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.
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.
| Study ID | ST002698 |
| Study Title | Systemic host inflammation induces stage-specific transcriptomic modification and slower maturation in malaria parasites |
| Study Type | Biomedical research |
| Study Summary | We 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 |
| 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 |
| ollie.skinner@unimelb.edu.au | |
| Phone | +61 424088268 |
| Submit Date | 2023-04-29 |
| Num Groups | 5 |
| Total Subjects | 25 |
| Num Males | NA |
| Num Females | NA |
| Study Comments | Malaria parasite cultures treatments: CpG, LPS, PbA, PIC, Saline |
| Publications | Systemic host inflammation induces stage-specific transcriptomic modification and slower maturation in malaria parasites |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-06-26 |
| Release Version | 1 |
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 |
|---|---|---|
| SA267143 | Plasma_CpG_4 | CpG |
| SA267144 | Plasma_CpG_5 | CpG |
| SA267145 | Plasma_CpG_2 | CpG |
| SA267146 | Plasma_CpG_3 | CpG |
| SA267147 | Plasma_CpG_1 | CpG |
| SA267148 | Blank_2 | Extraction blank |
| SA267149 | Blank_1 | Extraction blank |
| SA267150 | Blank_3 | Extraction blank |
| SA267151 | Plasma_LPS_5 | LPS |
| SA267152 | Plasma_LPS_4 | LPS |
| SA267153 | Plasma_LPS_2 | LPS |
| SA267154 | Plasma_LPS_3 | LPS |
| SA267155 | Plasma_LPS_1 | LPS |
| SA267161 | Plasma_PbA_3 | PbA |
| SA267162 | Plasma_PbA_5 | PbA |
| SA267163 | Plasma_PbA_2 | PbA |
| SA267164 | Plasma_PbA_1 | PbA |
| SA267165 | Plasma_PbA_4 | PbA |
| SA267156 | Plasma_PIC_1 | PIC |
| SA267157 | Plasma_PIC_3 | PIC |
| SA267158 | Plasma_PIC_4 | PIC |
| SA267159 | Plasma_PIC_2 | PIC |
| SA267160 | Plasma_PIC_5 | PIC |
| SA267166 | QC_2 | QC |
| SA267167 | QC_3 | QC |
| SA267168 | QC_1 | QC |
| SA267169 | Plasma_Sal_3 | Saline |
| SA267170 | Plasma_Sal_1 | Saline |
| SA267171 | Plasma_Sal_2 | Saline |
| SA267172 | Plasma_Sal_4 | Saline |
| SA267173 | Plasma_Sal_5 | Saline |
| 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 |
