:The samples detailed in this study did not undergo any treatment.
Treatment Protocol ID:TR003228

Sample Preparation:

Sampleprep ID:SP003294
Sampleprep Summary:Lipids were extracted from ~20 mg heart tissue using a two-phase method with methyl-tert-butyl ether (MTBE) and water. Tissue was homogenised with steel beads in 250 µL methanol containing 0.01% (w/v) butylhydroxytoluene (BHT) and mass spectrometry internal standards: 2 nmoles PC(19:0/19:0); 1 nmole each of SM(d18:1/12:0), GluCer(d18:1/12:0), Cer(d18:1/17:0), PS(17:0/17:0), PE(17:0/17:0), PA(17:0/17:0), PI(d7-18:1/15:0), PG(17:0/17:0), CL(14:0/14:0/14:0/14:0), and TG(17:0/17:0/17:0); 0.5 nmoles each of DG(d7-18:1/15:0), CholE(17:0), LPC (17:0), LPE(17:1), and AcCa(d3-16:0); and 0.2 nmole each of Sph(d17:1), S1P(d17:1), LacCer(d18:1/12:0), and MG(d7-18:1). MTBE (850 µL) was added, and samples were sonicated in a 4°C water bath for 30 min. Mass spectrometry grade water (212 µL) was added to induce phase separation after vortexing and centrifugation at 2000g for 5 min. The upper organic phase was collected in 5 mL glass tubes and the aqueous phase was extracted twice more with 500 µL MTBE and 150 µL methanol followed by sonication for 15 min and phase separation with 125 µL water. Organic phases from the three extractions were combined and dried under vacuum in a Savant SC210 SpeedVac (ThermoFisher Scientific). Lipids were reconstituted in 400 µL 80% methanol/20% water/0.1% formic acid containing 0.01% (w/v) BHT and stored at -80 °C.
Sampleprep Protocol ID:SP003225

Combined analysis:

Analysis ID AN005198 AN005199
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Thermo Vanquish Thermo Vanquish
Column Waters HPLC C18 (100 x 2.1mm, 1.7um) Waters HPLC C18 (100 x 2.1mm, 1.7um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive HF-X Orbitrap Thermo Q Exactive HF-X Orbitrap
Ion Mode POSITIVE NEGATIVE
Units pmoles/mg tissue nanomoles/mg of tissue

Chromatography:

Chromatography ID:CH003933
Chromatography Summary:A ThermoFisher Q-Exactive HF-X mass spectrometer coupled to a Vanquish HPLC with a Waters Acquity UPLC BEH C18 (100 x 2.1 mm, 1.7 um) column was used for LC-MS/MS, with minor modifications to our previously-reported method. HPLC solvent A was 10 mM ammonium formate, 0.1% formic acid in acetonitrile:water (60:40), and solvent B was 10 mM ammonium formate, 0.1% formic acid in isopropanol:acetonitrile (90:10). A 27 min binary gradient at 0.28 mL/min was used: 0 min, 80:20 A/B; 3 min, 80:20 A/B; 5.5 min, 55:45 A/B; 8 min, 35:65 A/B; 13 min, 15:85 A/B; 14 min, 0:100 A/B; 20 min, 0:100 A/B; 20.2 min, 70:30 A/B; 27 min, 70:30 A/B.
Instrument Name:Thermo Vanquish
Column Name:Waters HPLC C18 (100 x 2.1mm, 1.7um)
Column Temperature:32
Flow Gradient:0 min, 80:20 A/B; 3 min, 80:20 A/B; 5.5 min, 55:45 A/B; 8 min, 35:65 A/B; 13 min, 15:85 A/B; 14 min, 0:100 A/B; 20 min, 0:100 A/B; 20.2 min, 70:30 A/B; 27 min, 70:30 A/B
Flow Rate:0.28 mL/min
Solvent A:60% acetonitrile/40% water; 10 mM ammonium formate; 0.1% formic acid
Solvent B:90% isopropanol/10% acetonitrile; 10 mM ammonium formate; 0.1% formic acid
Chromatography Type:Reversed phase

MS:

MS ID:MS004931
Analysis ID:AN005198
Instrument Name:Thermo Q Exactive HF-X Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Data was acquired in full scan/data-dependent MS2 mode (full scan resolution 60,000 FWHM, scan range 220–1600 m/z). Sample order was randomised, and data was collected in both positive and negative mode for each sample. The ten most abundant ions in each cycle were subjected to MS2, with an isolation window of 1.4 m/z, collision energy 30 eV, resolution 17,500 FWHM, maximum integration time 110 ms and dynamic exclusion window 10 s. An exclusion list of background ions was used based on a solvent blank. An inclusion list was used for all internal standards. LipidSearch software (version 4.2, Thermo Fisher) was used for lipid annotation, chromatogram alignment, and peak integration. Lipid annotation required both accurate precursor ion mass (tolerance 5 ppm) and diagnostic product ions (tolerance 8 ppm). Individual lipids were expressed as ratios to the class-specific internal standard, then multiplied by the amount of internal standard to calculate molar amounts for each lipid. Lipid levels were expressed as nmoles/mg tissue.
Ion Mode:POSITIVE
  
MS ID:MS004932
Analysis ID:AN005199
Instrument Name:Thermo Q Exactive HF-X Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Data was acquired in full scan/data-dependent MS2 mode (full scan resolution 60,000 FWHM, scan range 220–1600 m/z). Sample order was randomised, and data was collected in both positive and negative mode for each sample. The ten most abundant ions in each cycle were subjected to MS2, with an isolation window of 1.4 m/z, collision energy 30 eV, resolution 17,500 FWHM, maximum integration time 110 ms and dynamic exclusion window 10 s. An exclusion list of background ions was used based on a solvent blank. An inclusion list was used for all internal standards. LipidSearch software (version 4.2, Thermo Fisher) was used for lipid annotation, chromatogram alignment, and peak integration. Lipid annotation required both accurate precursor ion mass (tolerance 5 ppm) and diagnostic product ions (tolerance 8 ppm). Individual lipids were expressed as ratios to the class-specific internal standard, then multiplied by the amount of internal standard to calculate molar amounts for each lipid. Lipid levels were expressed as nmoles/mg tissue.
Ion Mode:NEGATIVE
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