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Targeted immunotherapy rescues pulmonary fibrosis by reducing activated fibroblasts and regulating alveolar cell profile

ABSTRACT: The single-cell RNA-seq libraries were generated with 10x genomics from mouse lungs. Model mice were treated with saline (n=2) or transient CAR-T cells generated by LNP in vivo (n=2) after bleomycin-induced injury. The samples were taken at day28 after surgery and collected as the fresh whole lung tissue.

ORGANISM(S): Mus musculus

PROVIDER: GSE283267 | GEO | 2025/03/19

REPOSITORIES: GEO

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Targeted immunotherapy rescues pulmonary fibrosis by reducing activated fibroblasts and regulating alveolar cell profile

Project description:Idiopathic pulmonary fibrosis (IPF) is a severe lung disease in the world, but has limited clinical therapies. Fibrosis maintains dynamic balance with overactivated fibroblasts. Given pulmonary cell regeneration, the lung may have self-repairing ability if fibrosis is removed by clearance of overactivated fibroblasts. The aim of the study is to evaluate therapeutic activity of transient CAR-T cells generated via a novelly designed LNP-mRNA system and address the relevant mechanism to epithelial cell polarization in a mouse model of bleomycin-induced IPF. Studies on proteomes and histology showed that fibrosis-induced ECM stiffening impaired epithelial cell polarization that limited cell’s self-healing ability. The proposed LNP-mRNA therapy eliminated overactivated fibroblasts and removed pulmonary fibrosis successfully. The decreased ECM stiffness and the improvement of extracellular environment facilitated re-establishment of cell polarity and restored the self-repairing ability of epithelial cell. Hence, LNP-mRNA treatment for IPF can recover pulmonary structure and function close to a healthy lung. This therapy shows a potential treatment for IPF patients.

2025-03-19 | PXD053333 | Pride

RNA-Seq analysis of alveolar macrophages from normal and fibrotic mouse lungs

Project description:We performed gene expression profiling of alveolar macrophages from mice that were intratracheally instilled with saline or bleomycin. We identified a number of differentially expressed genes in the two groups of cells.

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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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Bleomycin induces molecular changes directly relevant to idiopathic pulmonary fibrosis: A model for “active” disease.

Project description:Genomic profiling of bleomycin- and saline-treated mice across 7 timepoints (1, 2, 7, 14, 21, 28, 35 days post treatment) was carried out in C57BL6/J mice to determine the phases of response to bleomycin treatment which correspond to onset of active pulmonary fibrosis. Temporal genomic characterization of lung homogenate from male C57BL6/J mice treated intratracheally with bleomycin or saline was carried out at 7 timepoints post treatment (1, 2, 7, 14, 21, 28, 35 days). Bleomycin (2U/kg) in 50 μl was intratracheally sprayed once into mice lightly anaesthetized with isoflurane (5% in 100% O2). Control animals received 50 μl of saline. Total RNA was isolated from the mouse lung tissue of bleomycin- and saline-treated mice across the 7 time points (n=8 per group) and homogenized in QIAzol reagent. Purified total RNA was amplified and labeled using NuGen Ovation kits (NuGEN Technologies, Inc., San Carlos, CA), and RNA from samples was hybridized to Affymetrix Mouse 430 2.0 arrays. One sample (saline treated, d14) was flagged as an outlier in principal component analysis and removed from subsequent analysis.

2013-04-10 | E-GEOD-40151 | biostudies-arrayexpress

Targeted immunotherapy rescues pulmonary fibrosis by reducing activated fibroblasts and regulating alveolar cell profile

Project description:Targeted immunotherapy rescues pulmonary fibrosis by reducing activated fibroblasts and regulating alveolar cell profile

| PRJNA1192992 | ENA

Higher susceptibility of males to bleomycin-induced pulmonary inflammation is associated with sex-specific transcriptomic differences in myeloid cells

Project description:In this study, we subjected male and female mice to bleomycin-induced lung injury for 14-days. Bronchoalveolar lavage cells were isolated and subjected to RNA-sequencing. Four groups were compared. Saline-Males and females and bleomycin-males and females.

2022-06-10 | GSE198631 | GEO

Murine lung transcripts during soluble egg antigen (SEA)-induced allergic asthma and bleomycin-induced pulmonary fibrosis in Wildtype, IL-13Ra1 and IL-13Ra2 deficient mice sensitized and challenged

Project description:Transcriptional profiling of mouse lungs by comparing PBS and saline treated lungs with SEA and bleomycin treated lungs, respectively. Mice strains used in this study include wild type (C57BL/6), IL-13Ra1 and IL-13Ra2 deficient mice. In mouse model of allergic asthma, mice were sensitized twice (day0 and day14) by i.p. injection of 10 µg of SEA. On days 28 and 31 mice were anesthetized with a mixture of xylazine and ketamine and given an intratracheal (I.T) airway challenge with 10 µg of SEA. Mice were sacrificed 24 h after the final airway challenge (day 32) and lungs were collected for RNA preparation. Biological replicates include 5 PBS treated and 5 SEA treated mice. In bleomycin-induced fibrosis odel, mice were anaesthetized with a xylazine and ketamine cocktail and given 0.15 U bleomycin sulfate (EMD) in saline or saline alone via the I.T. route. On day 7, mice were sacrificed and ungs were collected for RNA preparation. Biological replicates include 4 saline treated and 4 bleomycin treated mice. Goal was to determine IL-13 receptors regulated genes during SEA induced allergic asthma and bleomycin-induced fibrosis. Fluorescent cDNA targets were prepared from a 20 μg experimental RNA sample (SEA or bleomycin challenged group–dUTPCy5 – Amersham, Piscataway, NJ) and a 20 μg reference RNA sample (PBS or saline treated group–dUTPCy3- Amersham, Piscataway, NJ). Equal quantities of the above labeled cDNA (experimental and reference labeled RNA samples) were mixed and any free label present in the sample was removed by washing 3 times using a 10 kDa cutoff Vivaspin filters (Millipore). Labeled fluorescent cDNA targets were hybridized on the Whole Mouse Genome Oligo Microarray Kit (Agilent, Palo Alto, CA) containing more than 41000 gene probes.

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GLI1+ mesenchymal stromal cells modulate epithelial metaplasia in lung fibrosis

Project description:A single-cell transcriptional analysis was performed on GLI1+ stromal cells from the adult murine lung during homeostasis and after fibrotic injury. The goal is to understand the role of GLI1+ stromal cells in lung fibrosis and repair. Whole adult murine lung tissue from two samples were separately dissociated to single cells and subjected to fluorescence activated cell sorting (FACS) to select all live GLI1+ cells. One sample was treated with bleomycin to induce fibrosis and the other was treated with saline as a control. The single cell RNA-sequencing library was generated separately for the bleomycin and saline-treated samples. Cells were sequenced at a depth of ~70,000 reads/cell. We captured approximately 17,700 cells with a median of 2,400 genes detected per cell utilizing a droplet-based barcoding approach to capture single cells for RNA sequencing. After bleomycin-induced fibrosis, we identified a novel "myofibroblast" subset of GLI1 cells that contribute to injury and repair. Injured GLI1 cells also reveal dysregulation in key developmental pathways, including BMP signaling, that contribute to metaplastic repair after fibrotic injury.

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Bleomycin induces molecular changes directly relevant to idiopathic pulmonary fibrosis: A model for “active” disease.

2013-04-10 | GSE40151 | GEO

Human platelet lysate enhances in vivo activity of CAR-Vδ2 T cells by reducing cellular senescence and apoptosis

Project description:Vγ9Vδ2 T cells are an attractive cell platform for the off-the-shelf cancer immunotherapy due to their lack of alloreactivity and inherent multi-pronged cytotoxicity, which could be further amplified with chimeric antigen receptors (CARs). In this study, we sought to enhance the in vivo longevity of CAR-Vδ2 T cells by modulating ex vivo manufacturing conditions and selecting an optimal CAR costimulatory domain. Specifically, we compared the anti-tumor activity of Vδ2 T cells expressing anti-CD19 CARs with costimulatory endodomains derived from CD28, 4-1BB or CD27 and generated in either standard fetal bovine serum (FBS)- or human platelet lysate (HPL)-supplemented medium. We found that HPL supported greater expansion of CAR-Vδ2 T cells with comparable in vitro cytotoxicity and cytokine secretion to FBS-expanded CAR-Vδ2 T cells. HPL-expanded CAR-Vδ2 T cells showed enhanced in vivo anti-tumor activity with longer T cell persistence compared to FBS counterparts, with 4-1BB costimulated CAR showing the greatest activity. Mechanistically, HPL-expanded CAR Vδ2 T cells exhibited reduced apoptosis and senescence transcriptional pathways compared to FBS-expanded CAR-Vδ2 T cells and increased telomerase activity. This study supports enhancement of therapeutic potency of CAR-Vδ2 T cells through a manufacturing improvement.

2024-08-16 | GSE247760 | GEO

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