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Bulk RNAseq analysis of CD4 CD8 double negative 3 (DN3) and CD4 CD8 double positive (DP) thymocytes in thymus autonomy and turnover

ABSTRACT: Bulk RNAseq analysis of CD4 CD8 double negative 3 (DN3) and CD4 CD8 double positive (DP) thymocytes in thymus autonomy and turnover

PROVIDER: PRJNA668067 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12793669.fastq.gz	Fastqsanger.gz
	SRR12793670.fastq.gz	Fastqsanger.gz
	SRR12793671.fastq.gz	Fastqsanger.gz
	SRR12793672.fastq.gz	Fastqsanger.gz
	SRR12793673.fastq.gz	Fastqsanger.gz

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Bulk RNAseq analysis of CD4 CD8 double negative 3 (DN3) and CD4 CD8 double positive (DP) thymocytes in thymus autonomy and turnover

Project description:Wild type thymi were transplanted into wild recipients (turnover) or Rag2-/-gc-/- mice (autonomy) and, after 28 days of transplant, DN3 and DP thymocytes were sorted and their transcriptomes analyzed.

2021-04-12 | GSE159218 | GEO

Single cell RNAseq coupled with TCR repertoire analysis of thymocytes in thymus autonomy and turnover

Project description:Wild type thymi were transplanted into wild recipients (turnover) or Rag2-/-gc-/- mice (autonomy) and, after 28 days of transplant, thymocytes (double negative 3 and 4, immature CD8 single positive and CD4 CD8 double positive) were sorted and single cell RNAseq and TCR repertoire analysis performed

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The Zmiz1-Notch1 interaction induces Myc expression to drive steady state and stress thymopoiesis [DN3]

Project description:Notch1 signaling ramps up to very high levels in order to drive CD4-CD8- double-negative (DN) thymocytes to the CD4+CD8+ double-positive (DP) stage. During this important phase of T-cell development, which is known as the DN-DP transition, it is unclear whether the Notch1 complex simply strengthens its signal output as an isolated unit or recruits cofactors to amplify its signals. We previously showed that the PIAS-like coactivator Zmiz1 is a direct and context-dependent cofactor of Notch1 in T-cell leukemia. Using conditional knockout mouse models, we show that like inactivation of Notch, inactivation of Zmiz1 impaired the DN-DP transition under steady state and stress conditions. To determine mechanism, we performed RNA-Seq on sorted DN3 cells that were deprived of Zmiz1 signals either acutely (DeltaTamCre) or chronically (DeltaMxCre). To differentiate Notch1-independent from Notch1-dependent target genes, we also performed RNA-Seq on DN3 cells that were deprived of Notch1 signals using the anti-NRR Notch1 antibody. Our data suggests that Zmiz1 selectively amplifies a subset of Notch1 target genes in DN3 cells, such as Myc.

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Cellular heterogeneity of circulating CD4+CD8+ double positive T cells characterized by single-cell RNA sequencing

Project description:We subdivided circulating CD4+CD8+ double positive T cells, which are known to be related to various disease environments, into several subtypes through single-cell RNA sequencing, and compared gene signatures according to CD4 and CD8 protein expression levels using CD4 and CD8 antibody derived tags (feature barcode).

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The Zmiz1-Notch1 interaction induces Myc expression to drive steady state and stress thymopoiesis [ETP and DN3]

Project description:Notch1 signaling ramps up to very high levels in order to drive CD4-CD8- double-negative (DN) thymocytes to the CD4+CD8+ double-positive (DP) stage. During this important phase of T-cell development, which is known as the DN-DP transition, it is unclear whether the Notch1 complex simply strengthens its signal output as an isolated unit or recruits cofactors to amplify its signals. We previously showed that the PIAS-like coactivator Zmiz1 is a direct and context-dependent cofactor of Notch1 in T-cell leukemia. Using conditional knockout mouse models, we show that like inactivation of Notch, inactivation of Zmiz1 impaired the DN-DP transition under steady state and stress conditions. To determine mechanism, we performed RNA-Seq on sorted early T-lineage progenitor (ETP) and DN3 cells that were deprived of Zmiz1 signals either acutely (DeltaTamCre) or chronically (DeltaMxCre). To differentiate Notch1-independent from Notch1-dependent target genes, we also performed RNA-Seq on ETP and DN3 cells that were deprived of Notch1 signals using the anti-NRR Notch1 antibody. Our data suggests that Zmiz1 selectively amplifies a subset of Notch1 target genes in DN3 cells, such as Myc. In contrast, Zmiz1 does not have these functions in ETP cells, thus indicating stage-specific roles of Zmiz1.

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Project description:Mitochondria and endoplasmic reticulum contacts (MERCs) regulate multiple cellular processes including cell survival and differentiation. Based on the observations that MERCs were specifically enriched in the CD4-CD8- double negative (DN) stage, we studied their role in early thymocyte development. We found that T-cell-specific knockout of Hspa9, which encodes GRP75, a chaperone mediates MERC formation by assembling the IP3R-GRP75-VDAC complex, impaired DN3 thymocyte viability and resulted in thymocyte developmental arrest at the DN3-DN4 transition. Mechanistically, GRP75 deficiency induced mitochondrial stress, releasing mitochondrial DNA (mtDNA) into the cytosol and triggering the type I interferon (IFN-I) response. IFN-I pathway contributed to both the impairment of cell survival and DN3-DN4 transition blockage, while increased lipid peroxidation (LPO) played a major role downstream of IFN-I. Thus, our study reveals the essential role of GRP75-dependent MERCs in early thymocyte development and uncovers the governing facts of cellular survival and differentiation in the DN stage.

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Project description:T-cell-specific deletion of USP8 (USP8ffCD4Cre) revealed that USP8 is required for thymocyte transition to the CD4+ and CD8+ single positive (SP) stages. To evaluate underlying mechanisms, gene expression profilling was performed in CD4+CD8+ double pos thymocytes derived from control (USP8ff and USP8ffCD4Cre) mice. Microarray analysis of two groups of USP8fl/fl and USP8fl/flCD4Cre thymocytes. Material from two mice was combined for each group (eight mice in total).

2014-03-04 | E-GEOD-55517 | biostudies-arrayexpress

RNA-sequencing of mouse double-negative developing thymocytes [CD4-, CD8-, CD3-, B220-, CD25high, CD44low, and CD117low]

Project description:We performed a transcriptome comparison of double-negative developing thymocytes from the DN3 population, differing in their CD27 and CD28 expression

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Microarray analysis of gene expression in CD4+CD8+ double pos (DP) thymocytes upon depletion of Ubiquitin specific protease 8

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RNA-seq of Smarca5-deficient CD4/CD8 double-positive thymocytes isolated from 6-week mouse thymi and their comparison with wild-type and TP53-null controls.

Project description:Here we utilized a conditional knock-out mouse model to investigate the role of Smarca5, an ISWI subfamily chromatin remodeling ATPase, during thymocyte development using hCD2-iCre transgene. We did transcriptional profiling of FACS-sorted CD4/CD8 double-positive thymocytes as this thymic population was persistent yet strongly underrepresented in adult 6-week mutant thymi. Controls included age-matched CD4/CD8 double-positive thymocytes from wild-type and tumor suppressor protein Trp53-null mice. For comparison, the Smarca5/Trp53-double-deficient thymocytes included in the experiment were partially rescued upon loss of Trp53.

2019-04-01 | E-MTAB-7758 | biostudies-arrayexpress

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