Summary of Study ST002698

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 PR001670. The data can be accessed directly via it's Project DOI: 10.21228/M8V14H 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 IDST002698
Study TitleSystemic host inflammation induces stage-specific transcriptomic modification and slower maturation in malaria parasites
Study TypeBiomedical research
Study SummaryWe found that host inflammation altered the plasma environment surrounding Plasmodium falciparum parasites in vivo, and that this altered plasma environment contained inhibitory factors that directly impaired maturation of early trophozoite stages. We demonstrated with LPS-conditioning that systemic host inflammation alone, in the absence of confounding factors such as ongoing infection, slowed the rate at which parasites transited from one generation of RBC to the next. While this is consistent with the idea that host inflammatory responses can impair parasite maturation, other TLR agonists, CpG and Poly I:C, did not elicit such a response. Metabolomics also identified 1-methylhypoxanthine as elevated in both LPS conditioned and acutely-infected plasma. Plasmodium survival depends on host hypoxanthine, inosine and xanthine for purine synthesis. 1-Methylhypoxanthine can bind effectively to and possibly limit the action of hypoxanthine-guanine phosphoribosyl transferase (HGPRTase)25, an enzyme critical for purine synthesis. Interestingly, hypoxanthine, inosine and xanthine were also all reduced in the plasma of LPS-conditioned and acutely infected mice supporting the possibility that inhibition of purine synthesis by 1-methylhypoxanthine might have been partly aided by the lack of substrates for this pathway.
Institute
Peter Doherty Institute for Infection and Immunity
DepartmentDepartment of Microbiology and Immunology
LaboratoryAshraful Haque lab
Last NameSkinner
First NameOliver
Address792 Elizabeth Street, The University of Melbourne, Victoria 3000 Australia
Emailollie.skinner@unimelb.edu.au
Phone+61 424088268
Submit Date2023-04-29
Num Groups5
Total Subjects25
Num MalesNA
Num FemalesNA
Study CommentsMalaria parasite cultures treatments: CpG, LPS, PbA, PIC, Saline
PublicationsSystemic host inflammation induces stage-specific transcriptomic modification and slower maturation in malaria parasites
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2023-06-26
Release Version1
Oliver Skinner Oliver Skinner
https://dx.doi.org/10.21228/M8V14H
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR001670
Project DOI:doi: 10.21228/M8V14H
Project Title:Systemic host inflammation induces stage-specific transcriptomic modification and slower maturation in malaria parasites (University of Melbourne)
Project Type:Untargeted LCMS metabolomics
Project Summary:Maturation rates of malaria parasites within red blood cells (RBC) can be influenced by host nutrient status and circadian rhythm; whether host inflammatory responses can also influence maturation remains less clear. Here, we observed that systemic host inflammation induced in mice by an innate immune stimulus, lipopolysaccharide (LPS), or by ongoing acute Plasmodium infection, slowed the progression of a single cohort of parasites from one generation of RBC to the next. Importantly, plasma from LPS-conditioned or acutely-infected mice directly inhibited parasite maturation during in vitro culture, which was not rescued by supplementation, suggesting the emergence of inhibitory factors in plasma. Metabolomic assessments confirmed substantial alterations to the plasma of LPS-conditioned and acutely-infected mice, and identified a small number of candidate inhibitory metabolites, some of which could interfere with Plasmodium purine synthesis. Finally, we confirmed rapid parasite responses to systemic host inflammation in vivo using parasite scRNA-seq, noting broad impairment in transcriptional activity and translational capacity specifically in trophozoites, but not rings or schizonts. Thus, we provide evidence that systemic host inflammation rapidly triggered transcriptional alterations in circulating blood-stage Plasmodium trophozoites, and predict candidate inhibitory metabolites in the plasma that may impair parasite maturation in vivo.
Institute:Peter Doherty Institute for Infection and Immunity
Department:Department of Microbiology and Immunology
Laboratory:Ashraful Haque lab
Last Name:Skinner
First Name:Oliver
Address:792 Elizabeth Street, The University of Melbourne, Victoria 3000 Australia
Email:ollie.skinner@unimelb.edu.au
Phone:+61 424088268
Publications:Systemic host inflammation induces stage-specific transcriptomic modification and slower maturation in malaria parasites
Contributors:Lianne I.M. Lansink, Oliver P. Skinner, Jessica A. Engel, Hyun Jae Lee, Megan S.F. Soon, Cameron G. Williams, Arya SheelaNair, Clara P.S. Pernold, Pawat Laohamonthonkul, Jasmin Akter, Thomas Stoll, Michelle Hill, Arthur M. Talman, Andrew Russell, Mara Lawniczak, Xiaoxiao Jia, Brendon Chua, Dovile Anderson, Darren J. Creek, Miles P. Davenport, David S. Khoury, Ashraful Haque

Subject:

Subject ID:SU002801
Subject Type:Other organism
Subject Species:Plasmodium berghei
Taxonomy ID:5823
Genotype Strain:ANKA
Age Or Age Range:NA
Weight Or Weight Range:NA
Height Or Height Range:NA
Gender:Not applicable

Factors:

Subject type: Other organism; Subject species: Plasmodium berghei (Factor headings shown in green)

mb_sample_id local_sample_id Extraction blank
SA267143Plasma_CpG_4CpG
SA267144Plasma_CpG_5CpG
SA267145Plasma_CpG_2CpG
SA267146Plasma_CpG_3CpG
SA267147Plasma_CpG_1CpG
SA267148Blank_2Extraction blank
SA267149Blank_1Extraction blank
SA267150Blank_3Extraction blank
SA267151Plasma_LPS_5LPS
SA267152Plasma_LPS_4LPS
SA267153Plasma_LPS_2LPS
SA267154Plasma_LPS_3LPS
SA267155Plasma_LPS_1LPS
SA267161Plasma_PbA_3PbA
SA267162Plasma_PbA_5PbA
SA267163Plasma_PbA_2PbA
SA267164Plasma_PbA_1PbA
SA267165Plasma_PbA_4PbA
SA267156Plasma_PIC_1PIC
SA267157Plasma_PIC_3PIC
SA267158Plasma_PIC_4PIC
SA267159Plasma_PIC_2PIC
SA267160Plasma_PIC_5PIC
SA267166QC_2QC
SA267167QC_3QC
SA267168QC_1QC
SA267169Plasma_Sal_3Saline
SA267170Plasma_Sal_1Saline
SA267171Plasma_Sal_2Saline
SA267172Plasma_Sal_4Saline
SA267173Plasma_Sal_5Saline
Showing results 1 to 31 of 31

Collection:

Collection ID:CO002794
Collection Summary:C57BL/6J mice were purchased from the Animal Resource Centre (Perth, Australia). C57BL/6J.rag1−/− mice were bred at QIMR Berghofer Medical Research Institute. All mice were female between 6-12 weeks of age and were maintained under conventional conditions. Plasmodium berghei ANKA parasites constitutively expressing high-levels of eGFP (for RBC adoptive transfer), or luciferase (for establishing acute infection, although bioluminescence was not utilised), were sourced and used as previously reported12,16,17. PbA parasites were used after defrosting cryopreserved infected blood and a single in vivo passage in C57BL/6J mice. RBCs were collected from passage mice by cardiac puncture and used to infect with 105 pRBCs via lateral tail vein injection.
Sample Type:infected Red Blood Cells
Storage Conditions:-80℃

Treatment:

Treatment ID:TR002810
Treatment Summary:Mice were treated with Saline (0.9%; Baxter), Lipopolysaccharide (LPS; 0.75 mg/mL) (Sigma-Aldrich), CpG 1826 (0.25 mg/mL) (Sigma-Aldrich) or Polyriboinosinic:polyribocytidylic acid (Poly I:C; 2 mg/mL) (InvivoGen) via intraperitoneal injection (200 μL per mouse) using 26G needles, two hours prior to adoptive transfer of CFTR-labelled RBCs.

Sample Preparation:

Sampleprep ID:SP002807
Sampleprep Summary:Two independent experiments were conducted, each with 6 mice per treatment group. Ice-cold butanol/methanol (1:1) containing 50 µg/mL antioxidant 2,6-di-tert-butyl-4-methylphenol (BHT) was added to each plasma sample, as well as a pooled quality control (QC) sample at 10x volume. Samples were vortexed for 10 seconds then snap frozen on dry ice. Thawed samples were sonicated for 15 minutes on ice, stored for 2 hours at -30°C, and then centrifuged for 15 minutes at 16,000×g (4°C). Supernatant was collected, aliquoted, dried down using a vacuum concentrator and stored at -80°C until LC/MS analysis.
Processing Storage Conditions:4℃
Extraction Method:MeOH/BuOH (1:1)
Extract Storage:-80℃
Sample Resuspension:NA
Sample Derivatization:NA

Combined analysis:

Analysis ID AN004372 AN004373
Analysis type MS MS
Chromatography type HILIC HILIC
Chromatography system Thermo Dionex Ultimate 3000 RS Thermo Dionex Ultimate 3000 RS
Column SeQuant ZIC-HILIC (150 x 4.6mm,3.5um) SeQuant ZIC-HILIC (150 x 4.6mm,3.5um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive Orbitrap Thermo Q Exactive Orbitrap
Ion Mode POSITIVE NEGATIVE
Units peak height peak height

Chromatography:

Chromatography ID:CH003278
Chromatography Summary:pHILIC chromatography pH9, 32 min run with posneg switching
Methods Filename:Metabolomics_pHILIC_Parkville_v1.pdf
Instrument Name:Thermo Dionex Ultimate 3000 RS
Column Name:SeQuant ZIC-HILIC (150 x 4.6mm,3.5um)
Column Pressure:60 bar at starting conditions. 180 bar at %A
Column Temperature:25 C
Flow Gradient:0 min - 80%B, 15 min - 50%B, 18 min - 5%B, 21 min - 5%B, 24 min - 80%B, 32 min - 80%B
Flow Rate:0.3 ml/min
Injection Temperature:4 C
Sample Injection:10 uL
Solvent A:20 mM ammonium carbonate
Solvent B:acetonitrile
Analytical Time:32 min
Capillary Voltage:4 kV
Oven Temperature:25 C
Washing Buffer:syringe wash 50% IPA
Weak Wash Solvent Name:50% IPA
Strong Wash Volume:50% IPA
Sample Loop Size:25 uL
Sample Syringe Size:25 uL
Randomization Order:yes
Chromatography Type:HILIC

MS:

MS ID:MS004120
Analysis ID:AN004372
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Acquired using XCalibur software (Thermo Scientific)
Ion Mode:POSITIVE
Capillary Temperature:300 C
Capillary Voltage:4 kV
Collision Energy:NA
Collision Gas:NA
Dry Gas Flow:50
Dry Gas Temp:120
Ion Source Temperature:120 C
Ionization:ESI
Mass Accuracy:3 ppm
Precursor Type:[M+H]+
Acquisition Parameters File:Metabolomics_pHILIC_Parkville_v1.pdf
Analysis Protocol File:PQMS3-MPMF-WIN-0501_LCMS_data_acquisition_for_untargeted_metabolomics_analysis.pdf
  
MS ID:MS004121
Analysis ID:AN004373
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Acquired using XCalibur software (Thermo Scientific)
Ion Mode:NEGATIVE
Capillary Temperature:300 C
Capillary Voltage:3.5 kV
Collision Energy:NA
Collision Gas:NA
Dry Gas Flow:50
Dry Gas Temp:120
Ion Source Temperature:120 C
Ionization:ESI
Mass Accuracy:3 ppm
Precursor Type:[M-H]-
Acquisition Parameters File:Metabolomics_pHILIC_Parkville_v1.pdf
Analysis Protocol File:PQMS3-MPMF-WIN-0501_LCMS_data_acquisition_for_untargeted_metabolomics_analysis.pdf
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