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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Gene Expression Profile of Medullary Epithelial Cells isolated from thymus of Bone Marrow (BM) reconstituted RAG1 null (control) and SCID NOD mouse

ABSTRACT: We identified DNAPK as one of the major proteins that physically interact with Autoimmune regulator (Aire). To establish physiological significance of DNAPK in Aire-driven expression of PTA genes in MECs, we utilized BM-reconstituted SCID mice (which express non functional DNAPK in their MECs) and RAG1 null mouse as a control. MECs from these mutated/reconstituted mice were sorted by Flow cytomety and their expression profile was analyzed using Affymetrix chips. The analysis shows that Aire-dependent PTA genes are significantly underexpressed in reconstituted SCIDS vs. RAGs (Controls), suggesting the involvement of DNAPK in Aire-mediated gene expression. Keywords: gene mutation Since SCID mice lack mature thymocytes and as a result thymic stroma, we reconstituted these animals by BM transfer (IP injection into 2-4d pups) 6 hrs post-irradiation (200rad). Reconstituted thymi were analyzed 6-8w after transfer.

ORGANISM(S): Mus musculus

SUBMITTER: Christophe Benoist

PROVIDER: E-GEOD-24734 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Gene Expression Profile of Medullary Epithelial Cells isolated from thymus of Bone Marrow (BM) reconstituted RAG1 null (control) and SCID NOD mouse

Project description:We identified DNAPK as one of the major proteins that physically interact with Autoimmune regulator (Aire). To establish physiological significance of DNAPK in Aire-driven expression of PTA genes in MECs, we utilized BM-reconstituted SCID mice (which express non functional DNAPK in their MECs) and RAG1 null mouse as a control. MECs from these mutated/reconstituted mice were sorted by Flow cytomety and their expression profile was analyzed using Affymetrix chips. The analysis shows that Aire-dependent PTA genes are significantly underexpressed in reconstituted SCIDS vs. RAGs (Controls), suggesting the involvement of DNAPK in Aire-mediated gene expression. Keywords: gene mutation

2010-10-16 | GSE24734 | GEO

Expression data from WT and Aire KO thymic medullary epithelial cells (MECs)

Project description:The aim of this study is to analyze the transcriptional effects of Aire deficiency in the thymus, using the Affymetrix MoGene platform to analyze variation in exon usage MECs were isolated from 4-6 wk-old WT or Aire KO ((B6xNOD)F1 background) mice. Three WT and three Aire-KO mice taken individually were used.

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2022-08-12 | PXD029867 | Pride

Gene expression profiling of perinatal or adult medullary thymic epithelial cells (MECs)

Project description:A comparative analysis of gene expression of perinate or adult Aire-GFP+ and GFP- MECs in WT or Aire-KO thymus Aire+ and Aire- MECs were sorted from adults (5 weeks old) or perinates (0-3 days old) of Aire-driven Igrp-GFP (Adig) mice in Aire-WT and -KO background. RNA from whole samples was amplified, labeled, and hybridized to Affymetrix Mouse Gene 1.0 ST Arrays.

2015-03-02 | E-GEOD-66402 | biostudies-arrayexpress

Differential features of Aire-induced and Aire-independent promiscuous gene expression in thymic epithelial cells

Project description:Promiscuous gene expression (pGE) of numerous self-antigens in thymic epithelial cells (TEC) enables the elimination of self-reactive T cells. The autoimmune regulator (Aire) is the only known molecular determinant driving pGE in the thymus but the existence of Aire-independent mechanisms has been inferred. Here, we analyzed the poly(A)+ transcriptome of TEC populations by RNA-sequencing (RNA-seq) in order to reveal differential features of Aire-induced vs. –independent pGE. We report an unanticipated effect of Aire deletion on the proliferation and differentiation of cortical TEC. Moreover, the RNA-seq data reveal the breath of Aire-induced and –independent pGE in medullary TEC (mTEC) subsets and the extent of thymic peripheral tissue representation. The results suggest that Aire-induced promiscuously expressed transcripts affect several functions with far reaching biological consequences in mTEC. High-throughput characterization of TEC transcriptomes will enable progress in understanding TEC biology and the establishment of self-tolerance. The mRNA profiles of cTEC, mTEClo and mTEChi from 6-8 week-old wild type (WT) and Aire-/- (KO) mice were generated by RNA-sequencing using Illumina HiSeq2000.

2015-05-19 | E-GEOD-65617 | biostudies-arrayexpress

Gene expression profiling of perinatal or adult medullary thymic epithelial cells (MECs)

Project description:A comparative analysis of gene expression of perinate or adult Aire-GFP+ and GFP- MECs in WT or Aire-KO thymus

2015-03-02 | GSE66402 | GEO

Aire, guardian of immunological tolerance, binds to and activates super-enhancers [ChIP-seq]

Project description:ChIP-seq profile of Aire and partners in MECs

2017-01-22 | GSE92597 | GEO

The effect of the autoimmune regulator (Aire) gene on the transcriptional expression profile of 3.10 mTEC cell line.

Project description:Medullary thymic epithelial cells (mTECs) contribute to self-tolerance through the ectopic expression of peripheral tissue antigens (PTAs) in the thymus. PTA expression in mTECs is largely dependent on the autoimmune regulator (Aire) gene. Here we used a Mus musculus mTEC cell line (3.10 mTEC line, which constitutively express Aire in culture) to knockdown Aire gene by means of siRNA transfection. Aire knockdown was confirmed by means of qRT-PCR and RNA-FISH (for Aire mRNA levels), immunofluorescence and western blot (for AIRE protein levels).The Agilent oligo microarrays were used to determine the large scale transcriptional expression profiles of control or Aire-knockdown 3.10 mTECs.

2015-12-31 | E-MTAB-3879 | biostudies-arrayexpress

Identification of a novel Aire-regulating cis-regulatory element using H3K27ac ChIP-seq

Project description:Thymic central tolerance is essential to preventing autoimmunity. In medullary thymic epithelial cells (mTECs), the Autoimmune regulator (Aire) gene plays an essential role in this process by driving the expression of a diverse set of tissue-specific antigens (TSAs), which are presented and help tolerize self-reactive thymocytes. Interestingly, Aire has a highly tissue-restricted pattern of expression, with only mTECs and peripheral extrathymic Aire-expressing cells (eTACs) known to express detectable levels in adults. Despite this high level of tissue specificity, the cis-regulatory elements that control Aire expression have remained obscure. We used sequence conservation analysis and ChIP-seq against the enhancer-associated histone mark H3K27ac to identify a candidate Aire cis-regulatory element. There is enrichment of H3K27ac near this element, ACNS1, in mTECs and the element also has characteristics of being NF-ÎºB-responsive. Finally, we find that this element is essential for Aire expression in vivo and necessary to prevent spontaneous autoimmunity, reflecting the importance of this regulatory DNA element in promoting immune tolerance. Two experimental groups (GFP neg mTECs and GFP pos mTECs), each with three samples, and one control sample (D10 Th2 cells).

2015-10-23 | E-GEOD-74257 | biostudies-arrayexpress

The effect of the Autoimmune regulator (Aire) gene on microRNA transcriptional profile of 3.10 mTEC cell line

Project description:Medullary thymic epithelial cells (mTECs) contribute to self-tolerance through the ectopic expression of peripheral tissue antigens (PTAs) in the thymus. Autoimmune regulator (Aire) is to date the best characterized transcriptional regulator known to at least partially coordinate PTA expression in mTECs. Furthermore the expression of Aire-dependent and -independent genes is also modulated by microRNAs. Given the transcriptional regulation exerted by Aire in mTEC cells and the role of miRNAs in post-transcriptional control, we used a Mus musculus mTEC cell line (3.10) which constitutively express Aire in culture) to knockdown Aire gene by means of siRNA transfection and then observe the effect of Aire knockdown in the transcriptional profile of miRNAs by means of oligomicroarrays. The Agilent oligomicroarrays were used to determine the large scale the miRNA transcriptional profiles of control or Aire-knockdown 3.10 mTECs.

2016-06-27 | E-MTAB-4596 | biostudies-arrayexpress

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