Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling time series of thymocytes from HNT-TCR transgenic mice injected with HA peptide to induce thymic negative selection

ABSTRACT: During T cell development in the thymus, negative selection eliminates autoreactive and potentially harmful T cells, leading to central tolerance. This process, involves specific apoptosis programs thought to be governed by the transcription factor Nur77. To analyze the genetic pathways involved in thymic negative selection and to point to new genes involved in thymic selection, we performed a high throughput temporal expression profiling of thymocytes from HNT-TCR transgenic mice undergoing specific peptide-mediated thymic negative selection, using oligonucleotide microarrays. We used HNT-TCR transgenic mice that are on a B10D2 background and express a transgenic TCR (V beta 8), specific for the HA peptide (HNTNGVTAACSHE) presented by MHC class II I-A^d. In this model of thymic negative selection, a single injection of the specific HA peptide induces massive and synchronized deletion of thymocytes. In this study, we analyzed the dynamically regulated genes over a 24 hours period, with a focus on the early time points of 3 and 6 hours after peptide injection.

ORGANISM(S): Mus musculus

SUBMITTER: unknown ripoche

PROVIDER: E-TABM-550 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Context : Non Functioning Pituitary Adenomas (NFPAs), although typically benign, may be locally invasive. Few datas are currently available related to the molecular process of invasion in sporadic pituitary adenomas. Markers of invasiveness, important for helping the clinician in the therapeutic strategy particularly in the decision for adjuvant radiotherapy, are currently lacking. Objective: Evaluate if invasive NFPAs display a specific expression profile compared with non invasive tumors. Methods: To address this issue, we selected 40 NFPAs (38 of the gonadotroph type) and classified them as invasive (n=22) or non invasive (n=18) on the basis of MRI and surgical findings. Then we performed pangenomic analysis based on Agilent Human Whole genome Gene Expression oligonucleotide microarray (44k) Expression in order to identify genes differentially expressed between invasive and non invasive NFPA. Experiements are made in dual color with tumor samples labelled in cyanine 5 and a pool of all tumors labelled in cyanine 3.The expressions of some genes identified by microarray screening were confirmed by real-time quantitative RT-PCR. The selection of genes was made on the basis of Ingenuity networks and degree of upregulation between invasive and non invasive tumors. Moreover, some genes of interest already described in the literature were added to the analysis. Results: Prediction class analysis showed that 346 genes discriminated between invasive and non invasive NFPAs (p<0.001); 233 were up- and 113 were down-regulated between the two groups. We then tested On the basis of Ingenuity networks and degree of upregulation between invasive and non invasive tumors, a set of eight genes, including MYO5A, IGFBP5, FLT3, NFE2L1, PTTG, MMP9, NCAM and NR1H3, was tested in quantitative PCR analysis and. Results confirmed those obtained in microarrays results.analysis. At the protein level, Myosin 5A (MYO5A) demonstrated stronger immunostaining in invasive NFPAs compared to non invasive NFPAs. Conclusions: We propose a molecular signature of eight genes of grossly invasive NFPAs as compared with non invasive tumors: The product of one of these genes, MYO5A, may be a useful marker of invasive process in tumoral specimens. The role of these genes in the invasiveness process and as prognostic markers of pituitary adenomas needs to be confirmedinvestigated. Method: Microarray analyses the transcriptome of 22 invasive Non Functional Pituitary Adenomas (NFPAs) and 18 non invasive NFPAs in dual color (each sample labelled in cyanine 5 and a pool of all tumors labelled in cyanine 3).

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Transcription profiling time series of thymocytes from HNT-TCR transgenic mice injected with HA peptide to induce thymic negative selection

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