Project description:In this study, mRNA expression profiles were examined by Illumina microarray in mouse embryonic stem cells (ESCs) derived from androgenetic (aESC), parthenogenetic (pESC) and fertilized (fESC) blastocysts. Results showed that 2394, 87 and 1788 mRNAs were differentially expressed in the aESCs vs. fESCs, pESCs vs. fESCs and aESCs vs. pESCs, respectively. Androgenetic, parthenogenetic and fertilized embryonic stem cell lines were established from androgenetic, parthenogenetically activated and fertilized blasotocyst. mRNA microarrays were repeated three times using passages 6, 7 and 8.
Project description:In this study, miRNA expression profiles were examined by Illumina microarray in mouse embryonic stem cells (ESCs) derived from androgenetic (aESC), parthenogenetic (pESC) and fertilized (fESC) blastocysts. Results showed that 125, 42 and 99 miRNAs were differentially expressed in the aESCs vs. fESCs, pESCs vs. fESCs and aESCs vs. pESCs, respectively. Androgenetic, parthenogenetically activated and fertilized embryonic stem cell lines were established from androgenetic and fertilized blasotocyst. microRNA microarrays were repeated three times using passages 6, 7 and 8.
Project description:Measurement of specific gene expression in clinical samples is a promising approach for monitoring the recipient immune status to the graft in organ transplantation. Identification of biomarker genes closely associated with tolerance or rejection is critical for this monitoring protocol. Unlike previous studies, our microarray analysis focused on donor antigen-reactive T cells, which were prepared by collecting CD69+ T cells from cocultures of recipient peripheral T cells and donor antigen-presenting cells. A comparison of different recipient groups enabled us to identify several tolerance- and rejection-correlated biomarker genes, including previously unknown genes. By measuring biomarker gene expression in the CD69+ T cell fraction using quantitative reverse-transcription polymerase chain reaction, we were able to precisely detect the immune status of recipients relative to their graft. Full-thickness donor (BDF1; C57BL/6 x DBA/2) tail skin was grafted onto recipient (C57BL/6) mice. Tolerance to the graft was induced by donor specific transfusion combined with administration of anti-CD40L Ab (MR-1). Total T lymphocytes were prepared from spleen and lymph nodes of tolerant recipient mice on day 21 and 100 after skin transplantation, untreated recipient mice on day 7, and ungrafted, healthy C57BL/6 mice and used as early tolerant, tolerant, rejecting and naïve T lymphocyte samples, respectively. In order to isolate alloantigen-reactive cells, T lymphocytes were cocultured with donor BM-DCs for 20-21 hours and then separated into two subpopulations based on CD69 expression using fluorescence cell sorter. Total RNA was extracted from these CD69+ and CD69- T cells and subjected to microarray using Illunima MouseRef-8 Expression BeadChip. All data analysis and visualization of differentially expressed genes was conducted using Illumina BeadStudio v3.1.3 (Gene Expression Module v3.3.8), Array Assist 5.5.1 (Stratagene) and R 2.4.1(www.r-project.org). Candidate tolerance genes were defined as the common genes satisfying the following criteria; 1) 1.5 fold or higher expression in CD69+ T cells isolated from tolerant recipients than those from naïve mice, 2) 1.5 fold or higher expression in CD69+ T cells than CD69- T cells from tolerant mice, 3) 500 or higher average signal measured in CD69+ T cells from tolerant mice. The criteria for candidate rejection genes were 1) 1.5 fold or higher expression in CD69+ T cells isolated from rejecting recipients than those from tolerant mice, 2) 1.5 fold or higher expression in CD69+ T cells than CD69- T cells from rejecting mice, 3) 500 or higher average signal measured in CD69+ T cells from rejecting mice.
Project description:Rationale: COPD is characterized by an abnormal regulatory T cell (Treg) response and increases in Th1 and Th17 cell responses. It is unclear if dysregulation of miRNAs within Treg alters the adaptive immunophenotype in COPD. Objectives: To compare the miRNA profile of COPD Treg cells with that of healthy controls and to explore the function of differentially expressed miRNAs. Methods: Treg cells (CD4+CD25+CD49-CD127-) and T effector (CD4+CD25-) cells were obtained from the peripheral blood of 4 nonsmokers, 4 healthy current smokers, and 4 COPD current smokers for analysis of miRNA expression, matching them for age and sex. We assessed expression initially by microarray analysis on the Illumina miRNA platform then conducted real time RT-PCR validation of the microarray results. 24 samples were analyzed. 8 from each patient group of healthy Normal, Healthy Smoker, and COPD Smoker.
Project description:In order to investigate miRNA alterations associated to early relapse in ovarian cancer patietns, we analyzed miRNA expression profile in a test set of 30 surgical specimens including 13 early and 17 late relapsing patients. Samples included in test set were obtained from formalin-fixed paraffin-embedded (FFPE) specimens. Patients were selected on the basis of residual disease after primary surgery and time to relapse (TTR) after front-line chemotherapy. For the test set, a selection of the outliers concerning TTR was made: 12 months from the end of therapy was the TTR selected to define early (<12 months) or late (>12 months) relapsing patients. Clinical codes: Histotype: according to International Federation of Gynecological and Obstetrics guidelines Stage: according to International Federation of Gynecological and Obstetrics guidelines Grading: according to International Federation of Gynecological and Obstetrics guidelines Debulking: NED: not evident disease; mRD: minimal residual disease; GRD: gross residual disease Therapy code: P: Platinum without taxanes; PT: Platinum/paclitaxel End Point: Early: relapse whithin 12 and 6 months from the end of therapy for optimally (NED+ mRD) and sub-optimally (GRD) debulked patients respectively. Late: median time to relapse 48 and 34 months from the end of therapy for optimally and sub-optimally debulked patients respectively.
Project description:In order to investigate miRNA alterations associated to early relapse in ovarian cancer patients, we analyzed miRNA expression profile in a training set of 55 surgical specimens including 30 early relapsing and 25 late relapsing patients. Patients were selected on the basis of residual disease after primary surgery and time to relapse (TTR) after front-line chemotherapy. For training set, a selection of the outliers concerning TTR was made: as early relapsing were defined optimally debulked (OD) patients with a TTR< 12 months and sub-optimally debulked (SOD) patients with TTR< 6 months; late relapsing were defined OD patients with TTR> 36 months and SOD patients with TTR> 12 months. Clinical codes: Histotype: according to International Federation of Gynecological and Obstetrics guidelines Stage: according to International Federation of Gynecological and Obstetrics guidelines Grading: according to International Federation of Gynecological and Obstetrics guidelines Debulking: NED: not evident disease; mRD: minimal residual disease; GRD: gross residual disease Therapy code: P: Platinum without taxanes; PT: Platinum/paclitaxel End Point: Early: relapse whithin 12 and 6 months from the end of therapy for optimally (NED+ mRD) and sub-optimally (GRD) debulked patients respectively. Late: median time to relapse 48 and 34 months from the end of therapy for optimally and sub-optimally debulked patients respectively.
Project description:This SuperSeries is composed of the following subset Series: GSE32952: Gene expression profiles of T cell fraction enriched for donor antigen-reactive T cells in skin allografted recipient mice (I) GSE32953: Gene expression profiles of T cell fraction enriched for donor antigen-reactive T cells in skin allografted recipient mice (II) Refer to individual Series
Project description:The first round of translation occurs on mRNAs bound by nuclear cap-binding complex (CBC), which is composed of nuclear cap-binding protein (CBP) 80 and 20. During this round of translation, aberrant mRNAs are recognized and downregulated in abundance by nonsense-mediated mRNA decay (NMD), which is one of the mRNA quality control mechanisms. Here our microarray analysis reveals that the level of cyclin-dependent kinase inhibitor 1A (CDKN1A) mRNAs increases in the cells depleted of cellular NMD factors. Intriguingly, CDKN1A mRNA contains an upstream open reading frame (uORF), which is one of NMD-inducing features. Using chimeric reporter constructs and confocal microscopy, we find that the uORF of CDKN1A mRNA is actively translated and modulates a translational efficiency of the main downstream ORF. Our findings provide the biological insights into the possible role of NMD in diverse pathways mediated by CDKN1A. The microarray analysis performed to analize the cellular NMD substrates that regulated by Upf1, PNRC2 and/or CTIF in HeLa cell. The hypothesis tested in the present study was that endogenous NMD substrates may co-regulated by Upf1, PNRC2 and CTIF. Results provide important information that vast range of cellular NMD substrates are reqired CTIF. Total RNA obtained from HeLa cells with downregulation of Upf1, PNRC2 or CTIF by siRNA. The up- or down-regulated transcripts were compare to control siRNA treated HeLa cell RNA extract. Significant transcripts were confirmed by replication.
Project description:Analysis of cellular SMD or NMD substrates that regulated by Upf1 and/or PNRC2 in HeLa cell. The hypothesis tested in the present study was that endogenous SMD or NMD substrates may co-regulated by Upf1 and PNRC2. Results provide important information that vast range of cellular SMD or NMD substrates are reqired PNRC2 for decay. Total RNA obtained from HeLa cells with downregulation of Upf1 or PNRC2 by siRNA. The up- or down-regulated transcripts were compare to control siRNA treated HeLa cell RNA extract. Significant transcripts were confirmed by replication
Project description:This SuperSeries is composed of the following subset Series: GSE25202: microRNA alterations associated to clinical response in advanced stage ovarian cancer patients (training set). GSE25203: microRNA alterations associated to clinical response in advanced stage ovarian cancer patients (test set). Refer to individual Series