#METABOLOMICS WORKBENCH Cloud55_20220331_082121 DATATRACK_ID:3150 STUDY_ID:ST002158 ANALYSIS_ID:AN003533 PROJECT_ID:PR001371 VERSION 1 CREATED_ON April 29, 2022, 1:35 am #PROJECT PR:PROJECT_TITLE Metabolomic profiles of S. mekongi-infected mouse serum at 0, 2, 4, 8 weeks PR:PROJECT_TITLE (Positive mode) PR:PROJECT_SUMMARY Serum of uninfected and S. mekongi-infected mice was collected at 2, 4, and 8 PR:PROJECT_SUMMARY weeks post-infection. Samples were extracted for metabolites and analyzed with a PR:PROJECT_SUMMARY liquid chromatography-tandem mass spectrometer. PR:INSTITUTE Princess Srisavangavadhana College of Medicine, Chulabhorn Royal Academy PR:LAST_NAME Chienwichai PR:FIRST_NAME Peerut PR:ADDRESS 906, Kamphaeng Phet 6 Rd., Lak Si, Bangkok, 10210, Thailand PR:EMAIL peerut.chi@cra.ac.th PR:PHONE +6681687460 #STUDY ST:STUDY_TITLE Untargeted serum metabolomic profiling for early detection of Schistosoma ST:STUDY_TITLE mekongi infection in mouse model ST:STUDY_SUMMARY Mekong schistosomiasis is a parasitic disease caused by blood flukes in the Lao ST:STUDY_SUMMARY People’s Democratic Republic and in Cambodia. The standard method for ST:STUDY_SUMMARY diagnosis of schistosomiasis is detection of parasite eggs from patient samples. ST:STUDY_SUMMARY However, this method is not sufficient to detect asymptomatic patients, low egg ST:STUDY_SUMMARY numbers, or early infection. Therefore, diagnostic methods with higher ST:STUDY_SUMMARY sensitivity at the early stage of the disease are needed to fill this gap. The ST:STUDY_SUMMARY aim of this study was to identify potential biomarkers of early schistosomiasis ST:STUDY_SUMMARY using an untargeted metabolomics approach. Serum of uninfected and S. ST:STUDY_SUMMARY mekongi-infected mice was collected at 2, 4, and 8 weeks post-infection. Samples ST:STUDY_SUMMARY were extracted for metabolites and analyzed with a liquid chromatography-tandem ST:STUDY_SUMMARY mass spectrometer. Metabolites were annotated with the MS-DIAL platform and ST:STUDY_SUMMARY analyzed with Metaboanalyst bioinformatic tools. Multivariate analysis ST:STUDY_SUMMARY distinguished between metabolites from the different experimental conditions. ST:STUDY_SUMMARY Biomarker screening was performed using three methods: correlation coefficient ST:STUDY_SUMMARY analysis; feature important detection with a random forest algorithm; and ST:STUDY_SUMMARY receiver operating characteristic (ROC) curve analysis. Three compounds were ST:STUDY_SUMMARY identified as potential biomarkers at the early stage of the disease: ST:STUDY_SUMMARY heptadecanoyl ethanolamide; picrotin; and theophylline. The levels of these ST:STUDY_SUMMARY three compounds changed significantly during early-stage infection, and ST:STUDY_SUMMARY therefore these molecules may be promising schistosomiasis markers. These ST:STUDY_SUMMARY findings may help to improve early diagnosis of schistosomiasis, thus reducing ST:STUDY_SUMMARY the burden on patients and limiting spread of the disease in endemic areas. ST:INSTITUTE Princess Srisavangavadhana College of Medicine, Chulabhorn Royal Academy ST:LAST_NAME Chienwichai ST:FIRST_NAME Peerut ST:ADDRESS 906, Kamphaeng Phet 6 Rd., Lak Si, Bangkok, 10210, Thailand ST:EMAIL peerut.chi@cra.ac.th ST:PHONE +6681687460 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #FACTORS #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - Control 1 Experimental factor:No infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk0_1; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk0_1 SUBJECT_SAMPLE_FACTORS - Control 2 Experimental factor:No infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk0_2; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk0_2 SUBJECT_SAMPLE_FACTORS - Control 3 Experimental factor:No infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk0_3; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk0_3 SUBJECT_SAMPLE_FACTORS - Control 4 Experimental factor:No infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk0_4; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk0_4 SUBJECT_SAMPLE_FACTORS - Control 5 Experimental factor:No infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk0_5; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk0_5 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 1 Experimental factor:2 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk14_1; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk14_1 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 2 Experimental factor:2 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk14_2; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk14_2 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 3 Experimental factor:2 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk14_3; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk14_3 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 4 Experimental factor:2 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk14_4; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk14_4 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 5 Experimental factor:2 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk14_5; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk14_5 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 1 Experimental factor:4 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk28_1; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk28_1 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 2 Experimental factor:4 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk28_2; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk28_2 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 3 Experimental factor:4 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk28_3; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk28_3 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 4 Experimental factor:4 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk28_4; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk28_4 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 5 Experimental factor:4 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk28_5; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk28_5 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 1 Experimental factor:8 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk56_1; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk56_1 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 2 Experimental factor:8 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk56_2; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk56_2 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 3 Experimental factor:8 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk56_3; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk56_3 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 4 Experimental factor:8 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk56_4; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk56_4 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 5 Experimental factor:8 weeks after infection RAW_FILE_NAME=20200831_Met_IDA_Pos_smk56_5; RAW_FILE_NAME=20200831_Met_IDA_Neg_smk56_5 #COLLECTION CO:COLLECTION_SUMMARY 5 Mice were infected with S. mekongi and serum samples were collected at 0, 2, CO:COLLECTION_SUMMARY 4, and 8 weeks after infection. Metabolite profiling was performed with mass CO:COLLECTION_SUMMARY spectrometer. CO:SAMPLE_TYPE Blood (serum) #TREATMENT TR:TREATMENT_SUMMARY Blood was collected at 0, 2,4, and 8 weeks after infection #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 20 μL serum was mixed with 80 μL cold methanol and vortexed for 1 minute. This SP:SAMPLEPREP_SUMMARY mixture was then incubated at 4°C for 20 minutes and centrifuged at 12,000 rpm SP:SAMPLEPREP_SUMMARY for 10 minutes. Next, the supernatant was collected and dried with a speed SP:SAMPLEPREP_SUMMARY vacuum (Tomy Digital Biology, Tokyo, Japan). Samples were stored at −80°C SP:SAMPLEPREP_SUMMARY until further analysis #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1260 CH:COLUMN_NAME Waters Acquity BEH C8 (100 x 2.1mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 5600+ TripleTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Information-dependent acquisition mode composed of a TOF-MS scan and 10 MS:MS_COMMENTS dependent product ion scans were used in the high sensitivity mode with dynamic MS:MS_COMMENTS background subtraction. The mass range of the TOF-MS scan was m/z 100–1,000 MS:MS_COMMENTS and the product ion scan was set to m/z 50−1,000. Equal aliquots of each MS:MS_COMMENTS metabolite sample were pooled to form the quality control (QC) samples. The QC MS:MS_COMMENTS samples were injected before, during, and after sample analysis to assess the MS:MS_COMMENTS system performance. MS:MS_RESULTS_FILE ST002158_AN003533_Results.txt UNITS:m/z Has m/z:Yes Has RT:Yes RT units:Minutes #END