LncRNA and mRNA profiles of human milk-derived exosomes and their possible roles in protecting against necrotizing enterocolitis

Yan, Xiangyun; Liu, Linjie; Yao, Shuwen; Chen, Yanjie; Yu, Qinlei; Jiang, Chengyao; Chen, Wenjuan; Chen, Xiaohui; Han, Song

doi:10.1039/d2fo01866g

Cited by 7 publications

(7 citation statements)

References 52 publications

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“…Protein complexes can also mimic the appearance and size of EVs in NTA or RPS. To circumvent these pitfalls, it is common to perform HMEV isolations on large starting volumes, which allow for multiple processing steps to enrich for EVs and remove other bioactive factors that may complicate data interpretation (Chen et al., 2021 ; Miyake et al., 2020 ; Mukhopadhya et al., 2021 ; Tong et al., 2023 ; Yan et al., 2022 ; Zonneveld et al., 2021 ). This can be prohibitive if sample volumes are limited (≤2 mL).…”

Section: Discussionmentioning

confidence: 99%

Neonatal enteroids absorb extracellular vesicles from human milk‐fed infant digestive fluid

Yung,

Zhang,

Kuhn

et al. 2024

J of Extracellular Vesicle

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Human milk contains extracellular vesicles (HMEVs). Pre‐clinical models suggest that HMEVs may enhance intestinal function and limit inflammation; however, it is unknown if HMEVs or their cargo survive neonatal human digestion. This limits the ability to leverage HMEV cargo as additives to infant nutrition or as therapeutics. This study aimed to develop an EV isolation pipeline from small volumes of human milk and neonatal intestinal contents after milk feeding (digesta) to address the hypothesis that HMEVs survive in vivo neonatal digestion to be taken up intestinal epithelial cells (IECs). Digesta was collected from nasoduodenal sampling tubes or ostomies. EVs were isolated from raw and pasteurized human milk and digesta by density‐gradient ultracentrifugation following two‐step skimming, acid precipitation of caseins, and multi‐step filtration. EVs were validated by electron microscopy, western blotting, nanoparticle tracking analysis, resistive pulse sensing, and super‐resolution microscopy. EV uptake was tested in human neonatal enteroids. HMEVs and digesta EVs (dEVs) show typical EV morphology and are enriched in CD81 and CD9, but depleted of β‐casein and lactalbumin. HMEV and some dEV fractions contain mammary gland‐derived protein BTN1A1. Neonatal human enteroids rapidly take up dEVs in part via clathrin‐mediated endocytosis. Our data suggest that EVs can be isolated from digestive fluid and that these dEVs can be absorbed by IECs.

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Section: Discussionmentioning

confidence: 99%

Neonatal enteroids absorb extracellular vesicles from human milk‐fed infant digestive fluid

Yung,

Zhang,

Kuhn

et al. 2024

J of Extracellular Vesicle

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“…Protein complexes can also mimic the appearance and size of EVs in NTA or RPS. To circumvent these pitfalls, it is common to perform HMEV isolations on large starting volumes, which allow for multiple processing steps to enrich for EVs and remove other bioactive factors that may complicate data interpretation (15, 17, 18, 46, 79, 80). This can be prohibitive if sample volumes are limited (≤ 2mL).…”

Section: Discussionmentioning

confidence: 99%

Neonatal enteroids absorb extracellular vesicles from human milk-fed infant digestive fluid

Yung,

Zhang,

Kuhn

et al. 2023

Preprint

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Human milk contains extracellular vesicles (HMEVs). Pre-clinical models suggest that HMEVs may enhance intestinal function and limit inflammation; however, it is unknown if HMEVs or their cargo survive neonatal human digestion. This limits the ability to leverage HMEV cargo as additives to infant nutrition or as therapeutics. This study aimed to develop an EV isolation pipeline from small volumes of human milk and neonatal intestinal contents after milk feeding (digesta) to address the hypothesis that HMEVs survive in vivo neonatal digestion to be taken up intestinal epithelial cells (IECs). Digesta was collected from nasoduodenal sampling tubes. EVs were isolated from raw and pasteurized human milk and digesta by density-gradient ultracentrifugation following two-step skimming, acid precipitation of caseins, and multi-step filtration. EVs were validated by electron microscopy, nanoparticle tracking analysis, and western blotting. EV uptake and effects were tested in human neonatal enteroids. HMEVs and digesta EVs (dHMEVs) show typical EV morphology and are enriched in CD81 and CD9, but depleted of β-casein and lactalbumin. HMEVs and dHMEVs contain mammary gland-derived protein BTN1A1. Neonatal human enteroids rapidly take up dHMEVs in part via clathrin-mediated endocytosis. Our data suggest that HMEVs survive to the duodenum and can be absorbed by IECs.

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“…However, differentially expressed mRNA were not functionally characterised by term of birth, but rather described in the context of interactions with long‐coding RNAs. These specific interactions focused on upregulated preterm EV mRNAs, which were determined to be enriched in immune, signalling and metabolic pathways (Yan et al., 2022).…”

Section: Introductionmentioning

confidence: 99%

“…Though differential composition of HM EV cargo of proteins has been described following both term and preterm birth (Chen et al., 2021; Kahn et al., 2018; Wang et al., 2019; Yan et al., 2022), an overall limited number of individual proteins with quantitative information have been identified. Additional low abundance proteins remain elusive, but their identification could expand functional interpretations for HM EVs.…”

Section: Introductionmentioning

confidence: 99%

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Gestational age at birth influences protein and RNA content in human milk extracellular vesicles

Vahkal,

Altosaar,

Tremblay

et al. 2023

J of Extracellular Bio

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Human milk extracellular vesicles (HM EVs) are proposed to protect against disease development in infants. This protection could in part be facilitated by the bioactive EV cargo of proteins and RNA. Notably, mothers birth infants of different gestational ages with unique needs, wherein the EV cargo of HM may diverge. We collected HM from lactating mothers within two weeks of a term or preterm birth. Following purification of EVs, proteins and mRNA were extracted for proteomics and sequencing analyses, respectively. Over 2000 protein groups were identified, and over 8000 genes were quantified. The total number of proteins and mRNA did not differ significantly between the two conditions, while functional bioinformatics of differentially expressed cargo indicated enrichment in immunoregulatory cargo for preterm HM EVs. In term HM EVs, significantly upregulated cargo was enriched in metabolism‐related functions. Based on gene expression signatures from HM‐contained single cell sequencing data, we proposed that a larger portion of preterm HM EVs are secreted by immune cells, whereas term HM EVs contain more signatures of lactocyte epithelial cells. Proposed differences in EV cargo could indicate variation in mother's milk based on infants’ gestational age and provide basis for further functional characterisation.

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LncRNA and mRNA profiles of human milk-derived exosomes and their possible roles in protecting against necrotizing enterocolitis

Abstract: Necrotizing enterocolitis (NEC) is one of the most commonly severe diseases afflicting premature infants. Our previous studies suggests that human milk-derived exosomes (HM-Exos) have a potential therapeutic effect on NEC....

Cited by 7 publications

References 52 publications

Neonatal enteroids absorb extracellular vesicles from human milk‐fed infant digestive fluid

Neonatal enteroids absorb extracellular vesicles from human milk‐fed infant digestive fluid

Neonatal enteroids absorb extracellular vesicles from human milk-fed infant digestive fluid

Gestational age at birth influences protein and RNA content in human milk extracellular vesicles

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