TY - JOUR
T1 - Dietary bovine milk miRNAs transported in extracellular vesicles are partially stable during GI digestion, are bioavailable and reach target tissues but need a minimum dose to impact on gene expression
AU - López de Las Hazas, María Carmen
AU - Del Pozo-Acebo, Lorena
AU - Hansen, Maria S.
AU - Gil-Zamorano, Judit
AU - Mantilla-Escalante, Diana C.
AU - Gómez-Coronado, Diego
AU - Marín, Francisco
AU - Garcia-Ruiz, Almudena
AU - Rasmussen, Jan T.
AU - Dávalos, Alberto
N1 - Publisher Copyright:
© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany.
PY - 2022/3/1
Y1 - 2022/3/1
N2 - PURPOSE: Extracellular RNAs are unstable and rapidly degraded unless protected. Bovine-milk extracellular vesicles (EVs) confer protection to dietary miRNAs, although it remains unclear whether this importantly improves their chances of reaching host target cells to exert biological effects. METHODS: Caco-2, HT-29, Hep-G2 and FHs-74 cell lines were exposed to natural/labelled milk EVs to evaluate cellular uptake. Five frequently reported human milk miRNAs (miR-146b-5p, miR-148a-3p, miR-30a-5p, miR-26a-5p, and miR-22-3p) were loaded into EVs. The intracellular concentration of each miRNA in cells was determined. In addition, an animal study giving an oral dose of loaded EVs in C57BL6/ mice were performed. Gene expression regulation was assessed by microarray analysis. RESULTS: Digestive stability analysis showed high overall degradation of exogenous miRNAs, although EV-protected miRNAs better resisted gastrointestinal digestion compared to free miRNAs (tenfold higher levels). Importantly, orally delivered EV-loaded miRNAs reached host organs, including brain, in mice. However, no biological effect has been identified. CONCLUSION: Milk EVs protect miRNAs from degradation and facilitate cellular uptake. miRNA concentration in EVs from bovine milk might be insufficient to produce gene modulation. Nevertheless, sizable amounts of exogenous miRNAs may be loaded into EVs, and orally delivered EV-loaded miRNAs can reach tissues in vivo, increasing the possibility of exerting biological effects. Further investigation is justified as this could have an impact in the field of nutrition and health (i.e., infant formulas elaboration).
AB - PURPOSE: Extracellular RNAs are unstable and rapidly degraded unless protected. Bovine-milk extracellular vesicles (EVs) confer protection to dietary miRNAs, although it remains unclear whether this importantly improves their chances of reaching host target cells to exert biological effects. METHODS: Caco-2, HT-29, Hep-G2 and FHs-74 cell lines were exposed to natural/labelled milk EVs to evaluate cellular uptake. Five frequently reported human milk miRNAs (miR-146b-5p, miR-148a-3p, miR-30a-5p, miR-26a-5p, and miR-22-3p) were loaded into EVs. The intracellular concentration of each miRNA in cells was determined. In addition, an animal study giving an oral dose of loaded EVs in C57BL6/ mice were performed. Gene expression regulation was assessed by microarray analysis. RESULTS: Digestive stability analysis showed high overall degradation of exogenous miRNAs, although EV-protected miRNAs better resisted gastrointestinal digestion compared to free miRNAs (tenfold higher levels). Importantly, orally delivered EV-loaded miRNAs reached host organs, including brain, in mice. However, no biological effect has been identified. CONCLUSION: Milk EVs protect miRNAs from degradation and facilitate cellular uptake. miRNA concentration in EVs from bovine milk might be insufficient to produce gene modulation. Nevertheless, sizable amounts of exogenous miRNAs may be loaded into EVs, and orally delivered EV-loaded miRNAs can reach tissues in vivo, increasing the possibility of exerting biological effects. Further investigation is justified as this could have an impact in the field of nutrition and health (i.e., infant formulas elaboration).
KW - Exosomes
KW - Extracellular vesicles
KW - In vitro digestion
KW - Milk miRNAs
KW - Transcriptome
KW - Transfection
KW - Caco-2 Cells
KW - MicroRNAs/genetics
KW - Gene Expression
KW - Humans
KW - Digestion
KW - Extracellular Vesicles/metabolism
KW - Animals
KW - Milk, Human/metabolism
KW - Mice
UR - http://www.scopus.com/inward/record.url?scp=85118305799&partnerID=8YFLogxK
U2 - 10.1007/s00394-021-02720-y
DO - 10.1007/s00394-021-02720-y
M3 - Journal article
C2 - 34716465
AN - SCOPUS:85118305799
SN - 1436-6207
VL - 61
SP - 1043
EP - 1056
JO - European Journal of Nutrition
JF - European Journal of Nutrition
IS - 2
ER -