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Genome-wide DNA methylation analysis on testicular embryonal carcinoma

ABSTRACT: Genomic DNA from 6 pure EC and 2 normal testis was fragmented and immunoprecipitated with anti-5mC monoclonal antibodies by MeDIP. After IP, DNA was purified and amplified using Whole Genome Amplification. Subsequently, DNA was biotin-labeled and hybridized to Human Tiling Array 2.0R Chips (Affymetrix) according to the manufacturer’s instruction. Raw data (CEL files) were normalized and analyzed by TAS (Affymetrix). Technical replicate of MeDIP-chip procedure was performed. Testicular embryonic carcinoma (EC) is a major subtype of non-seminomatous germ cell tumors (NSGCTs) in males. To identify epigenetic changes during testicular tumorigenesis, we profiled the DNA methylation of 6 ECs. These samples represent different stages (stage I and stage III) and different extent of invasiveness. Non-cancerous testicular tissues were included. A total of 1167 tumor-hypermethylated differential methylated regions (DMRs) were identified across the genome. Among them, 40 genes/ncRNA were found to have hypermethylated promoters. Interestingly, we found several hypermethylated sex-linked genes, including X-linked genes STAG2, SPANXD/E, MIR1184, Y-linked genes RBMY1A1/1B/1D and FAM197Y2P. Expression of RBMY1A was confirmed by immunohistochemistry in normal germ cells, but lost in EC and seminoma. Our genome-wide analysis identified methylation changes in several previously unknown genes for testicular ECs, which might provide insight into the crosstalk between normal germ cell development and carcinogenesis. A total of 8 DNA samples, including 6 EC and 2 normal testicular tissues

ORGANISM(S): Homo sapiens

SUBMITTER: Hoi-Hung Cheung

PROVIDER: E-GEOD-66784 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Genome-wide DNA methylation analysis on testicular embryonal carcinoma

Project description:Genomic DNA from 6 pure EC and 2 normal testis was fragmented and immunoprecipitated with anti-5mC monoclonal antibodies by MeDIP. After IP, DNA was purified and amplified using Whole Genome Amplification. Subsequently, DNA was biotin-labeled and hybridized to Human Tiling Array 2.0R Chips (Affymetrix) according to the manufacturer’s instruction. Raw data (CEL files) were normalized and analyzed by TAS (Affymetrix). Technical replicate of MeDIP-chip procedure was performed. Testicular embryonic carcinoma (EC) is a major subtype of non-seminomatous germ cell tumors (NSGCTs) in males. To identify epigenetic changes during testicular tumorigenesis, we profiled the DNA methylation of 6 ECs. These samples represent different stages (stage I and stage III) and different extent of invasiveness. Non-cancerous testicular tissues were included. A total of 1167 tumor-hypermethylated differential methylated regions (DMRs) were identified across the genome. Among them, 40 genes/ncRNA were found to have hypermethylated promoters. Interestingly, we found several hypermethylated sex-linked genes, including X-linked genes STAG2, SPANXD/E, MIR1184, Y-linked genes RBMY1A1/1B/1D and FAM197Y2P. Expression of RBMY1A was confirmed by immunohistochemistry in normal germ cells, but lost in EC and seminoma. Our genome-wide analysis identified methylation changes in several previously unknown genes for testicular ECs, which might provide insight into the crosstalk between normal germ cell development and carcinogenesis.

2015-03-12 | GSE66784 | GEO

Identification of differentially methylated regions between placentas and maternal blood cells on chromosome 18

Project description:Comparison of DNA methylation levels of chromosome 18 between placentas and blood cells of pregnant women Extracted DNA from 5 cases of placental tissues and 5 cases of maternal blood cell was subjected to MeDIP (methylated DNA immunoprecipitation) and hybridized to Affymetrix GeneChip® Human Tiling 2.0R Array sets. A control without immunoprecipitation (IP) was also processed in parallel with the immunoprecipitated DNA samples. The results were validated with bisulfite sequencing and mass spectrometry-based analysis

2010-12-22 | E-GEOD-22837 | biostudies-arrayexpress

Genome-wide analysis of differential methylation and gene expression in a testicular cancer cell line

Project description:Aberrant methylation has been postulated to play an important role in tumorigenesis. We report the use of methylated DNA immunoprecipitation (MeDIP) and whole-genome tiling arrays to investigate methylation changes in testicular germ cell tumor (TGCT) cells. Coupled to expression profiling changes, we found that only 22-26% of differentially methylated genes were also expressed differentially. This phenomenon was independent of the presence of CpG islands in the promoter. Differential methylation and expression of some of these genes were confirmed in testicular tumor tissue. A substantial number of differentially methylated regions in the human genome were not linked to annotated gene loci. Subsequent analysis indicated several microRNAs and small nucleolar RNAs were regulated by these differentially methylated regions. Our results demonstrate the power of the combination of MeDIP-chip analysis and expression profiling for discovery in cancer cells of epigenetically regulated genes and non-coding RNAs in cancer cells.

2009-03-18 | GSE15220 | GEO

Chromosome-wide DNA unmethylome profiling examining human chromosomes 2, 9, 19 and mouse chromosomes 1, 9, 19

Project description:The unmodified DNA fraction of jejunal enterocytes and the jejunum lacking enterocytes of humans and jejunal enterocytes of mice was interrogated on Affymetrix tiling arrays The mTAG technique was used to enrich the unmodified DNA fraction in a total of 56 humans and 26 mice

2016-03-25 | E-GEOD-69972 | biostudies-arrayexpress

MeDIP-chip of zdp mutant

Project description:Using meDIP-chip to provide methylation profile in zdp mutant and identify hypermethylated regions comparing to wild type Arabidopsis plants. Examination of zdp mutant's methylome. 4 samples(1 mutant, 1 WT, each has two biological replicates)

2012-03-20 | E-GEOD-33719 | biostudies-arrayexpress

Comparison of the testicular transcriptome of chromosomes 2, X, Y and the mitochondrial genome between two mouse species

Project description:We used tiling arrays to compare the testicular expression of chromosomes 2, X, Y and the mitochondrial genome between Mus m. domesticus and Mus spretus. We found an excess of dysregulated non-coding RNAs on chromosome 2. This group consists of intronic antisense RNAs, intergenic RNAs and also premature forms of PIWI-interacting RNAs (piRNAs). The unique transcriptional regulation in spermatogenesis is a characteristic of chromosome X. It is subject to meiotic transcriptional silencing, which is accompanied by generating X-chromosomal retrogenes on autosomes. We found striking dysregulation of the G6pdx - G6pd2 parental-retrogene pair between the mouse species. The postmeiotic expression of G6pdx in Mus spretus points out the continuous evolution of X chromosome silencing and provides clue for regulatory changes, which accompanies the out-of-X gene flow. Comparison between 3 samples of Mus spretus and 3 samples of Mus m. domesticus (C57BL/6J, B6). The supplementary file 'GSE19633_Spr_v_B6.bed' contains the chromosomal localizations of the probes together with logFC and adjusted P values.

2011-02-16 | E-GEOD-19633 | biostudies-arrayexpress

Unique and redundant function of SOX2 and SOX17 in regulating the germ cell tumour fate

Project description:Embryonal carcinomas (ECs) and seminomas are testicular germ cell tumours. ECs display expression of SOX2, while seminomas display expression of SOX17. In somatic differentiation, SOX17 drives endodermal cell fate. However, seminomas lack expression of endoderm markers, but show features of pluripotency. Here, we use ChIP-sequencing to report and compare the binding pattern of SOX17 in seminoma-like TCam-2 cells to SOX2 in EC-like 2102EP cells and SOX17 in somatic cells. In seminoma-like cells, SOX17 was detected at canonical (SOX2/OCT4), compressed (SOX17/OCT4) and other SOX family member motifs. SOX17 directly regulates TFAP2C, PRDM1 and PRDM14, thereby maintaining latent pluripotency and suppressing somatic differentiation. In contrast, in somatic cells canonical motifs are not bound by SOX17. In sum only 10% of SOX17 binding sites overlap in seminoma-like and somatic cells. This illustrates that binding site choice is highly dynamic and cell type specific. Deletion of SOX17 in seminoma-like cells resulted in loss of pluripotency, marked by a reduction of OCT4 protein level and loss of alkaline phosphatase activity. Further, we found that in EC-like cells SOX2 regulates pluripotency-associated genes, predominantly by partnering with OCT4. In conclusion, SOX17 (in seminomas) functionally replaces SOX2 (in ECs) to maintain expression of the pluripotency cluster.

2019-09-28 | GSE130519 | GEO

A single cell transcriptome atlas of murine endothelial cells

Project description:The heterogeneity of endothelial cells (ECs), lining blood vessels, across tissues remains incompletely inventoried. We constructed an atlas of >32,000 single-EC transcriptomic data from 11 tissues of the model organism Mus musculus. We propose a new classification of EC phenotypes based on transcriptome signatures and inferred putative biological features. We identified top-ranking markers for ECs from each tissue. ECs from different vascular beds (arteries, capillaries, veins, lymphatics) resembled each other across tissues, but only arterial, venous and lymphatic (not capillary) ECs shared markers, illustrating a greater heterogeneity of capillary ECs. We identified high-endothelial-venule and lacteal-like ECs in the intestines, and angiogenic ECs in healthy tissues. Metabolic transcriptomes of ECs differed amongst spleen, lung, liver, brain and testis, while being similar for kidney, heart, muscle and intestines. Within tissues, metabolic gene expression was heterogeneous amongst ECs from different vascular beds, altogether highlighting large EC heterogeneity.

2020-02-13 | E-MTAB-8077 | biostudies-arrayexpress

Endothelial cell heterogeneity in pathological angiogenesis characterized by metabolic transcriptome diversity

Project description:Endothelial cell (EC) metabolism is an emerging target for anti-angiogenic therapy in tumor and choroidal neovascularization (CNV), but little is known about individual EC metabolic transcrip-tomes. Here, by scRNA-sequencing 28,337 murine choroidal ECs (CECs) and sprouting CNV-ECs, we constructed a taxonomy to characterize their heterogeneity. Comparison with murine lung tumor ECs (TECs) revealed congruent marker gene expression by distinct EC phenotypes across tissues and diseases, suggesting similar angiogenic mechanisms. Trajectory inference of CNV-ECs revealed that differentiation of venous to angiogenic ECs was accompanied by metabolic transcriptome plasticity. EC phenotypes displayed metabolic transcriptome heterogeneity. Hypothesizing that conserved genes are more important, we used an integrated analysis, based on congruent transcriptome analysis, CEC-tailored genome scale metabolic modeling, and gene expression me-ta-analysis in multiple cross-species datasets, followed by functional validation, to identify the top-ranking metabolic targets SQLE and ALDH18A1, involved in EC proliferation and collagen production, respectively, as novel angiogenic targets.

2020-03-31 | E-MTAB-8604 | biostudies-arrayexpress

Transcriptome profiling of wild type and Pof mutant Drosophila melanogaster females

Project description:In Drosophila, two chromosome-wide compensatory systems have been characterized; the dosage compensation system acting on the male X-chromosome and the chromosome specific regulation of genes located on the heterochromatic 4th chromosome. Dosage compensation in Drosophila is accomplished by hypertranscription of the single male X-chromosome mediated by the MSL-complex. The mechanism for this compensation is suggested to be an MSL-complex mediated enhanced transcriptional elongation while the mechanism for the compensation mediated by Painting of fourth (POF) on the 4th chromosome has remained elusive. Here we show that POF binds to nascent RNA and this binding is associated with an increase in amount of chromosome 4 transcripts. Furthermore, genes located on the 4th chromosome are enriched in binding of the nucleoplasmic nucleporin component NUP98 and this enrichment correlates to increased POF binding. We also show that genes located in heterochromatic regions have a shorter transition time from site of transcription and to the nuclear envelope. Our current work broadens the understanding about how genes in heterochromatic regions can overcome the repressive influence of their hostile environment. Pof mutant vs. wild type, 3 replicates

2012-05-30 | E-GEOD-28513 | biostudies-arrayexpress

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