Comparative Metabonomic Investigations of Schistosoma japonicum From SCID Mice and BALB/c Mice: Clues to Developmental Abnormality of Schistosome in the Immunodeficient Host - PubMed (original) (raw)
Comparative Metabonomic Investigations of Schistosoma japonicum From SCID Mice and BALB/c Mice: Clues to Developmental Abnormality of Schistosome in the Immunodeficient Host
Rong Liu et al. Front Microbiol. 2019.
Abstract
The growth and development of schistosome has been affected in the immunodeficient hosts. But it remains unresolved about the molecular mechanisms involved in the development and reproduction regulation of schistosomes. This study tested and compared the metabolic profiles of the male and female Schistosoma japonicum worms collected from SCID mice and BALB/c mice at 5 weeks post infection using liquid chromatography tandem mass spectrometry (LC-MS/MS) platform, in which the worms from SCID mice were the investigated organisms and the worms from BALB/c mice were used as the controls. There were 1015 ion features in ESI+ mode and 342 ion features in ESI- mode were identified after filtration by false discovery rate. Distinct metabolic profiles were found to clearly differentiate both male and female worms in SCID mice from those in BALB/c mice using multivariate modeling methods including the Principal Component Analysis (PCA), Partial Least Squares Discriminant Analysis (PLS-DA), and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA). There were more differential metabolites in female worms than in male worms between SCID mice and BALB/c mice. And common and uniquely perturbed metabolites and pathways were identified among male and female worms from SCID mice when compared with BALB/c mice. The enriched metabolite sets of the differential metabolites in male worms between SCID mice and BALB/c mice included bile acid biosynthesis, taurine and hypotaurine metabolism, sphingolipid metabolism, retinol metabolism, purine metabolism, etc. And the enriched metabolite sets of differential metabolites in female worms included retinol metabolism, alpha linolenic acid and linoleic acid metabolism, purine metabolism, sphingolipid metabolism, glutamate metabolism, etc. Further detection and comparison in transcript abundance of genes of the perturbed retinol metabolism and its associated meiosis process in worms identified clues suggesting accumulated retinyl ester and perturbed meiotic process. These findings suggested an association between the schistosome with retarded growth and development in SCID mice and their perturbed metabolites and metabolic pathways, and provided a new insight into the growth and development regulation of S. japonicum worms from the metabolic level, which indicated great clues for discovery of drugs or vaccines against the parasites and disease with more researches.
Keywords: BALB/c mouse; LC-MS/MS; SCID mouse; Schistosoma japonicum; growth and development; metabolomics.
Figures
FIGURE 1
Differential metabolic profiles of male and female S. japonicum worms between SCID mice and BALB/c mice at 35 days post infection. Principal Component Analysis (PCA) score scatter plots of metabolites obtained from LC-MS/MS fingerprints in ESI+ (A) and ESI- mode (B). Partial least-squares discriminant analysis (PLS-DA) separating metabolites of the four groups of worms and QC sample in ESI+ (C) and ESI– mode (D). IB-MALE denotes the male worms from BALB/c mice, which are marked with green plus sign. IB-FEMALE denotes the female worms from BALB/c mice, which are marked with red triangles. IS-MALE denotes the male worms from SCID mice, which are marked with blue diamonds. IS-FEMALE denotes the female worms from SCID mice, which are marked with purple cross. QC denotes the quality control samples, which are marked with pink triangles. The sample size is 5 in each group of worm samples.
FIGURE 2
Discrimination between the S. japonicum worms from SCID mice and those from BALB/c mice with S. japonicum infection for 35 days based on ESI+ and ESI- mode-derived metabolic phenotypes and heatmaps of the differential metabolites between the compared groups. (A,B): Orthogonal partial least-squares discriminant analysis (OPLS-DA) score plots in ESI+ mode (A) and ESI– mode (B) for comparison between male worms from SCID mice and those from BALB/c mice (IB-MALE denotes the male worms from BALB/c mice and IS-MALE denotes the male worms from SCID mice). (C): Heatmap of the differential metabolites between male worms from SCID mice and those from BALB/c mice. (D,E): Orthogonal partial least-squares discriminant analysis (OPLS-DA) score plots in ESI+ mode (D) and ESI– mode (E) for comparison between female worms from SCID mice and those from BALB/c mice (IB-FEMALE denotes the female worms from BALB/c mice and IS-FEMALE denotes the female worms from SCID mice). (F): Heatmap of the differential metabolites between female worms from SCID mice and those from BALB/c mice. For the heatmaps, normalized signal intensities (log2 transformed and row adjustment) are visualized as a color spectrum and the scale from least abundant to highest ranges is from –3.0 to 3.0 as shown in the colorbar. Green indicates decreased expression, whereas red indicates increased expression of the detected metabolites between compared groups. The sample size is 5 in each group of worm samples.
FIGURE 3
Scatter plots of differential metabolites with FC ≥ 1.5 or FC ≤ 0.5 in comparison groups. (A–E): Differential metabolites between IS-MALE vs. IB-MALE; (F–J): Differential metabolites between IS-FEMALE vs. IB-FEMALE. The sample size is 5 in each group of worm samples.
FIGURE 4
Comparison of metabolomics between the S. japonicum worms from SCID mice and those from BALB/c mice with S. japonicum infection for 35 days heatmaps of their differential metabolites. (A): Differential metabolites across comparison groups showing unique and common metabolites between IS-MALE vs. IB-MALE and IS-FEMALE vs. IB-FEMALE. Venn diagram displays comparatively the differentially expressed metabolites. All the differentially expressed metabolites are clustered into two comparison groups represented by two circles. The sum of all the figures in one circle represents the number of differentially expressed metabolites in one comparison group (e.g., IS-MALE vs. IB-MALE). The overlapping part of the two circles represents the number of differentially expressed metabolites shared between the two comparison groups. The single-layer part represents the number of metabolites distinctly found in a certain comparison group. (B–D): Heatmap of the common (B) and distinct [(C) is for male worms and (D) is for female worms] differential metabolites between male worms and female worms. For the heatmaps, normalized signal intensities (log2 transformed and row adjustment) are visualized as a color spectrum and the scale from least abundant to highest ranges is from –3.0 to 3.0 as shown in the colorbar. Green indicates decreased expression, whereas red indicates increased expression of the detected metabolites between compared groups. The sample size is 5 in each group of worm samples.
FIGURE 5
Enrichment analysis of the differential metabolites across comparison groups. (A): Top 8 enriched metabolite terms of the differentially expressed metabolites of IS-MALE vs. IB-MALE. The bars on _x_-axis represent the number of metabolites for the chemical classes mentioned on the _y_-axis. (B): Enriched metabolite sets of the differentially expressed metabolites of IS-MALE vs. IB-MALE. (C): Top 11 enriched metabolite terms of the differentially expressed metabolites of IS-FEMALE vs. IB-FEMALE. (D): Enriched metabolite sets of the differentially expressed metabolites of IS-FEMALE vs. IB-FEMALE. (E): Enriched metabolite terms of the common differentially expressed metabolites between IS-MALE vs. IB-MALE and IS-FEMALE vs. IB-FEMALE. (F): Enriched metabolite sets of the differentially expressed metabolites between IS-MALE vs. IB-MALE and IS-FEMALE vs. IB-FEMALE. (G): Enriched metabolite terms of the differentially expressed metabolites distinct in IS-MALE vs. IB-MALE. (H): Enriched metabolite sets of the differentially expressed metabolites distinct in IS-MALE vs. IB-MALE. (I): Enriched metabolite terms of the differentially expressed metabolites distinct in IS-FEMALE vs. IB-FEMALE. (J): Enriched metabolite sets of the differentially expressed metabolites distinct in IS-FEMALE vs. IB-FEMALE.
FIGURE 6
The metabolism of retinoids. Retinal can be originally formed by the cleavage of proretinoid carotenoids such as beta-carotene by the enzyme BCMO1 (not shown here). Retinol is formed by the reversible reduction of retinal by one of the retinal reductase family members. The enzyme lecithin:retinol acyl-transferase (LRAT) [with an ortholog gene called diacylglycerol _O_-acyltransferase (DGAT) in _Schistosoma_] synthesizes retinyl esters by transferring a fatty acyl moiety from the sn-1 position of membrane phosphatidyl choline such as lecithin [PC(22:6/20:1)] to retinol. Unesterified retinol is liberated from retinyl ester stores by the catalysis of a retinyl ester hydrolase (REH). Retinol is oxidized by catalysis of one retinol dehydrogenase (RDH) to retinal, which is then irreversibly oxidized by one retinal dehydrogenase (RALDH) to form transcriptionally active retinoic acid. Retinoic acid is finally oxidized/catabolized to more water-soluble hydroxy- and oxo- forms by one of several cytochrome P450 enzyme family members.
FIGURE 7
Transcriptional verification of some genes involved in retinol metabolism and the associated meiosis process by qPCR. The tested four genes in “retinol metabolism” are: (A) S. japonicum diacylglycerol _O_-acyltransferase 1 (SjDGAT1), (B) short chain dehydrogenase/reductase (SjDHRS), (C) aldehyde dehydrogenase 1B1 precursor (SjALDH1B1), and (D) retinol dehydrogenase 12 (SjRDH12). The tested eight genes involved in meiosis are: (E) S. japonicum meiotic recombination protein SPO11 (SjSPO11), (F) double-strand break repair protein MRE11A (SjMRE11), (G) meiotic nuclear division protein 1-like protein (SjMND1), (H) S-phase kinase-associated protein 1A (Sjskp1a), (I) polo-like kinase 1 (SjPLK1), (J) polo-like kinase 4 (SjPLK4), (K) DNA repair protein RAD51 (SjRAD51), and (L) meiotic recombinase DMC1 (SjDMC1). S_F: IS-FEMALE, B_F: IB-FEMALE, S_M: IS-MALE, and B_M: IB-MALE. Two duplicate examinations were performed for each group of worm samples.
References
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