Quantitative Analysis of PPARD mRNA-Seq Transcriptomes of Mouse KRAS Mutant Pancreatic Tissues by Next Generation Sequencing (NGS)
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ABSTRACT:
PROVIDER: PRJNA735087 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR14729094_1.fastq.gz | Fastqsanger.gz | |||
| SRR14729094_2.fastq.gz | Fastqsanger.gz | |||
| SRR14729095_1.fastq.gz | Fastqsanger.gz | |||
| SRR14729095_2.fastq.gz | Fastqsanger.gz | |||
| SRR14729096_1.fastq.gz | Fastqsanger.gz |
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Project description:Purpose: NGS has revolutionized systems-based analysis of cell signaling pathways. The goal of this study is to determine the effects of PPARD in colon cancer cell transcriptomes in relation to the metastatic potential. Methods: NGS-derived colon cancer cell mRNA transcriptome profiles of HCT116 WT (HCT116) and HCT116 with genetic PPARD-knockout (KO1) cells were generated by deep sequencing, in quadruplicate, using Illumina HiSeq2000 .The transcriptomes of HCT116 and KO1 cells will be compared to determine the differentially expressed genes between HCT116 and KO1 cells. Differentially expressed genes will be examined in relation to the metastatic potential and validated by qRT-PCR. Results: Using an optimized data analysis workflow Tophat2, we mapped about 25 million sequence reads per sample to the human genome. Out of 22229 genes, we identified 12118 transcripts with >50 reads in at least one sample of HCT116 and KO1 cells with edgeR package and identified 6668 differentailly expressed genes with FDR 0.001 and P value cutoff 0.0022 using GLM tests fitted with BUM model. We further fltered the genes with both p-value and fold change and identified 416 genes with FDR 0.001 and fold change larger than 2. Among the differentially expressed genes, 311 were downregulated and 105 were upregulated in the KO1 cells compared with the WT cells. Twenty-three of the differentially expressed genes had significant association (i.e., a tendency towards co-occurrence) with PPARD expression (P < 0.05; log odds ratio > 1.5) in the TCGA colorectal adenocarcinoma database. Of these 23 genes, 7 were linked to metastasis by PubMed literature searches: GJA1, VIM, SPARC, NRG1, CXCL8 (IL-8), STC1, and SNCG, which were validated by q-RT-PCR. Conclusions: Our study represents the detailed analysis of PPARD transcriptomes in colon cancer cells, generated by mRNA-seq technology. Our results show that NGS offers a comprehensive and accurate quantitative and qualitative evaluations of mRNA contents in cells. We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biologic functions.
2016-12-01 | GSE89729 | GEO
Project description:Purpose: NGS has revolutionized systems-based analysis of cell signaling pathways. The goal of this study is to determine the effects of PPARD in gastric corpus epithenial cell transcriptomes in relation to gastric progenitor cell transfromation and gastric tumorigenesis. Methods: NGS-derived mRNA transcriptome profiles of gastric corpus epithelial cells from villin-PPARD and their age and sex matched WT mice at 10, 25 and 55 weeks were generated by deep sequencing (4-5 mice per group) using Illumina HiSeq4000 .The transcriptomes of villin-PPARD and WT mice were compared to determine the differentially expressed genes. Differentially expressed genes in the top canonical pathways will be examined and validated by qRT-PCR. Results: The raw data was aligned to the MM10 genome with Tophat2/2.0.14, and the number of reads per gene was counted with HTSeq/0.6.1.The read counts were normalized with "DESeq2". We used cutoff: FDR< 0.01 and fold change larger than 2 to identify 407 differentially expressed genes (DEGs) for 10 weeks, 717 DEGs for 25 weeks and 2694 DEGs for 55 weeks. Among those DEGs, 255 are shared by the 3 ages, in which 219 were upregulated and 36 were downregulated in villin-PPARD mice compared with the WT mice. Ingenuity Pathway Analysis for the 255 DGEs among the 3 age groups showed that IFN-g signaling was the top canonical pathway that PPAR-d activated, with an overlap ratio of 36.1% of the known genes involved in this pathway. The next two top canonical pathways, with overlap ratios of 23.7% and 15.9%, were the antigen presentation pathway and the pathway for activation of IRF by cytosolic pattern recognition receptors, respectively, both of which are related to IFN-g signaling pathway activation and immune inflammation. Conclusions: Our study represents the detailed analysis of PPARD transcriptomes in gastric corpus epithelial cells regulated by villin-promoter, generated by mRNA-seq technology. Our results show that NGS offers a comprehensive and accurate quantitative and qualitative evaluations of mRNA contents in cells. We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biologic functions.
2019-01-01 | GSE111050 | GEO
Project description:Pancreatic cancer is the most highly lethal disease with an increasing incidence. Accumulating evidence shows that as an initiating event of pancreatic tumorigenesis, mutant KRAS requires additional factors to promote pancreatic cancer progression. Peroxisome proliferator-activated receptor-delta/beta (PPAR-d) is a ligand-activating transcription factor that modulates many critical cellular functions and is upregulated in human pancreatic cancer tissues. To study PPAR-d's effects on mutant KRAS-initiated pancreatic tumorigenesis, we have generated a novel transgenic mouse model with simultaneously targeted PPAR-d overexpression and KrasG12D mutation (KRASmu) in pancreatic epithelial cells (called KC/Pd mice). Our preliminary data showed that PPAR-d ligand GW501516 strongly promoted pancreatic tumorigenesis in KRASmu mice (KC), and PPAR-d upregulation dramatically enhanced these effects in KC/Pd mice. However, molecular mechanisms by which PPAR-d and its synthetic ligand GW501516 promote KRASmu-initiated pancreatic tumorigenesis remains largely unknown. We have therefore performed genome-wide mRNA seq profiling analysis for pancreatic tissues from KC [WT] and KC/Pd (PD) mice fed GW501516 diet to discover the critical genes/pathways directly regulated by PPAR-d overactivation that accelerate pancreatic cancer progression. Pancreatic tissues of GW-treated KC/Pd (PD-GW) and KC (WT-GW) mice had distinctive differential expression patterns with 1498 differentially expressed genes (DEGs) according to a cut-off of p(Adj)<.05, including 709 upregulated DEGs and 789 downregulated DEGs. Furthermore, Gene set enrichment analyses of these 1498 DEGs using the ?Hallmark gene sets? category showed that the top enriched pathways included IL6-JAK-STAT3, inflammatory response, and KRAS signaling. Thus, the RNA-seq profiling analyses provide us important clues for molecular mechanisms by which PPAR-d overactivation promotes pancreatic cancer progression.
2022-03-02 | GSE176135 | GEO
Project description:Quantitative Analysis of PPARD RNA-Seq Transcriptomes of Mouse Gastric Corpus Epithenial Cells by Next Generation Sequencing (NGS)
| PRJNA435718 | ENA
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare lung organoids transcriptome profiling (RNA-seq) between wild type and KRAS mutation Methods: mRNA profiles of wild-type (WT) and KRAS mutated organoids were generated by deep sequencing, in triplicate, using Illumina HiSeq. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks.
2023-06-01 | GSE171728 | GEO
Project description:The PPAR (Peroxisome proliferator-activated receptor) family of nuclear receptors has three members: PPARg, PPARa and PPARd. Although they share similar structures, their biological functions are distinct. PPARg controls lipid storage and adipogenesis, while PPARd is associated with fat burning. The highly specific synthetic ligand for PPARd, GW501516, is a promising drug candidate for obesity and diabetes. Here we use Affymetrix microarray to analyze gene expression profile in mouse embryo fibroblasts treated with 100 nM GW501516 for 0, 2, 8 and 24 hours. These data may provide new clues into the molecular mechanism by which GW501516 ameliorates obesity and diabetes.
2013-12-31 | GSE20371 | GEO
Project description:The PPAR (Peroxisome proliferator-activated receptor) family of nuclear receptors has three members: PPARg, PPARa and PPARd. Although they share similar structures, their biological functions are distinct. PPARg controls lipid storage and adipogenesis, while PPARd is associated with fat burning. The highly specific synthetic ligand for PPARd, GW501516, is a promising drug candidate for obesity and diabetes. Here we use Affymetrix microarray to analyze gene expression profile in mouse embryo fibroblasts treated with 100 nM GW501516 for 0, 2, 8 and 24 hours. These data may provide new clues into the molecular mechanism by which GW501516 ameliorates obesity and diabetes. Wild type mouse embryonic fibroblasts (MEFs) stably infected with retroviruses MSCVpuro expressing PPARd were plated at 0.5 million per 10 cm dish. After overnight incubation, cells were treated with 100nM GW501516 for 0h, 2h, 8h and 48h. Cells were collected at subconfluent condition. Total RNAs were sequentially purified with Trizol (Invitrogen) and RNeasy kit (Qiagen) and analyzed in triplicate on Mouse Genome 430 2.0 Array (Affymetrix) at NIDDK Microarray Core Facility following standard protocols.
2013-12-31 | E-GEOD-20371 | biostudies-arrayexpress
Project description:Using NGS approach we performed the search of multiple sclerosis-related miRNAs. We used PBMC as an informative and easily accessible biological material. To exclude bias in miRNA expression levels caused by disease modifying therapies, miRNA profiling was performed in treatment-naïve RRMS patients. Taking into account hypothetic gender specificity in disease pathogenesis we compared miRNA expression in RRMS patients and HCs separately for men and women. MiRNA profiling in men identified 32 differentially expressed miRNAs, which passed threshold for multiple corrections and may be attributed to MS-related. At the same time we did not find well-defined MS-specific miRNA expression signatures in women using NGS
2019-01-11 | GSE124900 | GEO
Project description:The role of murine peroxisome proliferator-activated receptor-delta (PPARd) in mammary tumorigenesis was assessed. Microarrays were used to analyse global gene expression to determine changes in MMTV-PPARd transgenic mice versus wild-type mice and the effect of GW501516.
2013-05-31 | GSE34109 | GEO