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Drop-seq analysis of bleomycin-treated and control mouse lung single-cell suspensions

ABSTRACT: Drop-seq analysis of bleomycin-treated and control mouse lung single-cell suspensions

PROVIDER: PRJNA437767 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6823950_1.fastq.gz	Fastqsanger.gz
	SRR6823950_2.fastq.gz	Fastqsanger.gz
	SRR6823951_1.fastq.gz	Fastqsanger.gz
	SRR6823951_2.fastq.gz	Fastqsanger.gz
	SRR6823952_1.fastq.gz	Fastqsanger.gz

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Drop-seq analysis of bleomycin-treated and control mouse lung single-cell suspensions

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Drop-seq analysis of bleomycin-treated and control mouse lung single-cell suspensions

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Drop-seq analysis of bleomycin-treated and control mouse lung single-cell suspensions

Project description:Drop-seq analysis of bleomycin-treated and control mouse lung single-cell suspensions

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Longitudinal single-cell transcriptomics analysis of mouse lung upon bleomycin-induced injury

Project description:Two single-cell RNA sequencing data sets were generated called "Whole lung" and "High Resolution". The "Whole lung" single-cell mRNAseq libraries were generated with Drop-Seq from whole mouse lungs upon bleomycin-induced injury and followed over time. Samples were taken at days 3 (n = 3), 7 (n = 5), 10 (n = 3), 14 (n = 4), 21 (n = 4) and 28 (n = 2). Control samples (n = 7) were administered saline only, also indicated with PBS or day0. The "High resolution" single-cell mRNAseq libraries were generated with Drop-Seq from the epithelial compartment of mouse lungs upon bleomycin-induced injury and followed over time. Samples were taken daily for two weeks and at days 21, 28, 36, 54 after injury. Control samples (n = 2) were administered saline only, also indicated with PBS or day0.

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Project description:The molecular mechanisms of lung injury and fibrosis are incompletely understood. microRNAs (miRNAs) are crucial biological regulators by suppression of their target genes and are involved in a variety of pathophysiologic processes. To gain insight into miRNAs in the regulation of lung fibrosis, total RNA was isolated from lung samples harvested at different days after bleomycin treatment, and miRNA array was performed thereafter. miRNAs expressed in lungs with bleomycin treatment at different time points were compared to miRNAs expressed in lungs without bleomycin treatment, resulting in 161 miRNAs differentially expressed. Furthermore, miRNA expression patterns regulated in initial and late periods after bleomycin were identified. Target genes were predicted in silico for differentially expressed miRNAs, including miR-7f, miR-7g, miR-196b, miR-16, miR-195, miR-25, miR-144, miR-351, miR-34a, miR-499, miR-704, miR-717, miR-10a, miR-211, miR-34a, miR-367 and miR-21, and then cross-referenced to molecular pathways including apoptosis, Wnt, Toll-like receptor, and TGF-? signaling, which are involved in different pathological phenotypes such as apoptosis, inflammation, and fibrosis. Our study demonstrated relative abundance of miRNA levels in bleomycin-induced lung fibrosis. The miRNAs and their potential target genes identified herein contribute to the understanding of the complex transcriptional program of lung fibrosis. Under anesthesia, 2.5 U/kg bleomycin dissolved in sterile PBS was administered via trachea as previously described. Lung tissues were harvested at the time point of day 0, 3, 7, 14, and 21 post bleomycin challenges. 3 sample in specific time point, except for day 14 where nday14 = 2.

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Project description:We perfomed a microRNA microarray on to assess differentially expressed miRNAs in bleomycin-induced lung fibrosis in mice. To induce pulmonary fibrosis, belomycine (Sigma-Aldrich, St Louis, MO) was dissolved and administred intratracheally at a dose of 1.5U/kg body weight. Control animals received saline only. 21 days post bleomycin treatment, mice were sacrificed and lung tissue were collected for RNA extraction and microRNA microarray.

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