{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002698","ANALYSIS_ID":"AN004372","VERSION":"1","CREATED_ON":"12-21-2023"},

"PROJECT":{"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","DOI":"http://dx.doi.org/10.21228/M8V14H","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"},

"STUDY":{"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","EMAIL":"ollie.skinner@unimelb.edu.au","PHONE":"+61 424088268","SUBMIT_DATE":"2023-04-29"},

"SUBJECT":{"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"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"Plasma_CpG_1",
"Factors":{"Extraction blank":"CpG"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_CpG_1"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_CpG_2",
"Factors":{"Extraction blank":"CpG"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_CpG_2"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_CpG_3",
"Factors":{"Extraction blank":"CpG"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_CpG_3"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_CpG_4",
"Factors":{"Extraction blank":"CpG"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_CpG_4"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_CpG_5",
"Factors":{"Extraction blank":"CpG"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_CpG_5"}
},
{
"Subject ID":"-",
"Sample ID":"Blank_1",
"Factors":{"Extraction blank":"Extraction blank"},
"Additional sample data":{"RAW_FILE_NAME":"Blank_1"}
},
{
"Subject ID":"-",
"Sample ID":"Blank_2",
"Factors":{"Extraction blank":"Extraction blank"},
"Additional sample data":{"RAW_FILE_NAME":"Blank_2"}
},
{
"Subject ID":"-",
"Sample ID":"Blank_3",
"Factors":{"Extraction blank":"Extraction blank"},
"Additional sample data":{"RAW_FILE_NAME":"Blank_3"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_LPS_1",
"Factors":{"Extraction blank":"LPS"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_LPS_1"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_LPS_2",
"Factors":{"Extraction blank":"LPS"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_LPS_2"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_LPS_3",
"Factors":{"Extraction blank":"LPS"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_LPS_3"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_LPS_4",
"Factors":{"Extraction blank":"LPS"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_LPS_4"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_LPS_5",
"Factors":{"Extraction blank":"LPS"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_LPS_5"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_PbA_1",
"Factors":{"Extraction blank":"PbA"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_PbA_1"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_PbA_2",
"Factors":{"Extraction blank":"PbA"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_PbA_2"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_PbA_3",
"Factors":{"Extraction blank":"PbA"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_PbA_3"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_PbA_4",
"Factors":{"Extraction blank":"PbA"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_PbA_4"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_PbA_5",
"Factors":{"Extraction blank":"PbA"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_PbA_5"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_PIC_1",
"Factors":{"Extraction blank":"PIC"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_PIC_1"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_PIC_2",
"Factors":{"Extraction blank":"PIC"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_PIC_2"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_PIC_3",
"Factors":{"Extraction blank":"PIC"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_PIC_3"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_PIC_4",
"Factors":{"Extraction blank":"PIC"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_PIC_4"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_PIC_5",
"Factors":{"Extraction blank":"PIC"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_PIC_5"}
},
{
"Subject ID":"-",
"Sample ID":"QC_1",
"Factors":{"Extraction blank":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC_1"}
},
{
"Subject ID":"-",
"Sample ID":"QC_2",
"Factors":{"Extraction blank":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC_2"}
},
{
"Subject ID":"-",
"Sample ID":"QC_3",
"Factors":{"Extraction blank":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC_3"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_Sal_1",
"Factors":{"Extraction blank":"Saline"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_Sal_1"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_Sal_2",
"Factors":{"Extraction blank":"Saline"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_Sal_2"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_Sal_3",
"Factors":{"Extraction blank":"Saline"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_Sal_3"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_Sal_4",
"Factors":{"Extraction blank":"Saline"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_Sal_4"}
},
{
"Subject ID":"-",
"Sample ID":"Plasma_Sal_5",
"Factors":{"Extraction blank":"Saline"},
"Additional sample data":{"RAW_FILE_NAME":"Plasma_Sal_5"}
}
],
"COLLECTION":{"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_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."},

"SAMPLEPREP":{"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"},

"CHROMATOGRAPHY":{"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"},

"ANALYSIS":{"LABORATORY_NAME":"Monash Proteomics and Metabolomics Platform","ANALYSIS_TYPE":"MS","ANALYSIS_PROTOCOL_FILE":"PQMS3-MPMF-WIN-0501_LCMS_data_acquisition_for_untargeted_metabolomics_analysis.pdf","ACQUISITION_DATE":"10/11/2022","ACQUISITION_PARAMETERS_FILE":"Metabolomics_pHILIC_Parkville_v1.pdf","SOFTWARE_VERSION":"XCalibur 4.6","OPERATOR_NAME":"Dovile Anderson","DETECTOR_TYPE":"Full scan","DATA_FORMAT":".RAW"},

"MS":{"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]+"},

"MS_METABOLITE_DATA":{
"Units":"peak height",

"Data":[{"Metabolite":"11-amino-undecanoic acid","Plasma_CpG_1":"844706304.0000","Plasma_CpG_2":"1948374784.0000","Plasma_CpG_3":"1166285696.0000","Plasma_CpG_4":"937341504.0000","Plasma_CpG_5":"920687232.0000","Blank_1":"825930.0000","Blank_2":"811318.0000","Blank_3":"770625.0000","Plasma_LPS_1":"614660672.0000","Plasma_LPS_2":"1168460416.0000","Plasma_LPS_3":"869150272.0000","Plasma_LPS_4":"801944512.0000","Plasma_LPS_5":"756473792.0000","Plasma_PbA_1":"2546736640.0000","Plasma_PbA_2":"1268040064.0000","Plasma_PbA_3":"1558644352.0000","Plasma_PbA_4":"1932263424.0000","Plasma_PbA_5":"1310396672.0000","Plasma_PIC_1":"926067712.0000","Plasma_PIC_2":"1046043584.0000","Plasma_PIC_3":"1162321408.0000","Plasma_PIC_4":"1034487296.0000","Plasma_PIC_5":"852074432.0000","QC_1":"1215249152.0000","QC_2":"1155080448.0000","QC_3":"1123049344.0000","Plasma_Sal_1":"1037037504.0000","Plasma_Sal_2":"916866496.0000","Plasma_Sal_3":"882346624.0000","Plasma_Sal_4":"1430008192.0000","Plasma_Sal_5":"831293056.0000"},{"Metabolite":"1_6_8-trimethylallantoate","Plasma_CpG_1":"971289.0000","Plasma_CpG_2":"967554.0000","Plasma_CpG_3":"1354826.0000","Plasma_CpG_4":"1318628.0000","Plasma_CpG_5":"1258573.0000","Blank_1":"","Blank_2":"","Blank_3":"","Plasma_LPS_1":"680129.0000","Plasma_LPS_2":"654027.0000","Plasma_LPS_3":"541614.0000","Plasma_LPS_4":"493495.0000","Plasma_LPS_5":"645549.0000","Plasma_PbA_1":"502987.0000","Plasma_PbA_2":"621112.0000","Plasma_PbA_3":"755580.0000","Plasma_PbA_4":"679655.0000","Plasma_PbA_5":"748900.0000","Plasma_PIC_1":"1076323.0000","Plasma_PIC_2":"821643.0000","Plasma_PIC_3":"719059.0000","Plasma_PIC_4":"917313.0000","Plasma_PIC_5":"954310.0000","QC_1":"951059.0000","QC_2":"927769.0000","QC_3":"856111.0000","Plasma_Sal_1":"1370478.0000","Plasma_Sal_2":"1455119.0000","Plasma_Sal_3":"1427286.0000","Plasma_Sal_4":"1104550.0000","Plasma_Sal_5":"1236876.0000"},{"Metabolite":"1-Methylhistidine","Plasma_CpG_1":"18314952.0000","Plasma_CpG_2":"21158194.0000","Plasma_CpG_3":"19892316.0000","Plasma_CpG_4":"19600950.0000","Plasma_CpG_5":"17859132.0000","Blank_1":"","Blank_2":"","Blank_3":"","Plasma_LPS_1":"29429098.0000","Plasma_LPS_2":"30139314.0000","Plasma_LPS_3":"48895556.0000","Plasma_LPS_4":"26523378.0000","Plasma_LPS_5":"32269530.0000","Plasma_PbA_1":"28344616.0000","Plasma_PbA_2":"36723960.0000","Plasma_PbA_3":"37741204.0000","Plasma_PbA_4":"31470626.0000","Plasma_PbA_5":"37451472.0000","Plasma_PIC_1":"36963592.0000","Plasma_PIC_2":"26419532.0000","Plasma_PIC_3":"27840620.0000","Plasma_PIC_4":"30896132.0000","Plasma_PIC_5":"25997192.0000","QC_1":"57919824.0000","QC_2":"53646508.0000","QC_3":"55003904.0000","Plasma_Sal_1":"27718144.0000","Plasma_Sal_2":"17567616.0000","Plasma_Sal_3":"17731634.0000","Plasma_Sal_4":"16947416.0000","Plasma_Sal_5":"15153468.0000"},{"Metabolite":"1-Methylhypoxanthine","Plasma_CpG_1":"235359.0000","Plasma_CpG_2":"309615.0000","Plasma_CpG_3":"462142.0000","Plasma_CpG_4":"396895.0000","Plasma_CpG_5":"315613.0000","Blank_1":"24533.0000","Blank_2":"44598.0000","Blank_3":"74129.0000","Plasma_LPS_1":"2167706.0000","Plasma_LPS_2":"1894581.0000","Plasma_LPS_3":"1623351.0000","Plasma_LPS_4":"755397.0000","Plasma_LPS_5":"689809.0000","Plasma_PbA_1":"1909127.0000","Plasma_PbA_2":"1694488.0000","Plasma_PbA_3":"1984566.0000","Plasma_PbA_4":"2323452.0000","Plasma_PbA_5":"1691930.0000","Plasma_PIC_1":"471651.0000","Plasma_PIC_2":"933921.0000","Plasma_PIC_3":"738160.0000","Plasma_PIC_4":"961154.0000","Plasma_PIC_5":"369264.0000","QC_1":"1283708.0000","QC_2":"1581831.0000","QC_3":"1055225.0000","Plasma_Sal_1":"918359.0000","Plasma_Sal_2":"721800.0000","Plasma_Sal_3":"473266.0000","Plasma_Sal_4":"561897.0000","Plasma_Sal_5":"1111245.0000"},{"Metabolite":"(1R_2R)-2-amino-1-(4-aminophenyl)propane-1_3-diol","Plasma_CpG_1":"767476.0000","Plasma_CpG_2":"433473.0000","Plasma_CpG_3":"928602.0000","Plasma_CpG_4":"477898.0000","Plasma_CpG_5":"932416.0000","Blank_1":"125341.0000","Blank_2":"161041.0000","Blank_3":"152740.0000","Plasma_LPS_1":"3420647.0000","Plasma_LPS_2":"4948794.0000","Plasma_LPS_3":"5183142.0000","Plasma_LPS_4":"4241025.0000","Plasma_LPS_5":"3898429.0000","Plasma_PbA_1":"622694.0000","Plasma_PbA_2":"667768.0000","Plasma_PbA_3":"1115391.0000","Plasma_PbA_4":"1052822.0000","Plasma_PbA_5":"806058.0000","Plasma_PIC_1":"2189106.0000","Plasma_PIC_2":"2051477.0000","Plasma_PIC_3":"2098585.0000","Plasma_PIC_4":"1764820.0000","Plasma_PIC_5":"1562343.0000","QC_1":"2196630.0000","QC_2":"2141997.0000","QC_3":"1707423.0000","Plasma_Sal_1":"813287.0000","Plasma_Sal_2":"462439.0000","Plasma_Sal_3":"1189949.0000","Plasma_Sal_4":"705992.0000","Plasma_Sal_5":"870260.0000"},{"Metabolite":"1-(sn-Glycero-3-phospho)-myo-inositol","Plasma_CpG_1":"56172.0000","Plasma_CpG_2":"62892.0000","Plasma_CpG_3":"64353.0000","Plasma_CpG_4":"59655.0000","Plasma_CpG_5":"86200.0000","Blank_1":"","Blank_2":"","Blank_3":"","Plasma_LPS_1":"104935.0000","Plasma_LPS_2":"166517.0000","Plasma_LPS_3":"133028.0000","Plasma_LPS_4":"121445.0000","Plasma_LPS_5":"107145.0000","Plasma_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