Transcriptomic profiling of keratinocytes isolated from patients with rare inflammatory skin disorders
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ABSTRACT: Inflammatory skin diseases, including inflammatory linear verrucous epidermal naevus (ILVEN) and psoriasis, are known to collectively be hyperproliferative. We endeavoured to do a transcriptional comparative study on patient and control keratinocytes to uncover a final druggable common pathway for those disorders.
Project description:The methylation analysis of HPV+ HNSCC cell lines and HPV- HNSCC cell lines as well as clones from an infected HPV- cell line by Infinium HumanMethylation450 BeadChip Analysis of 24 450k methylation profiles, comprising 3 HPV+ HNSCC cell lines and 3 HPV- HNSCC cell lines (in duplicate each) and 12 infected HPV- HNSCC cell line clones
Project description:The life cycle of human papillomaviruses (HPV) is strictly linked to the differentiation of their natural host cells. The HPV E6 and E7 oncoproteins can delay the normal differentiation program of keratinocytes, however, the exact mechanisms responsible for this have not yet been identified. The goal of this study was to investigate the effects of HPV16 oncoproteins on the expression of genes involved in keratinocyte differentiation. Primary human keratinocytes transduced by LXSN (control) retroviruses or virus vectors expressing HPV16 E6, E7 or E6/E7 genes were subjected to gene expression profiling. The results of microarray analysis showed that HPV 16 E6 and E7 have the capacity to down-regulate the expression of several genes involved in keratinocyte differentiation. Quantitative real-time polymerase chain reaction (qRT-PCR) assays were performed to confirm microarray data. To investigate the effects of the HPV oncoproteins on the promoters of selected keratinocyte differentiation genes, luciferase reporter assays were performed. Our results suggest that the HPV 16 E6 and/or E7 oncogenes are able to down-regulate the expression of several genes involved in keratinocyte differentiation, at least partially by down-regulating their promoter activity. This activity of the HPV oncoproteins may have a role in the productive virus life cycle, and also in virus induced carcinogenesis. Primary human foreskin keratinocytes were transduced by retrovirus vectors containing HPV 16 E6, E7, E6/E7 or the control vector LXSN. The global gene expression patterns of transduced keratinocytes were analyzed on Affymetrix microarrays
Project description:Discover novel transcriptional units upstream of cell cycle genes Transcription of long noncoding RNAs (lncRNAs) within gene regulatory elements can modulate gene activity in response to external stimuli, but the scope and functions of such noncoding transcription are not known. Here we use an ultra-high density array that tiles the promoters of 56 cell cycle genes to interrogate 108 samples representing diverse perturbations. We identify 216 transcribed regions that encode putative lncRNAs--many of which have RT-PCR-validated periodic expression during the cell cycle, show altered expression in human cancers, and are regulated in expression by specific oncogenic stimuli, stem cell differentiation, or DNA damage. 53 samples tiled against respective controls
Project description:Human corneal endothelial cells (HCEC) form a monolayer by adhering tightly through their intercellular adhesion molecules. Located at the posterior corneal surface, they maintain corneal translucency by dehydrating the corneal stroma, mainly through the Na+- and K+-dependent ATPase (Na+/K+-ATPase). Because HCEC proliferative activity is low in vivo,we tried to activate proliferation of HCEC by inhibiting cyclin-dependent kinase inhibitors.We have here demonstrated microarray data of transduced human corneal endothelial cell lines. Affymetrix human U133 plus 2.0 array was used to transcriptionally profile to compare cultured human corneal endothelial cells and transduced human corneal endothelial cells.
Project description:Rabbit embryos, as in humans, develop as bilaminar discs at gastrulation and unlike egg cylinders as in rodents. Mammalian primordial germ cells (PGCs) in all species originate during gastrulation. We sequence the transcriptomes of rabbit embryos during gastrulation, and show that rabbit PGC (rbPGC) specification occurs at the posterior epiblast at the onset of gastrulation
Project description:Rabbit embryos, as in humans, develop as bilaminar discs at gastrulation and unlike egg cylinders as in rodents. Mammalian primordial germ cells (PGCs) in all species originate during gastrulation. We sequence the transcriptomes of rabbit embryos during gastrulation, and show that rabbit PGC (rbPGC) specification occurs at the posterior epiblast at the onset of gastrulation
Project description:Rabbit embryos, as in humans, develop as bilaminar discs at gastrulation and unlike egg cylinders as in rodents. Mammalian primordial germ cells (PGCs) in all species originate during gastrulation. We sequence the transcriptomes of rabbit embryos during gastrulation, and show that rabbit PGC (rbPGC) specification occurs at the posterior epiblast at the onset of gastrulation