#METABOLOMICS WORKBENCH Cloud55_20250606_201910 DATATRACK_ID:6011 STUDY_ID:ST003975 ANALYSIS_ID:AN006546 PROJECT_ID:PR002490
VERSION             	1
CREATED_ON             	June 6, 2025, 8:29 pm
#PROJECT
PR:PROJECT_TITLE                 	Metabolomic profiles of albendazole treated T. spiralis larval worms
PR:PROJECT_SUMMARY               	Trichinella spiralis is the parasitic nematodes that cause trichinellosis.
PR:PROJECT_SUMMARY               	Patients are infected with T. spiralis by consuming raw meat containing larval
PR:PROJECT_SUMMARY               	stage of the parasite. Patients may develop weakness, muscle pain, facial edema,
PR:PROJECT_SUMMARY               	or death in severe cases. Albendazole (ABZ) is the drug of choice for
PR:PROJECT_SUMMARY               	trichinellosis, however, the resistance to ABZ has been repeatedly reported.
PR:PROJECT_SUMMARY               	Hence, novel drugs against this parasite are essentially required. Developing
PR:PROJECT_SUMMARY               	new drugs is expensive and time-consuming. To overcome these limitations, the
PR:PROJECT_SUMMARY               	application of computational approaches and drug repurposing strategy would be a
PR:PROJECT_SUMMARY               	suitable solution. Here, we aimed to discover new anthelmintic drugs for T.
PR:PROJECT_SUMMARY               	spiralis using in silico-based techniques and in vitro validation. Firstly, we
PR:PROJECT_SUMMARY               	treated both adult and larval stages of T. spiralis with ABZ, then metabolome of
PR:PROJECT_SUMMARY               	the parasites were extracted. Following liquid chromatography-tandem mass
PR:PROJECT_SUMMARY               	spectrometry analysis, an approximately of 11,187-11,191 features were
PR:PROJECT_SUMMARY               	identified from XCMS online platform. From the metabolome data, there were 122
PR:PROJECT_SUMMARY               	and 133 metabolites those were significantly changed by ABZ treatment in adult
PR:PROJECT_SUMMARY               	and larval stages, respectively. The pathway analysis was performed with
PR:PROJECT_SUMMARY               	Metaboanalyst platform and fatty acid degradation pathway was highlighted in
PR:PROJECT_SUMMARY               	both stages of the parasites. This pathway is vital for lipid metabolism of the
PR:PROJECT_SUMMARY               	worms, hence, we screened for potential drug targets in this pathway. We found
PR:PROJECT_SUMMARY               	that Carnitine palmitoyl transferase (CPT) 1 and 2 are the enzymes responsible
PR:PROJECT_SUMMARY               	for transporting fatty acids into mitochondria. Inhibiting these enzymes would
PR:PROJECT_SUMMARY               	shut down the lipid metabolism machinery of the parasites. Hence, these enzymes
PR:PROJECT_SUMMARY               	were proposed as the potential drug targets for trichinellosis treatment in the
PR:PROJECT_SUMMARY               	future.
PR:INSTITUTE                     	Chulabhorn Royal Academy
PR:DEPARTMENT                    	Princess Srisavangavadhana Faculty of Medicine
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                         	+66816874608
#STUDY
ST:STUDY_TITLE                   	Metabolomic profiles of albendazole treated T. spiralis larval worms
ST:STUDY_SUMMARY                 	Trichinella spiralis is the parasitic nematodes that cause trichinellosis.
ST:STUDY_SUMMARY                 	Patients are infected with T. spiralis by consuming raw meat containing larval
ST:STUDY_SUMMARY                 	stage of the parasite. Patients may develop weakness, muscle pain, facial edema,
ST:STUDY_SUMMARY                 	or death in severe cases. Albendazole (ABZ) is the drug of choice for
ST:STUDY_SUMMARY                 	trichinellosis, however, the resistance to ABZ has been repeatedly reported.
ST:STUDY_SUMMARY                 	Hence, novel drugs against this parasite are essentially required. Developing
ST:STUDY_SUMMARY                 	new drugs is expensive and time-consuming. To overcome these limitations, the
ST:STUDY_SUMMARY                 	application of computational approaches and drug repurposing strategy would be a
ST:STUDY_SUMMARY                 	suitable solution. Here, we aimed to discover new anthelmintic drugs for T.
ST:STUDY_SUMMARY                 	spiralis using in silico-based techniques and in vitro validation. Firstly, we
ST:STUDY_SUMMARY                 	treated both adult and larval stages of T. spiralis with ABZ, then metabolome of
ST:STUDY_SUMMARY                 	the parasites were extracted. Following liquid chromatography-tandem mass
ST:STUDY_SUMMARY                 	spectrometry analysis, an approximately of 11,187-11,191 features were
ST:STUDY_SUMMARY                 	identified from XCMS online platform. From the metabolome data, there were 122
ST:STUDY_SUMMARY                 	and 133 metabolites those were significantly changed by ABZ treatment in adult
ST:STUDY_SUMMARY                 	and larval stages, respectively. The pathway analysis was performed with
ST:STUDY_SUMMARY                 	Metaboanalyst platform and fatty acid degradation pathway was highlighted in
ST:STUDY_SUMMARY                 	both stages of the parasites. This pathway is vital for lipid metabolism of the
ST:STUDY_SUMMARY                 	worms, hence, we screened for potential drug targets in this pathway. We found
ST:STUDY_SUMMARY                 	that Carnitine palmitoyl transferase (CPT) 1 and 2 are the enzymes responsible
ST:STUDY_SUMMARY                 	for transporting fatty acids into mitochondria. Inhibiting these enzymes would
ST:STUDY_SUMMARY                 	shut down the lipid metabolism machinery of the parasites. Hence, these enzymes
ST:STUDY_SUMMARY                 	were proposed as the potential drug targets for trichinellosis treatment in the
ST:STUDY_SUMMARY                 	future.
ST:INSTITUTE                     	Chulabhorn Royal Academy
ST:DEPARTMENT                    	Princess Srisavangavadhana Faculty of Medicine
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                  	Invertebrate
SU:SUBJECT_SPECIES               	Trichinella spiralis
#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           	11	20220526_Trichinella larva 0.1% DMSO_1	Sample source:T. spiralis | Stage:Larva | Treatment:Control	RAW_FILE_NAME(Raw file name)=20220526_Trichinella larva 0.1% DMSO_1
SUBJECT_SAMPLE_FACTORS           	12	20220526_Trichinella larva 0.1% DMSO_2	Sample source:T. spiralis | Stage:Larva | Treatment:Control	RAW_FILE_NAME(Raw file name)=20220526_Trichinella larva 0.1% DMSO_2
SUBJECT_SAMPLE_FACTORS           	13	20220526_Trichinella larva 0.1% DMSO_3	Sample source:T. spiralis | Stage:Larva | Treatment:Control	RAW_FILE_NAME(Raw file name)=20220526_Trichinella larva 0.1% DMSO_3
SUBJECT_SAMPLE_FACTORS           	14	20220526_Trichinella larva 0.1% DMSO_4	Sample source:T. spiralis | Stage:Larva | Treatment:Control	RAW_FILE_NAME(Raw file name)=20220526_Trichinella larva 0.1% DMSO_4
SUBJECT_SAMPLE_FACTORS           	15	20220526_Trichinella larva 0.1% DMSO_5	Sample source:T. spiralis | Stage:Larva | Treatment:Control	RAW_FILE_NAME(Raw file name)=20220526_Trichinella larva 0.1% DMSO_5
SUBJECT_SAMPLE_FACTORS           	16	20220526_Trichinella larva 100ng AB2_1	Sample source:T. spiralis | Stage:Larva | Treatment:ABZ treatment	RAW_FILE_NAME(Raw file name)=20220526_Trichinella larva 100ng AB2_1
SUBJECT_SAMPLE_FACTORS           	17	20220526_Trichinella larva 100ng AB2_2	Sample source:T. spiralis | Stage:Larva | Treatment:ABZ treatment	RAW_FILE_NAME(Raw file name)=20220526_Trichinella larva 100ng AB2_2
SUBJECT_SAMPLE_FACTORS           	18	20220526_Trichinella larva 100ng AB2_3	Sample source:T. spiralis | Stage:Larva | Treatment:ABZ treatment	RAW_FILE_NAME(Raw file name)=20220526_Trichinella larva 100ng AB2_3
SUBJECT_SAMPLE_FACTORS           	19	20220526_Trichinella larva 100ng AB2_4	Sample source:T. spiralis | Stage:Larva | Treatment:ABZ treatment	RAW_FILE_NAME(Raw file name)=20220526_Trichinella larva 100ng AB2_4
SUBJECT_SAMPLE_FACTORS           	20	20220526_Trichinella larva 100ng AB2_5	Sample source:T. spiralis | Stage:Larva | Treatment:ABZ treatment	RAW_FILE_NAME(Raw file name)=20220526_Trichinella larva 100ng AB2_5
#COLLECTION
CO:COLLECTION_SUMMARY            	Trichinella spiralis larval worms were treated with IC50 concentration of
CO:COLLECTION_SUMMARY            	albendazole, then the worms were collected for metabolite extraction
CO:SAMPLE_TYPE                   	Worms
#TREATMENT
TR:TREATMENT_SUMMARY             	Worms were treated with IC50 concentration of albendazole
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	All worms from each condition were transferred into 1.5-mL microcentrifuge tubes
SP:SAMPLEPREP_SUMMARY            	and homogenized in 500 μL methanol. At which point, the tubes were snap-frozen
SP:SAMPLEPREP_SUMMARY            	in liquid nitrogen and thawed prior to centrifuging at 800 × g for 1 min at 4°C.
SP:SAMPLEPREP_SUMMARY            	The supernatant was collected and placed in a new tube, and the pellet was
SP:SAMPLEPREP_SUMMARY            	extracted again with the same protocol. Following centrifugation, the
SP:SAMPLEPREP_SUMMARY            	supernatant from the second extraction was pooled into a tube containing the
SP:SAMPLEPREP_SUMMARY            	supernatant from the first extraction. The pellet was resuspended in 250 μL of
SP:SAMPLEPREP_SUMMARY            	deionized H2O before snap-freezing in liquid nitrogen and thawing. The
SP:SAMPLEPREP_SUMMARY            	supernatant was obtained by centrifugation at 15,000 × g for 1 min at 4°C and
SP:SAMPLEPREP_SUMMARY            	then pooled with the previous tube. The tubes containing the pooled supernatants
SP:SAMPLEPREP_SUMMARY            	were centrifuged at 15,000 × g for 1 min at 4°C to remove the remaining
SP:SAMPLEPREP_SUMMARY            	debris. The clear supernatant was transferred to a new tube and later dried in a
SP:SAMPLEPREP_SUMMARY            	speed vacuum.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 1260
CH:COLUMN_NAME                   	Waters Acquity BEH C8 (100 x 2.1 mm, 1.7 μm)
CH:SOLVENT_A                     	100% Water; 0.1% Formic acid
CH:SOLVENT_B                     	100% Acetonitrile; 0.1% Formic acid
CH:FLOW_GRADIENT                 	5% mobile phase B for 2.0 min (0.0–2.0 min), 5%–60% B for 0.5 min (2.0–2.5
CH:FLOW_GRADIENT                 	min), 60%–80% B for 1.5 min (2.5–4.0 min), 80%–100% B for 8.0 min
CH:FLOW_GRADIENT                 	(4.0–12.0 min), 100% B for 5 min (12.0–17.0 min), 100%–5% B for 0.1 min
CH:FLOW_GRADIENT                 	(17.0–17.1 min), and 5% B for 2.9 min (17.1–20.0 min).
CH:FLOW_RATE                     	0.3 mL/min
CH:COLUMN_TEMPERATURE            	40°C
#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               	ST003975_AN006546_Results.txt	UNITS:m/z	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END