Summary of Study ST002545

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 PR001639. The data can be accessed directly via it's Project DOI: 10.21228/M8V41D 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.

Perform statistical analysis  |  Show all samples  |  Show named metabolites  |  Download named metabolite data  |  Perform analysis on untargeted data  
Download mwTab file (text)   |  Download mwTab file(JSON)   |  Download data files (Contains raw data)
Study IDST002545
Study TitleLipidomic profile of Toxoplasma gondii-infected mice (Positive mode MS)
Study SummaryCachexia is a life-threatening disease characterized by chronic, inflammatory muscle wasting and systemic metabolic impairment. Despite its high prevalence, there are no efficacious therapies for cachexia. Mice chronically infected with the protozoan parasite Toxoplasma gondii represent a novel animal model recapitulating the chronic kinetics of cachexia. To understand how perturbations to metabolic tissue homeostasis influence circulating metabolite availability we used mass spectrometry analysis. Despite the significant reduction in circulating triacylglycerides, nonesterified fatty acids, and glycerol, sphingolipid long-chain bases and a subset of phosphatidylcholines (PCs) were significantly increased in the sera of mice with T. gondii infection-induced cachexia. In addition, the TCA cycle intermediates -ketoglutarate, 2- hydroxyglutarate, succinate, fumarate, and malate were highly depleted in cachectic mouse sera. Sphingolipids and their de novo synthesis precursors PCs are the major components of the mitochondrial membrane and regulate mitochondrial function consistent with a causal relationship in the energy imbalance driving T. gondii-induced chronic cachexia.
Institute
University of Virginia
Last NameFeng
First NameTzu-Yu
Address345 Crispell Dr.
Emailttf4ye@virginia.edu
Phone702-217-4454
Submit Date2023-04-06
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2023-04-20
Release Version1
Tzu-Yu Feng Tzu-Yu Feng
https://dx.doi.org/10.21228/M8V41D
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:


Project:

Project ID:PR001639
Project DOI:doi: 10.21228/M8V41D
Project Title:Metabolomic study on the chronic Toxoplasma gondii infection in mice.
Project Type:Untargeted MS
Project Summary:Cachexia is a life-threatening disease characterized by chronic, inflammatory muscle wasting and systemic metabolic impairment. Despite its high prevalence, there are no efficacious therapies for cachexia. Mice chronically infected with the protozoan parasite Toxoplasma gondii represent a novel animal model recapitulating the chronic kinetics of cachexia. To understand how perturbations to metabolic tissue homeostasis influence circulating metabolite availability we used mass spectrometry analysis. Despite the significant reduction in circulating triacylglycerides, nonesterified fatty acids, and glycerol, sphingolipid long-chain bases and a subset of phosphatidylcholines (PCs) were significantly increased in the sera of mice with T. gondii infection-induced cachexia. In addition, the TCA cycle intermediates alpha-ketoglutarate, 2- hydroxyglutarate, succinate, fumarate, and malate were highly depleted in cachectic mouse sera. Sphingolipids and their de novo synthesis precursors PCs are the major components of the mitochondrial membrane and regulate mitochondrial function consistent with a causal relationship in the energy imbalance driving T. gondii-induced chronic cachexia.
Institute:University of Virginia
Last Name:Feng
First Name:Tzu-Yu
Address:345 Crispell Dr.
Email:ttf4ye@virginia.edu
Phone:702-217-4454

Subject:

Subject ID:SU002645
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

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

mb_sample_id local_sample_id Factor
SA255642Toxoplasma infected-4Infected
SA255643Toxoplasma infected-7Infected
SA255644Toxoplasma infected-3Infected
SA255645Toxoplasma infected-2Infected
SA255646Toxoplasma infected-6Infected
SA255647Toxoplasma infected-5Infected
SA255648Toxoplasma infected-1Infected
SA255649Toxoplasma infected-8Infected
SA255650Uninfected-6Uninfected
SA255651Uninfected-4Uninfected
SA255652Uninfected-8Uninfected
SA255653Uninfected-3Uninfected
SA255654Uninfected-2Uninfected
SA255655Uninfected-5Uninfected
SA255656Uninfected-7Uninfected
SA255657Uninfected-1Uninfected
Showing results 1 to 16 of 16

Collection:

Collection ID:CO002638
Collection Summary:At 7 weeks post-infection, isoflurane-anesthetized mice were retro-orbitally bled and sera was flash frozen and sent to the National Institute of Health (NIH) West Coast Metabolomics Center (UC Davis) for untargeted mass spectrometry analysisusing the primary metabolism assay (ALEXCIS GCTOF-MS) or the complex lipids (CSH-QTOF MS) assay. Detected meatbolites were identified based on retention time and mass spectra from MassBank of North America, curated by the NIH West Coast Metabolomics Center , and reported as raw peak heights. The raw peak heights from each analytical platform were normalized to the average peak heights of the identified metabolites in uninfected group. The resulting data were analyzed for fold-change and multiple unpaired t-test and visualized using volcano plots to identify the differential expression of metabolites in response to T. gondii-induced cachexia
Sample Type:Blood (serum)

Treatment:

Treatment ID:TR002657
Treatment Summary:To generate cysts for infection, 8-10 week female CBA/J mice were infected with 10 Me49 bradyzoite cysts by intraperitoneal injection. 4–8 weeks following infection, mice were euthanized by CO2 inhalation and cysts were harvest from brains homogenate passed through a 70 μm filter. Homegenate was washed 3 times in PBS, stained with dolichos biflorus agglutinin conjugated to FITC (Vector labs) at a 1:500 dilution. The number of cysts were determined by counting FITCpositive cysts at 20x magnification using an EVOS FL imaging system (Thermo Fisher). For experimental infections 10–14-week-old male C57BL/6 mice were infected with 10 Me49 bradyzoite cysts by intraperitoneal infection resuspended in 200 Μl PBS per mouse using a 5G 5/8” tuberculin syringe. Prior to infection, mice were cross-housed on dirty, wood chip bedding for two weeks to normalize commensal microbiota and limit the effect of eating corn husk bedding on dietary metabolites. At experimental endpoints, mice were fasted for 4 hours and isoflurane anaesthetized to isolate sera via retro-orbital bleed and/or euthanized by CO2 asphyxiation to harvest tissues for weighing and histological analysis.

Sample Preparation:

Sampleprep ID:SP002651
Sampleprep Summary:At 7 weeks post-infection, isoflurane-anesthetized mice were retro-orbitally bled and sera was flash frozen and sent to the National Institute of Health (NIH) West Coast Metabolomics Center (UC Davis) for untargeted mass spectrometry analysisusing the primary metabolism assay (ALEXCIS GCTOF-MS) or the complex lipids (CSH-QTOF MS) assay. Detected meatbolites were identified based on retention time and mass spectra from MassBank of North America, curated by the NIH West Coast Metabolomics Center , and reported as raw peak heights (Table S1 and S3). The raw peak heights from each analytical platform were normalized to the average peak heights of the identified metabolites in uninfected group. The resulting data were analyzed for fold-change and multiple unpaired t-test and visualized using volcano plots to identify the differential expression of metabolites in response to T. gondii-induced cachexia

Combined analysis:

Analysis ID AN004192
Analysis type MS
Chromatography type Reversed phase
Chromatography system Agilent 1290 Infinity
Column Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
MS Type ESI
MS instrument type QTOF
MS instrument name Agilent 6530 QTOF
Ion Mode POSITIVE
Units reads

Chromatography:

Chromatography ID:CH003106
Instrument Name:Agilent 1290 Infinity
Column Name:Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
Column Temperature:65
Flow Gradient:15%-99%
Flow Rate:0.6 ml/min
Solvent A:60% water/40% acetonitrile; 5 mM ammonium formate; 0.1% formic acid
Solvent B:90% isopropanol/10% methanol; 5 mM ammonium formate; 0.1% formic acid
Chromatography Type:Reversed phase

MS:

MS ID:MS003939
Analysis ID:AN004192
Instrument Name:Agilent 6530 QTOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:Samples extracted using Matyash extraction procedure which includes MTBE, MeOH, and H2O. The organic (upper) phase was dried down and submitted for resuspension and injection onto the LC while the aqueous (bottom) phase was dried down and submitted to derivatization for GC. They are resuspended with 110 uL of a solution of 9:1 methanol: toluene and 50 ng/mL CUDA. This is then shaken for 20 seconds, sonicated for 5 minutes at room temperature, and then centrifuged for 2 minutes at 16100 rcf. The samples are then aliquoted into three parts. 33 uL are aliquoted into a vial with a 50 uL glass insert for positive and negative mode lipidomics. The last part is aliquoted into an eppendorf tube to be used as a pool. The samples are then loaded up on an Agilent 1290 Infinity LC stack. The positive and negative mode were run on an Agilent 6530 with a scan range of m/z 120-1200 Da with an acquisition speed of 2 spectra/s. Between 0.5 and 2 uL were injected onto a Waters ACQUITY UPLC CSH C18 1.7um 2.1x100mm Column with VanGuard PreColumn 2.1x5 mm . The gradient used is 0 min 15% (B), 0–2 min 30% (B), 2–2.5 min 48% (B), 2.5–11 min 82% (B), 11–11.5 min 99% (B), 11.5–12 min 99% (B), 12–12.1 min 15% (B), 12.1–15 min 15% (B) with a flow rate of 0.6 mL/min. The mass resolution for the Agilent 6530 is 10,000 for ESI (+).
Ion Mode:POSITIVE
  logo