The human milk oligosaccharide 3′sialyllactose reduces low-grade inflammation and atherosclerosis development in mice
Ariane R. Pessentheiner, Nathanael J. Spann, Chloe A. Autran, Tae Gyu Oh, Kaare V. Grunddal, Joanna K.C. Coker, Chelsea D. Painter, Bastian Ramms, Austin W.T. Chiang, Chen-Yi Wang, Jason Hsiao, Yiwen Wang, Anthony Quach, Laela M. Booshehri, Alexandra Hammond, Chiara Tognaccini, Joanna Latasiewicz, Lisa Willemsen, Karsten Zengler, Menno P.J. de Winther, Hal M. Hoffman, Martin Philpott, Adam P. Cribbs, Udo Oppermann, Nathan E. Lewis, Joseph L. Witztum, Ruth Yu, Annette R. Atkins, Michael Downes, Ron M. Evans, Christopher K. Glass, Lars Bode, Philip L.S.M. Gordts
Ariane R. Pessentheiner, Nathanael J. Spann, Chloe A. Autran, Tae Gyu Oh, Kaare V. Grunddal, Joanna K.C. Coker, Chelsea D. Painter, Bastian Ramms, Austin W.T. Chiang, Chen-Yi Wang, Jason Hsiao, Yiwen Wang, Anthony Quach, Laela M. Booshehri, Alexandra Hammond, Chiara Tognaccini, Joanna Latasiewicz, Lisa Willemsen, Karsten Zengler, Menno P.J. de Winther, Hal M. Hoffman, Martin Philpott, Adam P. Cribbs, Udo Oppermann, Nathan E. Lewis, Joseph L. Witztum, Ruth Yu, Annette R. Atkins, Michael Downes, Ron M. Evans, Christopher K. Glass, Lars Bode, Philip L.S.M. Gordts
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Research Article Cell biology Inflammation

The human milk oligosaccharide 3′sialyllactose reduces low-grade inflammation and atherosclerosis development in mice

Abstract

Macrophages contribute to the induction and resolution of inflammation and play a central role in chronic low-grade inflammation in cardiovascular diseases caused by atherosclerosis. Human milk oligosaccharides (HMOs) are complex unconjugated glycans unique to human milk that benefit infant health and act as innate immune modulators. Here, we identify the HMO 3′sialyllactose (3′SL) as a natural inhibitor of TLR4-induced low-grade inflammation in macrophages and endothelium. Transcriptome analysis in macrophages revealed that 3′SL attenuates mRNA levels of a selected set of inflammatory genes and promotes the activity of liver X receptor (LXR) and sterol regulatory element binding protein-1 (SREBP1). These acute antiinflammatory effects of 3′SL were associated with reduced histone H3K27 acetylation at a subset of LPS-inducible enhancers distinguished by preferential enrichment for CCCTC-binding factor (CTCF), IFN regulatory factor 2 (IRF2), B cell lymphoma 6 (BCL6), and other transcription factor recognition motifs. In a murine atherosclerosis model, both s.c. and oral administration of 3′SL significantly reduced atherosclerosis development and the associated inflammation. This study provides evidence that 3′SL attenuates inflammation by a transcriptional mechanism to reduce atherosclerosis development in the context of cardiovascular disease.

Authors

Ariane R. Pessentheiner, Nathanael J. Spann, Chloe A. Autran, Tae Gyu Oh, Kaare V. Grunddal, Joanna K.C. Coker, Chelsea D. Painter, Bastian Ramms, Austin W.T. Chiang, Chen-Yi Wang, Jason Hsiao, Yiwen Wang, Anthony Quach, Laela M. Booshehri, Alexandra Hammond, Chiara Tognaccini, Joanna Latasiewicz, Lisa Willemsen, Karsten Zengler, Menno P.J. de Winther, Hal M. Hoffman, Martin Philpott, Adam P. Cribbs, Udo Oppermann, Nathan E. Lewis, Joseph L. Witztum, Ruth Yu, Annette R. Atkins, Michael Downes, Ron M. Evans, Christopher K. Glass, Lars Bode, Philip L.S.M. Gordts

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Figure 8

3′SL downregulates inflammatory pathways and upregulates cholesterol biosynthesis in atherosclerotic lesion macrophages.

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3′SL downregulates inflammatory pathways and upregulates cholesterol bio...
(A and B) Heatmaps representing Z-normalized row mRNA levels of each gene for RNA-Seq from independent biological duplicates showing the 50 most upregulated (right) or downregulated (left) genes in BMDMs at 6 hours after Pam3CSK4 ± 3′SL stimulation. (C and D) Enriched pathways identified by metascape analysis software. All significantly, differentially expressed mRNA levels of genes in the Pam3CSK4 versus Pam3CSK4 + 3′SL RNA-Seq datasets were used for the analysis with IPA (cut-off > 1.5-fold) with Z score ≥ ±2. (E) Annotated UMAP visualizations of the identified clusters of snRNA-Seq data from cells of the atherosclerotic aortic arch of water treated or 3′SL-treated male mice. Ldlr–/– male mice were fed a Western-type diet (WTD) for 10 weeks. After 2 weeks, mice were treated twice daily with oral gavage of 90 mg 3′SL in water per mouse for 6 weeks. Water gavage served as a control. (F) Relative distribution of atherosclerotic lesion macrophage subsets in water versus 3′SL-treated mice. (G) The number of significantly differentially expressed mRNA gene transcripts in macrophage subsets and their directionality upon 3′SL administration (upregulation is > 2-fold; downregulation is <-2-fold; compared with control treatment). (H) Enriched pathways identified by metascape analysis software of differentially expressed mRNA in macrophage subsets upon 3′SL administration (> 1.75-fold or < –1.75 change; compared with control treatment). (I and J) The expression of numerous genes from snRNA-Seq was significantly increased (I) or decreased (J) by 3′SL treatment in plaque macrophage subsets compared with WT. Unpaired 2-tailed Student’s t test. *P < 0.05 versus control. Data represent mean ± SEM.