Untargeted serum metabolomics analysis of Trichinella spiralis-infected mouse

Abstract

Background Trichinellosis, caused by a parasitic nematode of the genus Trichinella, is a zoonosis that affects people worldwide. After ingesting raw meat containing Trichinella spp. larvae, patients show signs of myalgia, headaches, and facial and periorbital edema, and severe cases may die from myocarditis and heart failure. The molecular mechanisms of trichinellosis are unclear, and the sensitivity of the diagnostic methods used for this disease are unsatisfactory. Metabolomics is an excellent tool for studying disease progression and biomarkers; however, it has never been applied to trichinellosis. We aimed to elucidate the impacts of Trichinella infection on the host body and identify potential biomarkers using metabolomics. Methodology/Principal findings Mice were infected with T. spiralis larvae, and sera were collected before and 2, 4, and 8 weeks after infection. Metabolites in the sera were extracted and identified using untargeted mass spectrometry. Metabolomic data were annotated via the XCMS online platform and analyzed with Metaboanalyst version 5.0. A total of 10,221 metabolomic features were identified, and the levels of 566, 330, and 418 features were significantly changed at 2-, 4-, and 8-weeks post-infection, respectively. The altered metabolites were used for further pathway analysis and biomarker selection. A major pathway affected by Trichinella infection was glycerophospholipid metabolism, and glycerophospholipids comprised the main metabolite class identified. Receiver operating characteristic revealed 244 molecules with diagnostic power for trichinellosis, with phosphatidylserines (PS) being the primary lipid class. Some lipid molecules, e.g., PS (18:0/19:0)[U] and PA (O-16:0/21:0), were not present in metabolome databases of humans and mice, thus they may have been secreted by the parasites. Conclusions/Significance Our study highlighted glycerophospholipid metabolism as the major pathway affected by trichinellosis, hence glycerophospholipid species are potential markers of trichinellosis. The findings of this study represent the initial steps in biomarker discovery that may benefit future trichinellosis diagnosis.

Metabolite profiling of Trichinella spiralis adult worms and muscle larvae identifies their excretory and secretory products

“…According to our previous study, ( Chienwichai et al., 2023 ) the metabolomics analysis of T. spiralis -infected mouse sera has been reported. We found that D-glucose 6-phosphat found in adult worms and larvae was also detected in the T. spiralis -infected mouse sera at 4 weeks post-infection.…”

“…The metabolome extraction was performed, according to the previous study ( Chienwichai et al., 2023 ). For ES product, 20 µL of solution was mixed with 80 µL of cold methanol using vortex for 1 min.…”

“…The metabolome was identified using mass spectrometry, according to the previous study ( Chienwichai et al., 2023 ). Briefly, metabolome were identified using ultra high-performance liquid chromatography (UHPLC; Agilent 1260 Quaternary Pump, Agilent 1260 High Performance Autosampler and Agilent 1290 Thermostatted Column Compartment SL, Agilent Technologies, CA, USA) combined with a quadrupole time-of-flight mass spectrometer (Q-TOF MS; TripleTOF 5600+, SCIEX, US).…”

“…Although some previous research have been conducted to identify biomarkers specific to T. spiralis , most of them were focused on the protein-based biomarkers ( Wang et al., 2013 ; Wang et al., 2014 ; Thawornkuno et al., 2022 ). On the other hand, only one study worked on the identification of metabolite-based biomarker from infected mice sera has been reported ( Chienwichai et al., 2023 ). During the infection, parasitic nematodes release many excretory-secretory (ES) products with the immunomodulatory activity in order to escape host immune system.…”

“…It is applicable in many research areas, including biomarker discovery and drug development ( Wishart, 2016 ; Alseekh et al., 2021 ). To date, few metabolite profiles of T. spiralis have become available ( Chienwichai et al., 2023 ). Consequently, the stage-specific metabolomes of T. spiralis adult worms and muscle larvae were characterized in this study using mass spectrometry-based metabolomics and bioinformatics, which also identified metabolite-based biomarkers from their excretory–secretory (ES) products.…”