Transcriptomic changes in lymphatic endothelial cells derived from psoriatic and healthy human skin
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ABSTRACT: Dermal lymphatic endothelial cells were isolated from chronic psoriatic plaques or healthy human skin for cultivation and subsequently subjected to RNA-Seq.
Project description:Dermal fibroblasts were isolated from healthy human skin or chronic psoriatic plaques for cultivation, which were subsequently subjected to RNA-Seq.
Project description:Single-cell transcriptomic profiling of CD45-CD31+ endothelial cells derived from healthy and psoriatic human skin using the 10x Genomics platform.
Project description:Alternative RNA splicing greatly increases proteome diversity, and the possibility of studying genome-wide alternative splicing (AS) events becomes available with the advent of high-throughput genomics tools devoted to this issue. Kaposi’s sarcoma associated herpesvirus (KSHV) is the etiological agent of KS, a tumor of lymphatic endothelial cell (LEC) lineage, but little is known about the AS variations induced by KSHV. We analyzed KSHV-controlled AS using high-density microarrays capable of detecting all exons in the human genome. Splicing variants and altered exon-intron usage in infected LEC were found, and these correlated with protein domain modification. The different 3’ UTR used in new transcripts also help isoforms to escape microRNA-mediated surveillance. Exome-level analysis further revealed information that cannot be disclosed using classical gene-level profiling: a significant exon usage difference existed between LEC and CD34+ precursor cells, and KSHV infection resulted in LEC-to-precursor, dedifferentiation-like exon level reprogramming. Our results demonstrate the application of exon arrays in systems biology research, and suggest the regulatory effects of AS in endothelial cells are far more complex than previously observed. This extra layer of molecular diversity helps to account for various aspects of endothelial biology, KSHV life cycle and disease pathogenesis that until now have been unexplored. 5 samples were analyzed. 3 were KSHV infected lymphatic endothelial cells (LECs), and 2 were non-infected control samples.
Project description:We microprepared native mammary skin resections from healthy female donors (breast size reduction) by a combination of enzymatic and mechanical treatment. The resulting suspensions of single dermal cells were then subjected to FACSorting using antibodies against the lymphovascular marker protein podoplanin and the panendothelial protein CD31 as positive and the leukocytic protein CD45 as negative markers. We aimed at separating lymphatic vascular endothelial cells (LECs) from blood vascular endothelial cells (BECs) in order to characterize their moelcular and functional phenotypes in health and disease. We found that lymphatic endothelial cells consisted of two instead of only one cell population. Their discrimination marker was high versus low expression of podoplanin surface protein, respectively. Especially, the low-podoplanin expressors were undescribed. Thus, we screened for their transcription profile using U133A. We identified specific marker genes and finally have assigned a specific function to the novel LEC subpopulation unknown up to now. Importantly, we did not use lysates from cell culture, but from ex vivo cells. Thus, there was no treatment of cells except processing the samples on ice.
Project description:Chromatin immunoprecipitation microarray (ChIP-chip) study using anti-Myc (9E10) antibodies and primary human lymphatic endothelial cells (LECs) transduced with recombinant adenoviruses expressing wild type or phosphorylation-deficient Myc-tagged FOXC2.
Project description:This SuperSeries is composed of the following subset Series: GSE16353: The profile of cellular and KSHV microRNAs in AIDS_KS biopsies (and normal skin control biopsies) GSE16354: Infection of Lymphatic and Blood Vessel Endothelial Cells (LEC and BEC) with KSHV GSE16355: Lymphatic endothelial cells (LEC) transfected with the KSHV microRNA cluster GSE16356: Lymphatic endothelial cells (LEC) treated with a MAF-targeted siRNA Refer to individual Series