Unknown,Transcriptomics,Genomics,Proteomics

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ScRNA-seq analysis of human mammary epithelial cells derived from non-lactating breast tissue or from milk

ABSTRACT: Breast cancer risk is influenced by parity in an age-dependant manner, however human tissue remodelling induced by pregnancy and lactation is not well understood. In most cases, it is difficult to acquire human breast tissue during these key stages of development. Here, we present an approach to overcome this using single-cell RNA sequencing to examine viable primary mammary epithelial cells isolated from human milk (n=4) compared to resting, non-lactating human breast tissue (n=4). Overall, we examined 54,323 viable high-quality cells from mixed age and parous individuals consisting of 29,078 lactation derived mammary cells (LMCs) and 25,245 non-lactation derived mammary cells (NMCs). We identified all documented mammary subpopulations within our breast tissue samples and found that milk contains distinct secretory luminal cells together with myeloid and lymphocytic hematopoietic lineage cells. Comparing the luminal transcriptional profiles of cells isolated from the resting and lactating state identified differences in mammary cell function and metabolism between these maturation states. In future this data may be used to provide an insight into how parity influences human luminal cell metabolism and breast cancer risk.

INSTRUMENT(S): 10x Genomics Chromium Controller, Illumina NovaSeq 6000

ORGANISM(S): Homo sapiens

SUBMITTER: Alecia-Jane Twigger

PROVIDER: E-MTAB-9841 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Background: The mammary gland is a dynamic organ that undergoes important physiological changes during reproductive cycles. Until now, data regarding the characterisation of miRNA in the mammary gland have been scarce and mainly focused on their abnormal expression in breast cancer. Our goal was to characterise the microRNA (miRNA) involved in mechanisms regulating the mammary function, with particular focus on the lactation stage Methodology/principal findings: Using high-throughput sequencing technology, an exhaustive repertoire of miRNA (miRNome) expressed in mouse and bovine mammary glands during established lactation was identified, characterised and compared. Furthermore, in order to obtain more information on miRNA loading in the RNA-induced silencing complex (RISC), the miRNome was compared with that obtained from RNA associated with the AGO2 protein (AGO2-miRNome) in mouse lactating mammary gland. This study enabled the identification of 164 and 167 miRNA in mouse and bovine, respectively. Among the 30 miRNA most highly expressed in each species, 24 were common to both species and six of them tended to be more highly expressed in lactating than in non-lactating mammary gland. The potential functional roles of these 24 miRNA were deduced using DIANA-miRPath software, based on miRNA/mRNA interactions. Moreover, seven putative novel miRNA were identified. Using DAVID analysis, it was concluded that the predicted targets of two of these putative novel miRNA are involved in mammary gland morphogenesis. Conclusion/significance: Our study provides an overview of the characteristics of lactating mouse and bovine mammary gland miRNA expression profiles. Moreover, species-conserved miRNA involved in this fundamental biological function were identified. These miRNomes will new be used as references for further studies during which the impact of animal breeding on the miRNA expression will be analysed. microRNA profiles of mammary glands from 2 FVB/N mice at lactation day-12 and mammary biopsies from 4 Holstein dairy cows at mid-lactation, generated by deep sequencing, using Illumina HiSeq 2000.

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ScRNA-seq analysis of human mammary epithelial cells derived from non-lactating breast tissue or from milk

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