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Specific milk composition in miR-30b-transgenic mice associated to offspring early gut maturation with lasting consequences on their growth

ABSTRACT: Milk composition is complex and includes numerous components essential for the offspring growth and development. Besides a high abundance of the miR-30b microRNA, milk produced by the transgenic mouse model of miR-30b-mammary deregulation displays significant changes in its fatty acid profils. Moreover, wild-type adopted pups fed with this milk present an early growth defect. Therefore, the consequences of miR-30b milk feeding on neonate gut development, a prime target of suckled milk, were investigated, along with further characterization of changes in milk composition. A broad characterization of the duodenum of wild-type pups fed with miR-30b milk was performed, using histological, transcriptomic, proteomic and intestinal permeability analyses. Milk of miR-30b foster dams was extensively analyzed using proteomic, metabolomic and lipidomic approaches and hormonal immunoassays. Pups fed with miR-30b milk showed a maturation of their gut tissue, presenting an earlier reduction in paracellular and transcellular permeability at postnatal day 5. MiR-30b milk displayed significant changes in its total lipid content, ceramides and sphingomyelin concentrations, an overabundance in nine proteins and an increase in insulin and leptin levels. These molecules were associated with neonatal gut integrity and maturation, notably by acting on tight junctions. Their significant changes in miR-30b milk could be clearly involved in the early intestinal closure phenotype of the pups, in connection with the observed early growth defect. Further investigations are now needed to determine their specific mode of action, with the aim to modulate infant diet in regard with a benefic effect on growth and health.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Milk

SUBMITTER: Céline Henry

LAB HEAD: Sandrine Le Guillou

PROVIDER: PXD037629 | Pride | 2023-10-09

REPOSITORIES: Pride

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			Action	DRS
	20170203_02_Strata26WT.raw	Raw
	20170203_02_Strata26WT.xml	Xml
	20170203_04_Strata29TG.raw	Raw
	20170203_04_Strata29TG.xml	Xml
	20170203_06_Strata18WT.raw	Raw

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Project description:Human milk (breastmilk) is much more than nutrition for the infant, containing an array of regulatory agents with immunoprotective and developmental functions. Amongst those, microRNAs (miRNAs) have recently been identified, with their properties, roles, origin and distribution in breastmilk as well as in the mammary gland being still undetermined. In this study, we examined the miRNA profile of different fractions of human milk (cells and lipids) using the OpenArray system (Applied Biosystems, 770 miRNA species measured per sample) and compared it with maternal peripheral blood mononuclear cells (PBMCs) and plasma. Although PBMCs were the richest group in miRNA species, plasma showed very low expression pattern. Thus, the human milk fractions (cells, lipid) and skim milk (not being investigated in this study) were found to conserve higher levels of miRNAs than blood in general. Specifically, human milk cell miRNA quantity was found relatively close to PBMCs, and higher than milk lipids. Correlation and clustering analyses indicated that miRNA expression and types of milk cells were highly similar to those in lipids. Milk miRNAs showed a slight correlation to PBMCs, so PBMCs potentially are not contributing to milk miRNAs. Plasma was different to all other three groups in miRNA content and expression pattern. Further, two infant formulae (a plant-based and a cow milk-based) were compared to human milk and found to contain significantly fewer miRNA species than human milk cells and lipids (p>0.001). Taken together with previous studies on miRNAs, our findings demonstrate that human milk is one of the richest sources of miRNAs among human body fluids. As a non-invasive and plentiful source of miRNAs, human milk could be used as a disease biomarker for the mammary gland, with potential in assessing lactation performance. Finally, gene target and pathways analyses identified several target mRNAs regulated by miRNAs found to be abundant in breastmilk. Given the recently identified stability and function of food-derived miRNAs in regulating mammalian genes, we propose that breastmilk is a rich source of miRNA ingested by the infant during the first months of life, and which potentially contribute to early infant development. 10 exclusively breastfeeding dyads were recruited. 10 whole milk and 10 whole blood samples were collected and fractionated to obtain 10 milk cells, 10 milk lipid, 10 mononeucleoted blood cells (PBMCs), and 10 plasma. In addition to the above 40 samples, 2 infant formula were profiled. 4 different extraction kits were used, miRNeasy mini Kit for human milk cell and PBMC samples. miRCURY RNA Isolation-Biofluids Kit for human milk lipid samples and both infant formulae. mirVana PARIS Kit for plasma samples. NanoDrop 2000 and Bioanalyzer 2100 were used to determine concentration and purity of the extracted miRNA from all samples (n=42). miRNA OpenArray panel system (Life Technologies, CA, USA) was used to profile 754 human mature miRNAs in samples. RNU48, RNU44 and U6 rRNA were used as housekeeping controls for normalisation. ath-miR159a was used as a negative control for human samples. GeneGO and Ingenuity Pathway Analysis were used to determine biological pathways. Please note that normalization of miRNAs was done in R but without generating deltaCT values, thus [1] only the list of normalized miRNA with Ct vlaue between 8 and 29 and that detected in at least 4 samples out of 10 analysed in each group is provided ('normalized_miRNAs_list.txt') [2] the sample data tables contain raw data.

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Specific milk composition in miR-30b-transgenic mice associated to offspring early gut maturation with lasting consequences on their growth

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