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A2AR-eGFP reporter mouse enables elucidation of A2AR expression dynamics during anti-tumor immune responses

ABSTRACT: A2AR-eGFP reporter mouse enables elucidation of A2AR expression dynamics during anti-tumor immune responses

PROVIDER: PRJNA957714 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24225667.fastq.gz	Fastqsanger.gz
	SRR24225668.fastq.gz	Fastqsanger.gz
	SRR24225669.fastq.gz	Fastqsanger.gz
	SRR24225670.fastq.gz	Fastqsanger.gz
	SRR24225671.fastq.gz	Fastqsanger.gz

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Similar Datasets

A2AR-eGFP reporter mouse enables elucidation of A2AR expression dynamics during anti-tumor immune responses

Project description:Understanding of the mechanism by which A2AR is regulated has been limited by difficulty in identifying the cell types that express A2AR due to a lack of robust antibodies for these receptors. To overcome this we developed an A2AR-eGFP reporter mouse and assessed the expression of A2AR during ongoing anti-tumor immune responses.

2023-10-03 | GSE230135 | GEO

Role of Adenosine 2A Receptors (A2AR) on regulatory T cells (Tregs)

Project description:The adenosine 2A receptor (A2AR) is expressed on regulatory T cells (Tregs), but the functional significance is currently unknown. We compared the gene expression between wild-type (WT) and A2AR knockout (KO) Tregs and between WT Tregs treated with vehicle or a selective A2AR agonist. FACS-sorted GFP positive Tregs from WT or A2AR KO FoxP3GFP mouse spleen and lymph nodes were incubated 18 hr with vehicle (DMSO), a separate set of WT Tregs were incubated with the selective A2AR agonist ATL1222 10 nM (Dogwood Pharmaceuticals, Inc.) for 18 hr prior to RNA isolation.

2012-05-31 | E-GEOD-34006 | biostudies-arrayexpress

scRNA-seq of lung CD140a+ mesenchymal cells (fibroblasts) from CD140a-EGFP reporter mice

Project description:To investigate the heterogeneity of lung CD140a+ mesenchymal cells (fibroblasts) and identify the specific lung fibroblast subset, we purified CD140a+ lung cells using CD140a-EGFP reporter mice for a higher-resolution scRNA-seq.

2022-07-19 | E-MTAB-11875 | biostudies-arrayexpress

Gene expression in haematopoietic stem and progenitor cells from Gata1-EGFP reporter mouse bone marrow

Project description:Haematopoietic stem cells can differentiate into all blood cell types. In this process, cells become progressively restricted to a single cell type. The order in which differentiating cells loose lineage potential, and the prospective isolation of cells with a defined potential remains a long-standing question. We performed gene expression analysis of haematopoietic cells from Gata1-EGFP reporter mice, leading to a model for hematopoiesis where the initial lineage decision consists of a separation of erythroid/megakaryocyte/mast cell/eosinophil potential from lymphopoietic/monocyte/neutrophil potential RNA was isolated from HSC EGFP-, HSC EGFP+, LMPP, CLP, preGM EGFP-, preGM EGFP+, GMP EGFP-, GMP EGFP+ and preMegE cells with the QIAGEN RNeasy Micro Kit.

2016-04-04 | E-GEOD-49241 | biostudies-arrayexpress

Role of Adenosine 2A Receptors (A2AR) on regulatory T cells (Tregs)

2012-06-01 | GSE34006 | GEO

Effect of blocking A2aR in sepsis on the Treg transcriptome

Project description:In our study, blocking A2aR with ZM241385 was found to inhibit Foxp3 expression in Treg cells of septic mice. Therefore, we used flow cytometry to sort mouse spleen CD4+CD25+ cells for transcriptome sequencing in an attempt to discover the mechanism by which blocking A2aR affects Foxp3 expression. The specific groupings were as follows: naive groups (N1-N5), CLP groups (C1-C5), CLP+A2aR antagonist groups (Z1-Z6).

2023-10-03 | GSE244227 | GEO

A<sub>2A</sub>R eGFP reporter mouse enables elucidation of A<sub>2A</sub>R expression dynamics during anti-tumor immune responses.

Project description:There is significant clinical interest in targeting adenosine-mediated immunosuppression, with several small molecule inhibitors having been developed for targeting the A2AR receptor. Understanding of the mechanism by which A2AR is regulated has been hindered by difficulty in identifying the cell types that express A2AR due to a lack of robust antibodies for these receptors. To overcome this limitation, here an A2AR eGFP reporter mouse is developed, enabling the expression of A2AR during ongoing anti-tumor immune responses to be assessed. This reveals that A2AR is highly expressed on all tumor-infiltrating lymphocyte subsets including Natural Killer (NK) cells, NKT cells, γδ T cells, conventional CD4+ and CD8+ T lymphocytes and on a MHCIIhiCD86hi subset of type 2 conventional dendritic cells. In response to PD-L1 blockade, the emergence of PD-1+A2AR- cells correlates with successful therapeutic responses, whilst IL-18 is identified as a cytokine that potently upregulates A2AR and synergizes with A2AR deficiency to improve anti-tumor immunity. These studies provide insight into the biology of A2AR in the context of anti-tumor immunity and reveals potential combination immunotherapy approaches.

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scRNA-seq of lung CD140a+ mesenchymal cells (fibroblasts) from CD140a-EGFP reporter mice

Project description:scRNA-seq of lung CD140a+ mesenchymal cells (fibroblasts) from CD140a-EGFP reporter mice

| PRJEB54002 | ENA

Generation of a TPH2-EGFP reporter cell line for Purification and Monitoring of Human Serotonin neurons in vitro and in vivo

Project description:Generation of serotonin neurons (SNs) from human pluripotent stem cells (hPSCs) provides a promising platform to explore the mechanisms of serotonin-associated neuropsychiatric disorders. However, neural differentiation always yields heterogeneous cell populations, making it difficult to identify and purify SNs in vitro or track them in vivo following transplantation. Herein, we generated a TPH2-EGFP reporter hPSC line with insertion of EGFP into the endogenous tryptophan hydroxylase 2 (TPH2) locus using CRISPR-Cas9-mediated gene editing technology. This TPH2-reporter, which faithfully indicated TPH2 expression during differentiation, enabled us to obtain purified SNs for subsequent transcriptional analysis and study of pharmacological responses to antidepressants. In addition, the reporter system showed strong EGFP expression to indicate SNs, which enabled us to explore in vitro and ex vivo electrophysiological properties of SNs. In conclusion, this TPH2-EGFP reporter cell line might be of great significance for studies on human SN-related development and differentiation, drug screening, disease modeling, and cell replacement therapies.

2022-09-22 | GSE206341 | GEO

Bulk RNA-Seq analysis of TCF1 Reporter eGFP+ and TCF1 Reporter eGFP- CD8+ T cells generated with different metabolic manipulations and isolated from in vitro co-culture with HKP1-ova-GFP mouse lung adenocarcinoma cells

Project description:To investigate how metabolic manipulation alters differentiation and the transcriptional landscape in tumor-specific CD8+ T cells, we isolated OT-I+ CD8+ T cells from spleens of transgenic OT-I+ Tcf7GFP+ mice expressing a TCF1 eGFP Reporter, and activated them for one day on plates coated with 2μg/ml of both anti-CD3 and anti-CD28 in the presence of 50U/mL mouse IL-2. We then treated them with equal volumes of DMSO vehicle control, 3μM glucose 6-phosphate dehydrogenase agonist AG1, 25μM glucose 6-phosphate dehydrogenase inhibitor G6PDi-1, or 2mM hexokinase inhibitor 2-DG, and expanded them for 1 more day on anti-CD3/anti-CD28 coated plates in the presence of 50U/mL mouse IL-2. We subsequently co-cultured these cells with HKP1-ova-GFP tumor cells at a 5:1 T cell:tumor cell ratio for an additional 4 days with continuing drug treatment in the presence of 50U/mL mouse IL-2, passing cultures every other day. At day 6 post-initial-stimulation, CD8+ T cells were harvested, filtered by passing through a 70uM cell strainer, washed, and stained for CD8β and Thy1.2. Samples were dyed with DAPI at 0.2μg/mL, and sorted on a Becton-Dickinson FACS Aria II sorter for DAPI- CD8β + Thy1.2+ eGFP+ or DAPI- CD8β + Thy1.2+ eGFP- cells. We then performed gene expression profiling analysis using data obtained from RNA-seq of sorted cells, comparing across treatment groups and TCF1 eGFP Reporter expression.

2023-10-19 | GSE238203 | GEO

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