Summary of Study ST002549

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.

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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 IDST002549
Study TitleLipidomic analysis of serum from mice with Toxoplasma gondii infection
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 Date2023-04-07
Raw Data AvailableYes
Raw Data File Type(s)raw(Waters)
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

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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:SU002649
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 Factor
SA256069Toxoplasma infected-5Infected
SA256070Toxoplasma infected-4Infected
SA256071Toxoplasma infected-3Infected
SA256072Toxoplasma infected-6Infected
SA256073Toxoplasma infected-7Infected
SA256074Toxoplasma infected-9Infected
SA256075Toxoplasma infected-8Infected
SA256076Toxoplasma infected-2Infected
SA256077Toxoplasma infected-1Infected
SA256078Uninfected-4Uninfected
SA256079Uninfected-3Uninfected
SA256080Uninfected-2Uninfected
SA256081Uninfected-5Uninfected
SA256082Uninfected-6Uninfected
SA256083Uninfected-9Uninfected
SA256084Uninfected-8Uninfected
SA256085Uninfected-7Uninfected
SA256086Uninfected-1Uninfected
Showing results 1 to 18 of 18

Collection:

Collection ID:CO002642
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 (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.
Sample Type:Blood (serum)

Treatment:

Treatment ID:TR002661
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:SP002655
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 AN004197
Analysis type MS
Chromatography type Reversed phase
Chromatography system Waters Acquity
Column Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name Waters Xevo TQ-S
Ion Mode UNSPECIFIED
Units pmol/mL of plasma

Chromatography:

Chromatography ID:CH003110
Chromatography Summary:Low pH polar
Instrument Name:Waters Acquity
Column Name:Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
Column Temperature:65
Flow Gradient:30% B for 0.5min, a linear gradient to 40% B for 3min, 60% B over 3min, then 85% B for 4.5min
Flow Rate:0.4mL/min
Solvent A:60% water/40% acetonitrile
Solvent B:90% isopropanol/10% methanol
Chromatography Type:Reversed phase

MS:

MS ID:MS003944
Analysis ID:AN004197
Instrument Name:Waters Xevo TQ-S
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Lipid extraction and analysis for targeted sphingolipid analysis was done using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS)69. Lipids were extracted from sera using an azeotrophic mix of isopropanol:water:ethyl acetate (3:1:6; v:v:v). Internal standards (10 pmol of d17 long-chain bases and C12 acylated sphingolipids) were added to samples at the onset of the extraction procedure. Extracts were separated on a Waters Iclass Acquity UPLC chromatography system. Mobile phases were (A) 60:40 water:acetonitrile and (B) 90:10 isopropanol:methanol with both mobile phases containing 5 mM ammonium formate and 0.1% formic acid. A Waters C18 CSH 2.1 mm ID × 10 cm column maintained at 65°C was used for the separation of the sphingoid bases, 1-phosphates, and acylated sphingolipids. The eluate was analyzed with an inline Waters TQ-S mass spectrometer using multiple reaction monitoring.
Ion Mode:UNSPECIFIED
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