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The transcription factor Gli3 promotes differentiation from double positive to CD4 single positive thymocyte by repression of Shh

ABSTRACT: We used Affymetrix microarrays to understand the genome wide differences in Wildtype and Gli3 mutant (Gli3+/- and Gli3-/-) (n=2) embryonic day 18.5 DP CD69-, DP CD69+ and SP4 thymocytes.

ORGANISM(S): Mus musculus

PROVIDER: GSE87499 | GEO | 2018/01/19

REPOSITORIES: GEO

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Expression data from CD4 single positive thymocyte subsets from C57BL/6 mice of 6-8 wks of age

Project description:After positive selection in the thymus, the newly generated single positive (SP) thymocytes are phenotypically and functionally immature and undergo apoptosis upon antigen stimulation. In the thymic medullary microenvironment, SP cells progressively acquire immunocompetence. Negative selection to remove autoreactive T cells also occur at this stage. We have defined four subsets of CD4 SP, namely, SP1, SP2, SP3, and SP4 that follow a functional maturation program and a sequential emergence during mouse ontogeny. We used microarray to detail the global programm of gene expression during the maturation of murine CD4 single positive thymocytes Four subsets of CD4+CD8-CD25-NK1.1- thymocytes from C57BL/6 mice of 6-8 wks of age were purified for RNA extraction and hybridization on Affymetrix microarrays, namely, SP1 (Qa-2-6C10+CD69+), SP2 (Qa-2-6C10-CD69+), SP3 (Qa-2-6C10-CD69-), SP4 (Qa-2+6C10-CD69-).

2012-02-13 | E-GEOD-30083 | biostudies-arrayexpress

Analysis of Differentially Expressed Genes in sorted CD69+ double positive (DP) thymocytes from wild type (WT) and NSRP1CD4Cre (KO) mice

Project description:NSrp70 deficiency (NSRP1f/fCD4Cre) profoundly perturbed the late development of DP thymocytes, leading to a significant reduction of single positive (SP) cells in the thymus and peripheral lymphoid tissues. To gain further insight into how NSrp70 controls thymocyte development from CD69+ DP stage, we performed RNA-seq analysis after sorting CD69+ DP thymocytes from WT and Nsrp1 cKO mice.

2020-06-27 | GSE153389 | GEO

Transcriptome of double-positive thymocyte precursors transduced with a retrovirus encoding Lin28a

Project description:We performed paired-end RNA-seq to compare the transcriptome of DP thymocytes that ectopically express Lin28 in vivo versus untransduced (GFP-ve) DP thymocytes.

2012-02-17 | GSE34854 | GEO

Transcriptome of double-positive thymocyte precursors transduced with a retrovirus encoding Lin28a

Project description:We performed paired-end RNA-seq to compare the transcriptome of DP thymocytes that ectopically express Lin28 in vivo versus untransduced (GFP-ve) DP thymocytes. Transduced (GFP+) and untransduced (GFP-) CD4+ CD8+ CD3- thymocytes were sorted and pooled from three recipients of hematopoietic stem and progenitor cells transduced with Lin28-RV six weeks post-reconstitution. Total RNA was extracted and paired-end library construction and sequencing was performed on oligo-dT purified RNA.

2012-02-17 | E-GEOD-34854 | biostudies-arrayexpress

Gli3 in fetal thymic epithelial cells promotes thymocyte positive selection and differentiation by repression of Shh.

Project description:Gli3 is a Hedgehog (Hh)-responsive transcription factor that can function as a transcriptional repressor or activator. We show that Gli3 activity in mouse thymic epithelial cells (TECs) promotes positive selection and differentiation from CD4+ CD8+ to CD4+ CD8- single-positive (SP4) cells in the fetal thymus and that Gli3 represses Shh Constitutive deletion of Gli3, and conditional deletion of Gli3 from TECs, reduced differentiation to SP4, whereas conditional deletion of Gli3 from thymocytes did not. Conditional deletion of Shh from TECs increased differentiation to SP4, and expression of Shh was upregulated in the Gli3-deficient thymus. Use of a transgenic Hh reporter showed that the Hh pathway was active in thymocytes, and increased in the Gli3-deficient fetal thymus. Neutralisation of endogenous Hh proteins in the Gli3-/- thymus restored SP4 differentiation, indicating that Gli3 in TECs promotes SP4 differentiation by repression of Shh Transcriptome analysis showed that Hh-mediated transcription was increased whereas TCR-mediated transcription was decreased in Gli3-/- thymocytes compared with wild type.

| S-EPMC5817998 | biostudies-literature

Expression data from CD4 single positive thymocyte subsets from C57BL/6 mice of 6-8 wks of age

2012-02-13 | GSE30083 | GEO

Transcriptome in naturally-occuring self-reactive thymocytes

Project description:In the Nur77/Nr4a1-eGFP BAC Tg reporter mice, GFP expression positively correlates with TCR signaling strength. We postulate that among post-selection thymocytes, GFP expression reflects self-antigen recognition across the TCR repertoire. Here we identify differentially expressed transcripts in semimature SP4 thymocytes (CD25- CD69+ CD24+) from Nur77-eGFP BAC Tg reporter mice sorted on 10% highest, intermediate, and lowest GFP expression.

2024-10-31 | GSE279344 | GEO

The transcription factor Gli3 promotes B cell development in the fetal liver through repression of Shh

Project description:Before birth B-cells develop in the fetal liver (FL). Here we show that Gli3 activity in the FL stroma is required for B-cell development. In the Gli3-deficient FL B-cell development was reduced at multiple stages, whereas the Sonic hedgehog (Shh)-deficient FL showed increased B-cell development, and Gli3 functioned to repress Shh transcription. Use of a transgenic Hedgehog (Hh)-reporter mouse showed that Shh signals directly to developing B-cells, and that Hh pathway activation was increased in developing B-cells from Gli3-deficient fetal liver. RNAsequencing confirmed that Hh-mediated transcription is increased in B-lineage cells from Gli3-deficient FL, and showed that these cells expressed reduced levels of B-lineage transcription factors and BCR/pre-BCR-signalling genes. We showed that expression of the master regulators of B-cell development, Ebf1 and Pax5, is reduced in developing B-cells from Gli3-deficient FL and increased in Shh-deficient FL, and that in vitro Shh-treatment or neutralisation can repress or induce their expression respectively.

2017-05-09 | GSE81467 | GEO

METTL3 governs thymocyte development and thymic involution by regulating ferroptosis

Project description:Given its central role in immune aging, identifying regulators of thymic involution is important. While conventional programmed cell death plays a fundamental role in thymocyte development, how cell death pathways contribute to thymic involution remains unclear. In this study, we found that CD4+CD8+ double positive (DP) thymocytes acquired characteristics of senescence in aged mice undergoing thymic involution, while expression of the m6A methyltransferase METTL3 (enriched in DP cells from young mice) decreased with aging. By conditionally deleting METTL3 in T cells, we revealed a critical role for METTL3 in DP cell survival, and in restraining features of aging in DP thymocytes by preventing ferroptosis signaling, through glutathione peroxidase 4 (GPX4). Mechanistically, GPX4 was maintained by METTL3 at the translational level, independent of its methyltransferase activity. Furthermore, we found that pharmacological inhibition of ferroptosis promoted DP cell survival, and attenuate d features of aging in DP thymocytes. These findings uncover a role for METTL3-regulated ferroptosis in thymic involution, and identify strategies to restore thymic function.

2024-08-18 | GSE271324 | GEO

Genes regulated by HEB and E2A during DP stage of thymocyte development

Project description:To investigate gene targets of the E-proteins HEB and E2A during the CD4+CD8+ double positive (DP) stage of T cell development. We examined E-protein function by simultaneous removal of both HEB (Tcf12) and E2A (Tcfe2a) genes at the DP stage. This was done by crossing mice containing HEB floxed and E2A floxed alleles to a CD4Cre background (Tcf12f/fTcfe2af/fCD4Cre mice). Microarray analysis was used to compare gene expression in HEB and E2A double deficient DP thymocytes (Cre+) to Cre- control DP thymocytes. Keywords: genetic modification

2007-12-04 | GSE9749 | GEO

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