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Effect of neutrophil depletion on gene expression im macrophages and fibroblasts from mouse lung and skin

ABSTRACT: Effect of neutrophil depletion on gene expression im macrophages and fibroblasts from mouse lung and skin

PROVIDER: PRJNA837203 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR19167556_1.fastq.gz	Fastqsanger.gz
	SRR19167556_2.fastq.gz	Fastqsanger.gz
	SRR19167557_1.fastq.gz	Fastqsanger.gz
	SRR19167557_2.fastq.gz	Fastqsanger.gz
	SRR19167558_1.fastq.gz	Fastqsanger.gz

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Effect of neutrophil depletion on gene expression im macrophages and fibroblasts from mouse lung and skin

Project description:To investigate the effect of the absence of neutrophils in the transcriptome of fibroblast and macrophage from lung and skin we injected DT to Mrp8 CRE+ iDTR and Mrp8 CRE- iDTR mice every two days for a week and isolated lung and skin fibroblasts and macrophages by cell sorting . We then performed gene expression profiling analysis using data obtained from RNA-seq of macrophages and fibroblast 7 days after DT treatment and compared the transcriptomes of macrophages and fibroblasts in control or neutrophil depletion animals. We focused on the role of neutrophils on the regulation of extracelular matrix genes in fibroblasts and macrophages and found that neutrophil depletion does not significantly affect the RNA level of matrix-related transcripts in those cells.

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Project description:To elucidate the role of neutrophils in the molecular alterations observed after I/R, we performed an experiment of I/R in neutrophil-depleted mice. Neutrophil depletion was performed by anti-Ly6G injection and IgG was used as vehicle. We have performed an unbiased, open-search analysis of the oxidative PTMs and quantitative multiplexed proteomics that take place in the mouse heart tissue at 24h after I/R. Neutrophil depletion was performed in 3 mice by Ly6G injection 24h and 48 h prior to I/R and sacrificed 24 hours after reperfusion. A second group consisting on 3 mice that were injected with IgG antibody as vehicle, and subjected to I/R and sacrificed 24h after reperfusion. A third group of 3 mice with no injection were also sacrificed 24h after reperfusion. 12 additional mice from the three previous groups were sacrificed without I/R intervention: control without injection (n=6), IgG injection (n=3) and Ly6G injection (n=3). Whole hearts tissue was collected for proteomics evaluation.

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Single-cell RNAseq analysis (10X Genomics Chromium) of refilled lung interstium macrophages on day 4 after depletion

Project description:Lung interstitium macrophages (IMs) are non-alveolar resident tissue macrophages which contribute to the lung homeostasis. These cells were reported to be heterogeneous by our group and other teams, which contains two main distinct subpopulations: CD206+ IMs and CD206- IMs. However, the exact origin of IMs and the transcriptional programs that regulate IM differentiation remains unclear. In recent report, we analyzed the refilled IMs in the course of time after induced IM depletion with single-cell RNA sequencing (10X Genomics Chromium) and bulk RNA sequencing. The IMs in Day 4 post-depletion were compared to the those without depletion. Results showed that refilled IMs had a lower ratio of CD206+ IM vs CD206- subpopulation comparing to IMs without depletion, but they shared high similarity to each other, indicating that the de novo IM population had been established before Day 4 post-depletion.

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Project description:Neutrophils patrol healthy tissues and confer local defense upon perturbations. Recent evidence suggests that they participate in tissue-specific structural programs in addition to their indispensable role in immunity.The goal of this study is to evaluate the effect of neutrophils in healthy mouse skin by comparing the transcriptional landscape of naive skin to neutrophil-depleted skin.

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Project description:Neutrophils patrol healthy skin at steady state. Here we evaluate whether the antibody-mediated depletion of neutrophils affect global transcriptome of the skin.

2025-01-09 | GSE229493 | GEO

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