Human Milk Extracellular Vesicles Modulate Inflammation and Cell Survival in Intestinal and Immune Cells

Overview

Journal Pediatr Res

Specialties Biology
Pediatrics

Date 2024 Nov 28

PMID 39609615

Authors

Brett Vahkal

Illimar Altosaar

Ardeshir Ariana

Josie Jabbour

Falia Pantieras

Redaet Daniel

Eric Tremblay

Subash Sad

Jean-Francois Beaulieu

Marceline Cote

Emanuela Ferretti

Affiliations

Soon will be listed here.

Abstract

Human milk contains extracellular vesicles (EVs) that carry bioactive molecules such as microRNA, to the newborn intestine. The downstream effects of EV cargo on signaling and immune modulation may shield neonates against inflammatory diseases, including necrotizing enterocolitis. Premature infants are especially at risk, while human milk-feeding may offer protection. The effect of gestational-age specific term and preterm EVs from transitional human milk was characterized on human intestinal epithelial cells (HIECs and Caco-2), primary macrophages, and THP-1 monocytes. We hypothesized that term and preterm EVs differentially influence immune-related cytokines and cell death. We found that preterm EVs were enriched in CD14 surface marker, while both term and preterm EVs increased epidermal growth factor secretion. Following inflammatory stimuli, only term EVs inhibited secretion of IL-6 in HIECs, and reduced expression of pro-inflammatory cytokine IL-1β in macrophages. Term and preterm EVs inhibited secretion of IL-1β and reduced inflammasome related cell death. We proposed that human milk EVs regulate immune-related signaling via their conserved microRNA cargo, which could promote tolerance and a homeostatic immune response. These findings provide basis for further studies into potential therapeutic supplementation with EVs in vulnerable newborn populations by considering functional, gestational age-specific effects. IMPACT: This study reveals distinct functional differences between term and preterm transitional human milk extracellular vesicles (EVs) highlighting the importance of gestational age in their bioactivity. Term EVs uniquely inhibited IL-6 secretion, IL-1β expression, and apoptosis following inflammatory stimuli. Both term and preterm human milk EVs reduced IL-1β secretion and inflammasome-induced cell death. Conserved human milk extracellular vesicle microRNA cargo could be a mediator of the anti-inflammatory effects, particularly targeting cytokine production, the inflammasome, and programmed cell death. These findings underscore the importance of considering gestational age in future research exploring the therapeutic potential of human milk extracellular vesicles to prevent or treat intestinal inflammatory diseases in neonates.

References

Sullivan S, Schanler R, Kim J, Patel A, Trawoger R, Kiechl-Kohlendorfer U . An exclusively human milk-based diet is associated with a lower rate of necrotizing enterocolitis than a diet of human milk and bovine milk-based products. J Pediatr. 2009; 156(4):562-7.e1. DOI: 10.1016/j.jpeds.2009.10.040. View

Zonneveld M, van Herwijnen M, Fernandez-Gutierrez M, Giovanazzi A, Marit de Groot A, Kleinjan M . Human milk extracellular vesicles target nodes in interconnected signalling pathways that enhance oral epithelial barrier function and dampen immune responses. J Extracell Vesicles. 2021; 10(5):e12071. PMC: 7944547. DOI: 10.1002/jev2.12071. View

Tong L, Zhang S, Liu Q, Huang C, Hao H, Tan M . Milk-derived extracellular vesicles protect intestinal barrier integrity in the gut-liver axis. Sci Adv. 2023; 9(15):eade5041. PMC: 10096581. DOI: 10.1126/sciadv.ade5041. View

Izumi H, Tsuda M, Sato Y, Kosaka N, Ochiya T, Iwamoto H . Bovine milk exosomes contain microRNA and mRNA and are taken up by human macrophages. J Dairy Sci. 2015; 98(5):2920-33. DOI: 10.3168/jds.2014-9076. View

Swieboda D, Johnson E, Beaver J, Haddad L, Enninga E, Hathcock M . Baby's First Macrophage: Temporal Regulation of Hofbauer Cell Phenotype Influences Ligand-Mediated Innate Immune Responses across Gestation. J Immunol. 2020; 204(9):2380-2391. PMC: 7870092. DOI: 10.4049/jimmunol.1901185. View

Torow N, Marsland B, Hornef M, Gollwitzer E . Neonatal mucosal immunology. Mucosal Immunol. 2016; 10(1):5-17. DOI: 10.1038/mi.2016.81. View

Cho S, Berger P, Nold-Petry C, Nold M . The immunological landscape in necrotising enterocolitis. Expert Rev Mol Med. 2016; 18:e12. PMC: 5001507. DOI: 10.1017/erm.2016.13. View

Nino D, Sodhi C, Hackam D . Necrotizing enterocolitis: new insights into pathogenesis and mechanisms. Nat Rev Gastroenterol Hepatol. 2016; 13(10):590-600. PMC: 5124124. DOI: 10.1038/nrgastro.2016.119. View

Dreschers S, Ohl K, Schulte N, Tenbrock K, Orlikowsky T . Impaired functional capacity of polarised neonatal macrophages. Sci Rep. 2020; 10(1):624. PMC: 6968972. DOI: 10.1038/s41598-019-56928-4. View

10.

Zhu F, Wang L, Gong Z, Wang Y, Gao Y, Cai W . Blockage of NLRP3 inflammasome activation ameliorates acute inflammatory injury and long-term cognitive impairment induced by necrotizing enterocolitis in mice. J Neuroinflammation. 2021; 18(1):66. PMC: 7937302. DOI: 10.1186/s12974-021-02111-4. View

11.

Spong C, Mercer B, DAlton M, Kilpatrick S, Blackwell S, Saade G . Timing of indicated late-preterm and early-term birth. Obstet Gynecol. 2011; 118(2 Pt 1):323-333. PMC: 3160133. DOI: 10.1097/AOG.0b013e3182255999. View

12.

Dvorak B, Fituch C, Williams C, Hurst N, Schanler R . Increased epidermal growth factor levels in human milk of mothers with extremely premature infants. Pediatr Res. 2003; 54(1):15-9. DOI: 10.1203/01.PDR.0000065729.74325.71. View

13.

Plaza-Zamora J, Sabater-Molina M, Rodriguez-Palmero M, Rivero M, Bosch V, Nadal J . Polyamines in human breast milk for preterm and term infants. Br J Nutr. 2013; 110(3):524-8. DOI: 10.1017/S0007114512005284. View

14.

Vahkal B, Altosaar I, Tremblay E, Gagne D, Huttman N, Minic Z . Gestational age at birth influences protein and RNA content in human milk extracellular vesicles. J Extracell Biol. 2024; 3(1):e128. PMC: 11080785. DOI: 10.1002/jex2.128. View

15.

Freiria-Martinez L, Iglesias-Martinez-Almeida M, Rodriguez-Jamardo C, Rivera-Baltanas T, Comis-Tuche M, Rodrigues-Amorim D . Proteomic analysis of exosomes derived from human mature milk and colostrum of mothers with term, late preterm, or very preterm delivery. Anal Methods. 2023; 15(37):4905-4917. DOI: 10.1039/d3ay01114c. View

16.

Admyre C, Johansson S, Qazi K, Filen J, Lahesmaa R, Norman M . Exosomes with immune modulatory features are present in human breast milk. J Immunol. 2007; 179(3):1969-78. DOI: 10.4049/jimmunol.179.3.1969. View

17.

de la Torre Gomez C, Goreham R, Bech Serra J, Nann T, Kussmann M . "Exosomics"-A Review of Biophysics, Biology and Biochemistry of Exosomes With a Focus on Human Breast Milk. Front Genet. 2018; 9:92. PMC: 5881086. DOI: 10.3389/fgene.2018.00092. View

18.

Welsh J, Killingsworth B, Kepley J, Traynor T, Cook S, Savage J . MPA software enables stitched multiplex, multidimensional EV repertoire analysis and a standard framework for reporting bead-based assays. Cell Rep Methods. 2022; 2(1):100136. PMC: 9017130. DOI: 10.1016/j.crmeth.2021.100136. View

19.

Pageot L, Perreault N, Basora N, Francoeur C, Magny P, Beaulieu J . Human cell models to study small intestinal functions: recapitulation of the crypt-villus axis. Microsc Res Tech. 2000; 49(4):394-406. DOI: 10.1002/(SICI)1097-0029(20000515)49:4<394::AID-JEMT8>3.0.CO;2-K. View

20.

Vachon P, Beaulieu J . Transient mosaic patterns of morphological and functional differentiation in the Caco-2 cell line. Gastroenterology. 1992; 103(2):414-23. DOI: 10.1016/0016-5085(92)90829-n. View