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J Dairy Sci. 2020 Jan;103(1):16-29. doi: 10.3168/jds.2019-16880. Epub 2019 Oct 31.

Complexity of the microRNA transcriptome of cow milk and milk-derived extracellular vesicles isolated via differential ultracentrifugation.

Author information

1
Centre Hospitalier Universitaire de Québec Research Center/Centre Hospitalier de l'Université Laval, Quebec G1V 4G2, Canada; Department of Microbiology, Infectious Diseases and Immunology and Faculty of Medicine, Université Laval, Quebec G1V 0A6, Canada.
2
Centre Hospitalier Universitaire de Québec Research Center/Centre Hospitalier de l'Université Laval, Quebec G1V 4G2, Canada; Department of Molecular Medicine, Faculty of Medicine, Université Laval, Axe Endocrinologie - Néphrologie du Centre de recherche du Centre Hospitalier Universitaire de Québec - Université Laval, Québec City, Québec, Canada.
3
Centre Hospitalier Universitaire de Québec Research Center/Centre Hospitalier de l'Université Laval, Quebec G1V 4G2, Canada; Department of Microbiology, Infectious Diseases and Immunology and Faculty of Medicine, Université Laval, Quebec G1V 0A6, Canada. Electronic address: patrick.provost@crchudequebec.ulaval.ca.

Abstract

MicroRNAs (miRNAs) are small gene-regulatory noncoding RNA that are highly enriched in cow milk. They are encapsulated in different extracellular vesicle (EV) subsets that protect them from the extracellular milieu and the harsh conditions of the gastrointestinal tract during digestion. Here, we isolated pellets enriched in 4 different EV subsets, via differential ultracentrifugation of commercial cow milk: 12,000 × g (P12K), 35,000 × g (P35K), 70,000 × g (P70K), and 100,000 × g (P100K). Small RNA sequencing (sRNA-Seq) analyses revealed an unprecedented level of diversity in the complete miRNA repertoire and features of unfractionated cow milk and derived EV subsets. Although 5 miRNA sequences represented more than 50% of all miRNAs, milk EV exhibited heterogeneous content of miRNAs and isomeric variants (termed isomiR): P100K EV were enriched in reference miRNA sequences, and P12K and P35K EV in related isomiR. Incubation of milk EV with human cultured HeLa cells led to cellular enrichment in miRNA miR-223, which was concomitant with decreased expression of a reporter gene placed under the control of miR-223, thereby demonstrating the functionality of miR-223. These results suggest that cow milk EV may transfer their miRNAs to human cells and regulate recipient cell gene expression programming in a manner as complex as that of their miRNA transcriptome. The biological activity and relevance of the different milk EV subsets and bioactive mediators, including small noncoding RNA, in health and disease, warrants further investigation.

KEYWORDS:

bovine; exosome; extracellular vesicle; microRNA; small RNA

PMID:
31677838
DOI:
10.3168/jds.2019-16880
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