Summary of Study ST002939

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 PR001828. The data can be accessed directly via it's Project DOI: 10.21228/M8F14S 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 IDST002939
Study TitleRole of PI3K in Atrial Myopathy: Insights from Transgenic Mouse Models and Identification of a Dysregulated PI3K Lipid Profile in Individuals with Atrial Fibrillation - Part 2 of 2, Mus musculus
Study SummaryLipidomic profiling was conducted on atrias and ventricles from 20 week old, male and female caPI3K and dnPI3K heterozygous and homozygous transgenic mice. These transgenic mice have cardiac specific overexpression of either a constitutively active PI3K transgene (to have increased PI3K activity in the heart, mimicking an athlete's heart) or dominate negative PI3K transgene (to have decreased PI3K activity in the heart, resulting in smaller hearts with increase susceptibility to a variety of cardiac insults).
Institute
Baker Heart and Diabetes Institute
LaboratoryMetabolomics
Last NameTham
First NameYow
Address75 Commercial Rd
Emailyowkeat.tham@baker.edu.au
Phone0430502623
Submit Date2023-10-17
Num Groups6
Total Subjects105
Num Males46
Num Females59
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2025-01-12
Release Version1
Yow Tham Yow Tham
https://dx.doi.org/10.21228/M8F14S
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:


Project:

Project ID:PR001828
Project DOI:doi: 10.21228/M8F14S
Project Title:Role of PI3K in Atrial Myopathy: Insights from Transgenic Mouse Models and Identification of a Dysregulated PI3K Lipid Profile in Individuals with Atrial Fibrillation
Project Summary:In a serendipitous discovery, atrial enlargement, fibrosis and thrombi was identified in a subset of transgenic mice with reduced phosphoinositide 3-kinase (PI3K, class IA) in cardiac myocytes. Understanding mechanisms underlying atrial myopathy has important implications for understanding and preventing atrial fibrillation (AF). Prior work had shown that PI3K is an essential regulator of exercise-induced ventricular enlargement and protection, but the role in the atria was unknown. Further, while targeting IGF1-PI3K-Akt signaling has been considered a potential therapeutic strategy for the failing heart, growing evidence suggests fine-tuning IGF1-PI3K signaling would be necessary. Here, we undertook comprehensive physiological and molecular analyses in cardiac-specific transgenic mice with increased or decreased PI3K to assess the dose response impact of directly regulating PI3K. Elevated PI3K was associated with a dose-dependent increase in heart size, and preserved/enhanced function. In contrast, reduced PI3K led to cardiac dysfunction, fibrosis, arrhythmia, and increased susceptibility to atrial enlargement and thrombi. This phenotype was associated with dysregulation of a lipid species (GM3) that regulates the IGF1-PI3K pathway, cardiac stress and contractility genes. Proteomic profiling identified distinct signatures across atria with varying degrees of atrial dysfunction, enlargement, and presence of atrial thrombi. To assess the potential relevance in humans we assessed circulating PI3K-related lipids in plasma from athletes with/without AF. Dysregulation of GM3 and PI3K-related lipids were identified in athletes with AF. Collectively, this work advances our understanding of mechanisms underpinning atrial pathophysiology, offers new insights for therapeutic approaches targeting atrial myopathy and AF, and has identified potential new lipid markers for identifying individuals at risk of AF.
Institute:Baker Heart and Diabetes Institute
Laboratory:Metabolomics
Last Name:Tham
First Name:Yow Keat
Address:75 Commercial Rd, Melbourne, Victoria, 3004, Australia
Email:yowkeat.tham@baker.edu.au
Phone:0430502623

Subject:

Subject ID:SU003052
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:FVBN
Age Or Age Range:approx 20 weeks old
Gender:Male and female
Animal Feed:Specialty Feeds Irradiated Rat and Mouse Standard Chow Diet
Species Group:Mammals

Factors:

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

mb_sample_id local_sample_id Tissue Sex Genotype
SA31949039027_AAtria Female caPI3K Tg (+/+)
SA31949138965_AAtria Female caPI3K Tg (+/+)
SA31949238845_AAtria Female caPI3K Tg (+/+)
SA31949338852_AAtria Female caPI3K Tg (+/+)
SA31949438853_AAtria Female caPI3K Tg (+/+)
SA31949538843_AAtria Female caPI3K Tg (+/+)
SA31949638917_AAtria Female caPI3K Tg (+/+)
SA31949739002_AAtria Female caPI3K Tg (+/+)
SA31949838967_AAtria Female caPI3K Tg (+/+)
SA31949938966_AAtria Female caPI3K Tg (+/+)
SA31950039003_AAtria Female caPI3K Tg (+/+)
SA31950138968_AAtria Female caPI3K Tg (+/+)
SA31950238860_AAtria Female caPI3K Tg (+/-)
SA31950338854_AAtria Female caPI3K Tg (+/-)
SA31950439005_AAtria Female caPI3K Tg (+/-)
SA31950538914_AAtria Female caPI3K Tg (+/-)
SA31950638915_AAtria Female caPI3K Tg (+/-)
SA31950739004_AAtria Female caPI3K Tg (+/-)
SA31950838969_AAtria Female caPI3K Tg (+/-)
SA31950939026_AAtria Female caPI3K Tg (+/-)
SA31951038859_AAtria Female caPI3K Tg (+/-)
SA31951138881_AAtria Female dnPI3K Tg (+/+)
SA31951238998_AAtria Female dnPI3K Tg (+/+)
SA31951338999_AAtria Female dnPI3K Tg (+/+)
SA31951439028_AAtria Female dnPI3K Tg (+/+)
SA31951539035_AAtria Female dnPI3K Tg (+/+)
SA31951639030_AAtria Female dnPI3K Tg (+/+)
SA31951739036_AAtria Female dnPI3K Tg (+/+)
SA31951838879_AAtria Female dnPI3K Tg (+/+)
SA31951938877_AAtria Female dnPI3K Tg (+/+)
SA31952038880_AAtria Female dnPI3K Tg (+/+)
SA31952139032_AAtria Female dnPI3K Tg (+/-)
SA31952238871_AAtria Female dnPI3K Tg (+/-)
SA31952338878_AAtria Female dnPI3K Tg (+/-)
SA31952438872_AAtria Female dnPI3K Tg (+/-)
SA31952538992_AAtria Female dnPI3K Tg (+/-)
SA31952638997_AAtria Female dnPI3K Tg (+/-)
SA31952738990_AAtria Female dnPI3K Tg (+/-)
SA31952838991_AAtria Female dnPI3K Tg (+/-)
SA31952938867_AAtria Female dnPI3K Tg (+/-)
SA31953039033_AAtria Female dnPI3K Tg (+/-)
SA31947339029_AAtria Female Non-transgenic
SA31947439000_AAtria Female Non-transgenic
SA31947538996_AAtria Female Non-transgenic
SA31947638993_AAtria Female Non-transgenic
SA31947739034_AAtria Female Non-transgenic
SA31947838869_AAtria Female Non-transgenic
SA31947938870_AAtria Female Non-transgenic
SA31948038995_AAtria Female Non-transgenic
SA31948139031_AAtria Female Non-transgenic
SA31948238994_AAtria Female Non-transgenic
SA31948338912_AAtria Female Non-transgenic
SA31948438842_AAtria Female Non-transgenic
SA31948538851_AAtria Female Non-transgenic
SA31948638913_AAtria Female Non-transgenic
SA31948739001_AAtria Female Non-transgenic
SA31948838916_AAtria Female Non-transgenic
SA31948938964_AAtria Female Non-transgenic
SA31954138858_AAtria Male caPI3K Tg (+/+)
SA31954238970_AAtria Male caPI3K Tg (+/+)
SA31954338855_AAtria Male caPI3K Tg (+/+)
SA31954438856_AAtria Male caPI3K Tg (+/+)
SA31954539009_AAtria Male caPI3K Tg (+/+)
SA31954638848_AAtria Male caPI3K Tg (+/+)
SA31954738849_AAtria Male caPI3K Tg (+/+)
SA31954839008_AAtria Male caPI3K Tg (+/+)
SA31954938863_AAtria Male caPI3K Tg (+/-)
SA31955038864_AAtria Male caPI3K Tg (+/-)
SA31955139006_AAtria Male caPI3K Tg (+/-)
SA31955239007_AAtria Male caPI3K Tg (+/-)
SA31955338857_AAtria Male caPI3K Tg (+/-)
SA31955438973_AAtria Male caPI3K Tg (+/-)
SA31955538971_AAtria Male caPI3K Tg (+/-)
SA31955638847_AAtria Male caPI3K Tg (+/-)
SA31955738846_AAtria Male caPI3K Tg (+/-)
SA31955839077_AAtria Male dnPI3K Tg (+/+)
SA31955939076_AAtria Male dnPI3K Tg (+/+)
SA31956039085_AAtria Male dnPI3K Tg (+/+)
SA31956139139_AAtria Male dnPI3K Tg (+/+)
SA31956239071_AAtria Male dnPI3K Tg (+/+)
SA31956339142_AAtria Male dnPI3K Tg (+/+)
SA31956439087_AAtria Male dnPI3K Tg (+/+)
SA31956539072_AAtria Male dnPI3K Tg (+/-)
SA31956639074_AAtria Male dnPI3K Tg (+/-)
SA31956739073_AAtria Male dnPI3K Tg (+/-)
SA31956838884_AAtria Male dnPI3K Tg (+/-)
SA31956938882_AAtria Male dnPI3K Tg (+/-)
SA31957039078_AAtria Male dnPI3K Tg (+/-)
SA31957139084_AAtria Male dnPI3K Tg (+/-)
SA31957239140_AAtria Male dnPI3K Tg (+/-)
SA31957339088_AAtria Male dnPI3K Tg (+/-)
SA31957439086_AAtria Male dnPI3K Tg (+/-)
SA31957539141_AAtria Male dnPI3K Tg (+/-)
SA31957639070_AAtria Male dnPI3K Tg (+/-)
SA31953139075_AAtria Male Non-transgenic
SA31953238886_AAtria Male Non-transgenic
SA31953338883_AAtria Male Non-transgenic
SA31953439079_AAtria Male Non-transgenic
SA31953538885_AAtria Male Non-transgenic
SA31953638866_AAtria Male Non-transgenic
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Collection:

Collection ID:CO003045
Collection Summary:Ventricles were dissected from mice such that each sample will include left and right ventricular tissue. Ventricles and atrias were snap frozen in liquid nitrogen and stored in -80 freezer until tissues were processed for lipid extractions
Sample Type:Ventricles and Atria
Storage Conditions:-80℃

Treatment:

Treatment ID:TR003061
Treatment Summary:Mice did not undergo specific treatment, as this was a basal phenotyping study. Mice were fasted for 6 hours before dissections, and a lethal dose of anesthesia was delivered via intraperitoneal injection before tissue collection.
Animal Anesthesia:Pentobarbitone
Animal Fasting:6 hours

Sample Preparation:

Sampleprep ID:SP003058
Sampleprep Summary:Tissues were homogenised in 1xPBS and then sonicated with a probe-sonicator for 15 seconds, 23 amplitude. BCA assays were then conducted to determine protein concentrations of these homogenates. Lipid extraction was conducted using 10ul of sample (ventricle homogenate at 5mg/ml, atria homogenate at 2.5mg/ml) using the single phase chloroform methanol method. 10ul of internal standards and 200ul of chloroform:methanol (1:2) were added to samples before the mixture was vortexed. Samples were then placed on a rotary shaker for 10 mins at a speed of 90 before being transferred to a bath sonicator. Samples were then sonicated for 30 mins at water temperature below 28 degrees. Samples were then removed and rested at room temperature for 20 mins. Samples were then centrifuged at 13000rpm for 10 minutes. 200ul of the supernatant was then transferred to 0.5ml polypropylene 96 well plates, and spun dried using a speedvac vacuum concentrator. Lipids were reconstituted in 50ul water saturated butanol + 50ul of Ammonium Formate.
Processing Storage Conditions:On ice
Extract Storage:-80℃

Combined analysis:

Analysis ID AN004824
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/µmol of Total Phosphatidylcholine

Chromatography:

Chromatography ID:CH003644
Chromatography Summary:The running solvent consisted of solvent A: 50% H2O / 30% acetonitrile / 20% isopropanol (v/v/v) containing 10mM ammonium formate and 5uM medronic acid, and solvent B: 1% H2O / 9% acetonitrile / 90% isopropanol (v/v/v) containing 10mM ammonium formate. We utilized a stepped linear gradient with a 16-minute cycle time per sample and a 1µL sample injection. To increase throughput, we used a dual column set up to equilibrate the second column while the first is running a sample. The sample analytical gradient was as follows: starting with a flow rate of 0.4mL/minute at 15% B and increasing to 50% B over 2.5 minutes, then to 57% over 0.1 minutes, to 70% over 6.4 minutes, to 93% over 0.1 minute, to 96% over 1.9 minutes and finally to 100% over 0.1 minute. The solvent was then held at 100% B for 0.9 minutes (total 12.0 minutes). Equilibration was started as follows: solvent was decreased from 100% B to 15% B over 0.2 minutes and held until a total of 16 minutes. The next sample is injected and the columns are switched.
Instrument Name:Agilent 1290 Infinity II
Column Name:Agilent ZORBAX Eclipse Plus C18 (100 x 2.1mm,1.8um)
Column Temperature:45
Flow Gradient:Starting with a flow rate of 0.4mL/minute at 15% B and increasing to 50% B over 2.5 minutes, then to 57% over 0.1 minutes, to 70% over 6.4 minutes, to 93% over 0.1 minute, to 96% over 1.9 minutes and finally to 100% over 0.1 minute. The solvent was then held at 100% B for 0.9 minutes (total 12.0 minutes). Equilibration was started as follows: solvent was decreased from 100% B to 15% B over 0.2 minutes and held until a total of 16 minutes.
Flow Rate:0.4mL/min
Solvent A:50% water/30% acetonitrile/20% isopropanol; 10mM ammonium formate; 5uM medronic acid
Solvent B:1% water/9% acetonitrile/90% isopropanol; 10mM ammonium formate
Chromatography Type:Reversed phase

MS:

MS ID:MS004570
Analysis ID:AN004824
Instrument Name:Agilent 6490 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Details previously published in https://doi.org/10.1016/j.chembiol.2018.10.008 Analysis of plasma extracts was performed on an Agilent 6490 QQQ mass spectrometer with an Agilent 1290 series HPLC system and a ZORBAX eclipse plus C18 column (2.1x100mm 1.8μm, Agilent) with the thermostat set at 60°C. Mass spectrometry analysis was performed in positive ion mode with dynamic scheduled multiple reaction monitoring (MRM). Mass spectrometry settings and MRM transitions for each lipid class, subclass and individual species are shown in Tables 1 and S1. The solvent system consisted of solvent A) 50% H2O / 30% acetonitrile / 20% isopropanol (v/v/v) containing 10mM ammonium formate and solvent B) 1% H2O / 9% acetonitrile / 90% isopropanol (v/v/v) containing 10mM ammonium formate. We utilized a stepped linear gradient with a 15-minute cycle time per sample and a 1μL sample injection. The gradient was as follows; starting with a flow rate of 0.4ml/minute at 10% B and increasing to 45% B over 2.7 minutes, then to 53% over 0.1 minutes, to 65% over 6.2 minutes, to 89% over 0.1 minute, to 92% over 1.9 minutes and finally to 100% over 0.1 minute. The solvent was then held at 100% B for 0.8 minutes (total 11.9 minutes). Equilibration was as follows, solvent was decreased from 100% B to 10% B over 0.1 minute and held for an additional 0.9 minutes. Flow rate was then switched to 0.6 ml/minute for 1 minute before returning to 0.4 ml/minute over 0.1 minutes. Solvent B was held at 10% B for a further 0.9 minutes at 0.4ml/minutes for a total cycle time of 15 minutes. The following mass spectrometer conditions were used; gas temperature, 150°C, gas flow rate 17L/min, nebulizer 20psi, Sheath gas temperature 200°C, capillary voltage 3500V and sheath gas flow 10L/min. Isolation widths for Q1 and Q3 were set to “unit” resolution (0.7 amu). PQC samples consisting of a pooled set of 6 healthy individuals were incorporated into the analysis at 1 PQC per 18 plasma samples. TQC consisted of PQC extracts which were pooled and split into individual vials to provide a measure of technical variation from the mass spectrometer only. These were included at a ratio of 1 TQC per 18 plasma samples. TQCs were monitored for changes in peak area, width and retention time to determine the performance of the LC-MS/MS analysis and were subsequently used to align for differential responses across the analytical batches.
Ion Mode:POSITIVE
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