Metabolome alterations in a mouse model support microbiome-metabolite interactions in a cohort of children with cow's milk allergy
- Author
- Ellen De Paepe (UGent) , Vera Plekhova (UGent) , Lars Vereecke (UGent) , Myriam Van Winckel (UGent) , Marilyn De Graeve (UGent) , Tom Van de Wiele (UGent) , Eric Cox (UGent) and Lynn Vanhaecke (UGent)
- Organization
- Abstract
- Food allergies are a growing epidemic and constitute a major threat to our well-being. Cow’s milk allergy is one of the first allergies to arise to various proteins, mainly beta-lactoglobulin (BLG) and casein. In this study, we used two different protocols to sensitize two mouse strains to BLG. Murine fecal metabolomics was corroborated by microbiome-metabolome findings in a cohort of allergic children and revealed 275 significant metabolite alterations and 5 impaired pathways in food allergy. Several of these pathways, i.e. bile acid, tryptophan and energy metabolism have been associated with microbial dysbiosis, as confirmed in our cohort of allergic children.
- In Westernized societies, food allergies (FAs) are a growing epidemic and are considered a major cost to our health care system. Cow’s milk allergy (CMA) is one of the first allergies to arise in early childhood and may occur to various milk proteins or allergens, of which beta-lactoglobulin (BLG) and casein are the most important. In this study, we used two different protocols (cholera toxin (CT) or an oil emulsion) to sensitize two mouse strains (Balb/C and C57Bl/6; n=6 per group; 120 in total) to BLG and controls. After sensitization, mice were challenged orally, and their clinical signs monitored. Plasma and spleen cells were collected for antibody (IgE and IgG1) and cytokine (IL-10 and IFN-γ) determination, fecal samples were subjected to metabolomics using UHPLC-Q-Orbitrap-HRMS[1]. Spearman correlations between blood IgE levels and normalized fecal metabolite levels were calculated. Jejunum and ileum were collected for histological evaluation after HE-staining. These results were further supported by fecal microbiome-metabolome data from our population of allergic (IgE mediated-CMA (IgE-CMA); n=24) and healthy (n=23) children, on which polar metabolomics[1], lipidomics[2] and 16S rRNA metasequencing[3] were performed. For both the murine experiment and the patient cohort, univariate and multivariate statistical analysis, as well as pathway analysis, were performed, followed by MS/MS analysis for annotation of highly correlating or discriminative metabolites. Anaphylaxis was observed in Balb/C mice receiving BLG and CT, next to elevated antibody -and histamine levels, and lower anti-inflammatory cytokine IFN-γ levels. Moderate to severe intestinal abnormality in histology was discovered in the control group receiving CT and the allergic group receiving CT and BLG, respectively. Orthogonal Partial Least-Squares Discriminant Analysis (OPLS-DA) confirmed clinical findings, showing valid Q2 (>0.7) and R2(Y) (>0.99), significant p-values (<0.05), and good permutation testing (n=100). Univariate statistical analysis revealed increased levels of 3-hydroxykynurerin, sucrose, hippuric acid and glycerophosphorylcholine. Tryptophan (p=0.044) and starch and sucrose (p=0.048) were most significantly affected. In allergic children, good OPLS-DA modeling was observed, while univariate statistical analysis on annotated metabolites confirmed significant alterations in the tryptophan, purine and energy pathways, while functional analysis on the untargeted dataset revealed significant impairment of the tryptophan (p=0.042) and bile acid (p=0.038) metabolism, in addition to several pathways linked to energy metabolism. The observed alterations can be mainly attributed to microbial dysbiosis, which was confirmed by a decreased alpha and beta diversity in children with CMA (p=0.030 and 0.034, respectively). It is hypothesized that this dysbiosis and its concomitant metabolic disturbances are a direct consequence of sensitization and as such precede CMA mediated inflammation, as no metabolic signs of dysbiosis were observed in control mice. Clinical signs of discomfort, but not anaphylaxis, were observed in the C57Bl/6 mice receiving BLG in oil, which was associated with borderline significant (p-value < 0.1) increased levels of IgG1. As such, this model could represent a more accurate model for non-IgE mediated CMA in children. Our results suggest that metabolic alterations in Balb/C mice sensitized with BLG and CT in PBS optimally resemble IgE-CMA in children. Furthermore, we hypothesize that sensitization is the true causal factor of microbial dysbiosis in food allergies, as no metabolic signs of dysbiosis were observed in control mice and metabolic markers of allergy emerge at a slower pace than dysbiosis markers. More in-depth data exploration of the data through multi-omics, to map microbiome-metabolome-lipidome interactions, is ongoing.
Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-01GW21Q0TJ3DW4HSZGY4YWF2GH
- MLA
- De Paepe, Ellen, et al. “Metabolome Alterations in a Mouse Model Support Microbiome-Metabolite Interactions in a Cohort of Children with Cow’s Milk Allergy.” Benelux Metabolomics Days, Abstracts, 2022.
- APA
- De Paepe, E., Plekhova, V., Vereecke, L., Van Winckel, M., De Graeve, M., Van de Wiele, T., … Vanhaecke, L. (2022). Metabolome alterations in a mouse model support microbiome-metabolite interactions in a cohort of children with cow’s milk allergy. Benelux Metabolomics Days, Abstracts. Presented at the Benelux Metabolomics Days 2022, Utrecht, The Netherlands.
- Chicago author-date
- De Paepe, Ellen, Vera Plekhova, Lars Vereecke, Myriam Van Winckel, Marilyn De Graeve, Tom Van de Wiele, Eric Cox, and Lynn Vanhaecke. 2022. “Metabolome Alterations in a Mouse Model Support Microbiome-Metabolite Interactions in a Cohort of Children with Cow’s Milk Allergy.” In Benelux Metabolomics Days, Abstracts.
- Chicago author-date (all authors)
- De Paepe, Ellen, Vera Plekhova, Lars Vereecke, Myriam Van Winckel, Marilyn De Graeve, Tom Van de Wiele, Eric Cox, and Lynn Vanhaecke. 2022. “Metabolome Alterations in a Mouse Model Support Microbiome-Metabolite Interactions in a Cohort of Children with Cow’s Milk Allergy.” In Benelux Metabolomics Days, Abstracts.
- Vancouver
- 1.De Paepe E, Plekhova V, Vereecke L, Van Winckel M, De Graeve M, Van de Wiele T, et al. Metabolome alterations in a mouse model support microbiome-metabolite interactions in a cohort of children with cow’s milk allergy. In: Benelux Metabolomics Days, Abstracts. 2022.
- IEEE
- [1]E. De Paepe et al., “Metabolome alterations in a mouse model support microbiome-metabolite interactions in a cohort of children with cow’s milk allergy,” in Benelux Metabolomics Days, Abstracts, Utrecht, The Netherlands, 2022.
@inproceedings{01GW21Q0TJ3DW4HSZGY4YWF2GH,
abstract = {{Food allergies are a growing epidemic and constitute a major threat to our well-being. Cow’s milk allergy is one of the first allergies to arise to various proteins, mainly beta-lactoglobulin (BLG) and casein. In this study, we used two different protocols to sensitize two mouse strains to BLG. Murine fecal metabolomics was corroborated by microbiome-metabolome findings in a cohort of allergic children and revealed 275 significant metabolite alterations and 5 impaired pathways in food allergy. Several of these pathways, i.e. bile acid, tryptophan and energy metabolism have been associated with microbial dysbiosis, as confirmed in our cohort of allergic children.}},
author = {{De Paepe, Ellen and Plekhova, Vera and Vereecke, Lars and Van Winckel, Myriam and De Graeve, Marilyn and Van de Wiele, Tom and Cox, Eric and Vanhaecke, Lynn}},
booktitle = {{Benelux Metabolomics Days, Abstracts}},
language = {{eng}},
location = {{Utrecht, The Netherlands}},
title = {{Metabolome alterations in a mouse model support microbiome-metabolite interactions in a cohort of children with cow's milk allergy}},
year = {{2022}},
}