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RNA-sequencing comparison of pneumonectomy and bleomycin reveals novel anti-fibrotic mechanisms and regulators

ABSTRACT: RNA-sequencing comparison of pneumonectomy and bleomycin reveals novel anti-fibrotic mechanisms and regulators

PROVIDER: PRJNA677893 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13040130_1.fastq.gz	Fastqsanger.gz
	SRR13040130_2.fastq.gz	Fastqsanger.gz
	SRR13040131_1.fastq.gz	Fastqsanger.gz
	SRR13040131_2.fastq.gz	Fastqsanger.gz
	SRR13040132_1.fastq.gz	Fastqsanger.gz

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RNA-sequencing comparison of pneumonectomy and bleomycin reveals novel anti-fibrotic mechanisms and regulators

Project description:Fibroblast activation is transient in successful wound repair, but persistent in fibrotic pathologies. Understanding fibroblast deactivation during successful wound healing may provide new approaches to therapeutically reverse fibrogenic cell activation. To characterize the gene programs that accompany fibroblast activation and reversal during lung fibrosis resolution, we used RNA-seq analysis of flow-sorted Col1α1-GFP positive and CD45, CD31, and CD326 negative cells isolated from the lungs of young mice exposed to bleomycin and pneumonectomy. We compared fibroblasts isolated from controls to those isolated at days 14 and 30 after bleomycin exposure, as well as at 14 days after pneumonectomy, representing the peak of extracellular matrix deposition, early stage of fibrosis resolution, and non-fibrotic lung repair respectively.

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Project description:Wildtype mice were given saline or bleomycin by oropharyngeal instillation. After 14 days, during the fibrotic phase of the response, lungs were dissected and total RNA was extracted and used for gene expression profiling. The aim was to identify those genes regulated during the development of fibrosis in this animal model of bleomycin-induced lung fibrosis. Wildtype male C57Bl/6J mice (8-10 weeks old) were used in the study, with three mice per group. Bleomycin (1mg/kg body weight in 50μl of saline) or saline was administered by oropharyngeal installation as described previously by Lakatos et al, Exp Cell Res, 2006, under light halothane-induced anaesthesia. After 14 days, lungs were removed, blotted dry and the trachea and major airways were excised before the separated lobes were snap frozen in liquid nitrogen. Lungs were then pulverized under liquid nitrogen, and the resulting lung powder was used for total RNA extraction using Trizol. Following DNase-treatment, clean-up, cDNA synthesis and cRNA synthesis, samples were hybridized to Affymetrix MOE430A genechips using standard protocols. The data were analyzed using RMA with quantiles normalization.

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Transcriptional profile of mouse lung during the fibrotic phase of the response to bleomycin

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Comparison of gene change after pneumonectomy

Project description:Comparison of gene change after pneumonectomy

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RNA-seq profiling of ITPKC overexpression in normal human lung fibroblasts

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