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ChIP-seq Analysis for H3K4me3 in Neomycin-induced Lgr5+ Progenitor Cells of Neonatal Mouse Cochlea

ABSTRACT: ChIP-seq Analysis for H3K4me3 in Neomycin-induced Lgr5+ Progenitor Cells of Neonatal Mouse Cochlea

PROVIDER: PRJNA735199 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14730718.fastq.gz	Fastqsanger.gz
	SRR14730719.fastq.gz	Fastqsanger.gz
	SRR14730720.fastq.gz	Fastqsanger.gz
	SRR14730721.fastq.gz	Fastqsanger.gz
	SRR14730722.fastq.gz	Fastqsanger.gz

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ChIP-seq Analysis for H3K4me3 in Neomycin-induced Lgr5+ Progenitor Cells of Neonatal Mouse Cochlea

Project description:In the study, we detailedly manifested the regulatory roles of histone modifications in neomycin-induced proliferation of Lgr5+ progenitor and HC regeneration by integrated RNA-seq and ChIP-seq analysis. we found 94 differentially expressed genes which are related to differential binding regions via analyzing ChIP-seq and RNA-seq data. we are delighted to find that H3K4me3 can modulate the expressions of 12 genes to involve in cell cycle, neural development, inner ear development, Wnt and Hippo signaling pathways to regulate proliferation and HC regeneration based on the protein interaction network and gene ontology (GO) functional annotation analysis.

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Characterization of Lgr5+ Progenitor Cell Transcriptomes after Neomycin Injury in the Neonatal Mouse Cochlea.

Project description:Lgr5+ supporting cells (SCs) are enriched hair cell (HC) progenitors in the cochlea. Both in vitro and in vivo studies have shown that HC injury can spontaneously activate Lgr5+ progenitors to regenerate HCs in the neonatal mouse cochlea. Promoting HC regeneration requires the understanding of the mechanism of HC regeneration, and this requires knowledge of the key genes involved in HC injury-induced self-repair responses that promote the proliferation and differentiation of Lgr5+ progenitors. Here, as expected, we found that neomycin-treated Lgr5+ progenitors (NLPs) had significantly greater HC regeneration ability, and greater but not significant proliferation ability compared to untreated Lgr5+ progenitors (ULPs) in response to neomycin exposure. Next, we used RNA-seq analysis to determine the differences in the gene-expression profiles between the transcriptomes of NLPs and ULPs from the neonatal mouse cochlea. We first analyzed the genes that were enriched and differentially expressed in NLPs and ULPs and then analyzed the cell cycle genes, the transcription factors, and the signaling pathway genes that might regulate the proliferation and differentiation of Lgr5+ progenitors. We found 9 cell cycle genes, 88 transcription factors, 8 microRNAs, and 16 cell-signaling pathway genes that were significantly upregulated or downregulated after neomycin injury in NLPs. Lastly, we constructed a protein-protein interaction network to show the interaction and connections of genes that are differentially expressed in NLPs and ULPs. This study has identified the genes that might regulate the proliferation and HC regeneration of Lgr5+ progenitors after neomycin injury, and investigations into the roles and mechanisms of these genes in the cochlea should be performed in the future to identify potential therapeutic targets for HC regeneration.

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Cochlea Neomycin Timecourse

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Project description:Our previous studies showed that Lgr6+ cells are a subpopulation of Lgr5+ progenitor cells and that both Lgr6+ and Lgr5+ progenitors can regenerate Myosin7a+ HCs in vitro. Thus we hypothesized that Lgr6+ cells are an enriched population of cochlear progenitor cells. However, the detailed distinctions between the Lgr5+ and Lgr6+ progenitors have not been researched. Here, we systematically compared the proliferation, HC differentiation, and detailed transcriptome expression profiles of these two progenitor populations. We found that when isolated by flow cytometry, the same number of Lgr6+ progenitors generated significantly more Myosin7a+ HCs than Lgr5+ progenitors; however, Lgr5+ progenitors formed more epithelial colonies and more spheres than Lgr6+ progenitors in vitro. Using RNA-Seq, we compared the transcriptome differences between Lgr5+ and Lgr6+ progenitors and identified a list of significantly differential expressed genes which may regulate the proliferation and differentiation of these HC progenitors.

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