#METABOLOMICS WORKBENCH Cloud55_20220428_003647 DATATRACK_ID:3220 STUDY_ID:ST002253 ANALYSIS_ID:AN003680 VERSION 1 CREATED_ON 03-01-2023 #PROJECT PR:PROJECT_TITLE Metabolomic profiles of T. spiralis-infected mouse serum at 0, 2, 4, 8 weeks PR:PROJECT_SUMMARY Trichinellosis is the zoonosis affected people worldwide, caused by parasitic PR:PROJECT_SUMMARY nematode in Genus Trichinella. After ingesting raw meat containing infective PR:PROJECT_SUMMARY larvae of Trichinella spp., patients may show signs of myalgia, headaches, PR:PROJECT_SUMMARY facial and periorbital edema. In severe cases, patients develop myocarditis, PR:PROJECT_SUMMARY heart failure, and possibly death. The standard method for diagnosis of PR:PROJECT_SUMMARY Trichinella infection is immunological techniques, which lack of sensitivity and PR:PROJECT_SUMMARY timeliness. Metabolomics has been extensively used to identify compounds with PR:PROJECT_SUMMARY diagnostic potential in many diseases, however, there is no study regarding PR:PROJECT_SUMMARY biomarker discovery in trichinellosis yet. Therefore, this study aims to PR:PROJECT_SUMMARY identify potential biomarkers of trichinellosis using metabolomics. Mice were PR:PROJECT_SUMMARY infected with larvae stage of T. spiralis and their serum were collected before, PR:PROJECT_SUMMARY 2 weeks, 4 weeks, and 8 weeks after infection. Metabolites in serum were PR:PROJECT_SUMMARY extracted and identified using mass spectrometer in untargeted manner. PR:PROJECT_SUMMARY Metabolomic data was annotated with XCMS online platform and analyzed with PR:PROJECT_SUMMARY Metaboanalyst version 5.0. A total of 4,688 and 5,533 metabolite features were PR:PROJECT_SUMMARY identified from positive and negative mode, respectively. The 1,139 features PR:PROJECT_SUMMARY were significantly changed metabolites and further used for pathway analysis and PR:PROJECT_SUMMARY biomarker selection. Glycerophospholipid metabolism was the major pathway PR:PROJECT_SUMMARY affected by Trichinella infection and these lipid species were the main lipid PR:PROJECT_SUMMARY class identified. The Receiver operating characteristic (ROC) revealed 247 PR:PROJECT_SUMMARY molecules with diagnostic power of trichinellosis. Phosphatidylserine was the PR:PROJECT_SUMMARY major lipid class from ROC analysis, for example, PS(12:0/15:0), PR:PROJECT_SUMMARY PS(18:0/19:0)[U]. Our study suggested glycerophospholipid and phosphatidylserine PR:PROJECT_SUMMARY species as the potential markers of trichinellosis. Findings of this study are PR:PROJECT_SUMMARY the initial step for biomarker discovery in trichinellosis, which would be a PR:PROJECT_SUMMARY benefit for improvement of disease diagnosis in the future. 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 PR:PUBLICATIONS https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0011119#pntd.0011119.ref035 PR:DOI http://dx.doi.org/10.21228/M8FX4R #STUDY ST:STUDY_TITLE Metabolomic profiles of T. spiralis-infected mouse serum at 0, 2, 4, 8 weeks ST:STUDY_SUMMARY Trichinellosis is the zoonosis affected people worldwide, caused by parasitic ST:STUDY_SUMMARY nematode in Genus Trichinella. After ingesting raw meat containing infective ST:STUDY_SUMMARY larvae of Trichinella spp., patients may show signs of myalgia, headaches, ST:STUDY_SUMMARY facial and periorbital edema. In severe cases, patients develop myocarditis, ST:STUDY_SUMMARY heart failure, and possibly death. The standard method for diagnosis of ST:STUDY_SUMMARY Trichinella infection is immunological techniques, which lack of sensitivity and ST:STUDY_SUMMARY timeliness. Metabolomics has been extensively used to identify compounds with ST:STUDY_SUMMARY diagnostic potential in many diseases, however, there is no study regarding ST:STUDY_SUMMARY biomarker discovery in trichinellosis yet. Therefore, this study aims to ST:STUDY_SUMMARY identify potential biomarkers of trichinellosis using metabolomics. Mice were ST:STUDY_SUMMARY infected with larvae stage of T. spiralis and their serum were collected before, ST:STUDY_SUMMARY 2 weeks, 4 weeks, and 8 weeks after infection. Metabolites in serum were ST:STUDY_SUMMARY extracted and identified using mass spectrometer in untargeted manner. ST:STUDY_SUMMARY Metabolomic data was annotated with XCMS online platform and analyzed with ST:STUDY_SUMMARY Metaboanalyst version 5.0. A total of 4,688 and 5,533 metabolite features were ST:STUDY_SUMMARY identified from positive and negative mode, respectively. The 1,139 features ST:STUDY_SUMMARY were significantly changed metabolites and further used for pathway analysis and ST:STUDY_SUMMARY biomarker selection. Glycerophospholipid metabolism was the major pathway ST:STUDY_SUMMARY affected by Trichinella infection and these lipid species were the main lipid ST:STUDY_SUMMARY class identified. The Receiver operating characteristic (ROC) revealed 247 ST:STUDY_SUMMARY molecules with diagnostic power of trichinellosis. Phosphatidylserine was the ST:STUDY_SUMMARY major lipid class from ROC analysis, for example, PS(12:0/15:0), ST:STUDY_SUMMARY PS(18:0/19:0)[U]. Our study suggested glycerophospholipid and phosphatidylserine ST:STUDY_SUMMARY species as the potential markers of trichinellosis. Findings of this study are ST:STUDY_SUMMARY the initial step for biomarker discovery in trichinellosis, which would be a ST:STUDY_SUMMARY benefit for improvement of disease diagnosis in the future. 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 ST:SUBMIT_DATE 2022-04-28 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 1 Experimental factor:2 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_2wk_TS2_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_2wk_TS2_1 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 2 Experimental factor:2 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_2wk_TS2_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_2wk_TS2_2 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 3 Experimental factor:2 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_2wk_TS2_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_2wk_TS2_3 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 4 Experimental factor:2 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_2wk_TS3_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_2wk_TS3_1 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 5 Experimental factor:2 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_2wk_TS3_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_2wk_TS3_2 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 6 Experimental factor:2 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_2wk_TS3_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_2wk_TS3_3 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 7 Experimental factor:2 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_2wk_TS4_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_2wk_TS4_1 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 8 Experimental factor:2 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_2wk_TS4_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_2wk_TS4_2 SUBJECT_SAMPLE_FACTORS - 2-Week post-infection 9 Experimental factor:2 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_2wk_TS4_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_2wk_TS4_3 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 1 Experimental factor:4 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_4wk_TS2_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_4wk_TS2_1 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 2 Experimental factor:4 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_4wk_TS2_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_4wk_TS2_2 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 3 Experimental factor:4 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_4wk_TS2_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_4wk_TS2_3 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 4 Experimental factor:4 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_4wk_TS3_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_4wk_TS3_1 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 5 Experimental factor:4 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_4wk_TS3_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_4wk_TS3_2 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 6 Experimental factor:4 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_4wk_TS3_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_4wk_TS3_3 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 7 Experimental factor:4 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_4wk_TS4_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_4wk_TS4_1 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 8 Experimental factor:4 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_4wk_TS4_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_4wk_TS4_2 SUBJECT_SAMPLE_FACTORS - 4-Week post-infection 9 Experimental factor:4 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_4wk_TS4_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_4wk_TS4_3 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 1 Experimental factor:8 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_8wk_TS2_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_8wk_TS2_1 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 2 Experimental factor:8 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_8wk_TS2_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_8wk_TS2_2 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 3 Experimental factor:8 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_8wk_TS2_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_8wk_TS2_3 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 4 Experimental factor:8 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_8wk_TS3_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_8wk_TS3_1 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 5 Experimental factor:8 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_8wk_TS3_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_8wk_TS3_2 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 6 Experimental factor:8 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_8wk_TS3_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_8wk_TS3_3 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 7 Experimental factor:8 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_8wk_TS4_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_8wk_TS4_1 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 8 Experimental factor:8 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_8wk_TS4_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_8wk_TS4_2 SUBJECT_SAMPLE_FACTORS - 8-Week post-infection 9 Experimental factor:8 weeks after infection RAW_FILE_NAME=20210115_Met_IDA_Pos_8wk_TS4_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_8wk_TS4_3 SUBJECT_SAMPLE_FACTORS - Control 1 Experimental factor:No infection RAW_FILE_NAME=20210115_Met_IDA_Pos_NI_TS2_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_NI_TS2_1 SUBJECT_SAMPLE_FACTORS - Control 2 Experimental factor:No infection RAW_FILE_NAME=20210115_Met_IDA_Pos_NI_TS2_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_NI_TS2_2 SUBJECT_SAMPLE_FACTORS - Control 3 Experimental factor:No infection RAW_FILE_NAME=20210115_Met_IDA_Pos_NI_TS2_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_NI_TS2_3 SUBJECT_SAMPLE_FACTORS - Control 4 Experimental factor:No infection RAW_FILE_NAME=20210115_Met_IDA_Pos_NI_TS3_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_NI_TS3_1 SUBJECT_SAMPLE_FACTORS - Control 5 Experimental factor:No infection RAW_FILE_NAME=20210115_Met_IDA_Pos_NI_TS3_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_NI_TS3_2 SUBJECT_SAMPLE_FACTORS - Control 6 Experimental factor:No infection RAW_FILE_NAME=20210115_Met_IDA_Pos_NI_TS3_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_NI_TS3_3 SUBJECT_SAMPLE_FACTORS - Control 7 Experimental factor:No infection RAW_FILE_NAME=20210115_Met_IDA_Pos_NI_TS4_1; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_NI_TS4_1 SUBJECT_SAMPLE_FACTORS - Control 8 Experimental factor:No infection RAW_FILE_NAME=20210115_Met_IDA_Pos_NI_TS4_2; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_NI_TS4_2 SUBJECT_SAMPLE_FACTORS - Control 9 Experimental factor:No infection RAW_FILE_NAME=20210115_Met_IDA_Pos_NI_TS4_3; RAW_FILE_NAME Neg=20210116_Met_IDA_Neg_NI_TS4_3 #COLLECTION CO:COLLECTION_SUMMARY 5 Mice were infected with T. spiralis 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:INSTRUMENT_NAME Agilent 1260 CH:COLUMN_NAME Waters Acquity BEH C8 (100 x 2.1mm,1.7um) CH:CHROMATOGRAPHY_TYPE Reversed phase #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 5600+ TripleTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI 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:ION_MODE POSITIVE MS:MS_RESULTS_FILE ST002253_AN003680_Results.txt UNITS:m/z Has m/z:Yes Has RT:Yes RT units:Minutes #END