KSHV G-protein-coupled receptor (GPCR) induced gene transcription in HUVEC cells
Ontology highlight
ABSTRACT: We used microarrays to compare the gene transcription level between HUVEC-kGPCR and HUVEC-vector stable cells. The goal is to identify the genes that are affected by KSHV kGPCR expression. We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of up-regulated genes during this process.
Project description:We used microarrays to compare the gene transcription level between HUVEC-kGPCR and HUVEC-vector stable cells. The goal is to identify the genes that are affected by KSHV kGPCR expression. We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of up-regulated genes during this process. HUVEC cells were infected with kGPCR or control lentivirus and selected with puromycin to get stable cells. Then RNA was extracted from these cells and microarray analysis was performed.
Project description:Expression profiling of latently infected cells using a custom tiling microarray HUVEC and TIME cells were infected BCBL-1-derived KSHV. Mock infected HUVEC and TIME cells served as controls for each of these two stably infected cells, respectively. BJAB cells served as uninfected controls for the BCBL-1 cells. KSHV-infected cells are induced to enter lytic cycle with valproate or Adenovirus-RTA. Cells were harvested at indicated time points and analyzed.
Project description:Expression profiling of latently infected cells using a custom tiling microarray HUVEC and TIME cells were infected BCBL-1-derived KSHV. Mock infected HUVEC and TIME cells served as controls for each of these two stably infected cells, respectively. BJAB cells served as uninfected controls for the BCBL-1 cells. KSHV-infected cells are induced to enter lytic cycle with valproate or Adenovirus-RTA. Cells were harvested at indicated time points and analyzed. Three condition experiment: mock infected, latently infected cells and lytically infected. Three cell types (BJAB cells served as uninfected controls for the BCBL-1 cells).
Project description:The purpose was to measure expression changes in human circular RNAs. Total RNA was harvested from KSHV-infected HUVEC and MC116 cells. A portion of the RNA was digested with RNase R to enrich for circular RNAs.
Project description:Kaposi's sarcoma-associated herpesvirus (KSHV) is the causative agent of Kaposi's sarcoma (KS) and two human lymphoproliferative diseases: primary effusion lymphoma and AIDS-related multicentric Castleman's disease. KSHV latency-associated nuclear antigen (LANA) is expressed in KSHV-infected cancer cells and is responsible for maintaining viral genomes in infected cells. Thus, LANA is an attractive target for therapeutic intervention for KSHV-associated diseases. Here, we devised a gene therapy vector using the adeno-associated virus (AAV), which capitalizes the LANA's function to tether terminal repeat (TR) containing circular genome in latently infected cells and the TR's enhancer function for KSHV inducible gene promoters. By including two TR copies with a lytic inducible gene promoter (TR2-Orip), we generated an AAV vector, which expresses an engineered thymidine kinase (TK) selectively in KSHV-infected cells. Ganciclovir (GCV), an anti-herpesvirus drug, effectively eradicated multiple KSHV-infected cells that include iPSC derived epithelial colony forming cells, but not non-KSHV-infected counterparts in the presence of AAV8-TR2-Orip-TK. In addition, AAV8-TR2-Orip-TK prevents KSHV virion production from reactivated cells, hence spreading KSHV infections from reactivated cells. Anti-cancer drugs, known to reactivate KSHV, stimulated TK expression from the vector and, therefore, synergized with AAV8 TR2-Orip-TK to induce KSHV-infected cancer cell death. Finally, the AAV8-TR2-Orip-TK with GCV completely diminished KSHV-infected cancer cells in the xenograft tumor model. Our new cancer gene therapy should augment the current clinical protocol for KS.
Project description:By constructing specific lentiviral vector, vefg165 protein was overexpressed in HUVEC cells. Using light-curing bioprinting, HUVEC(vefg165+)-loaded hydrogels and HUVEC(vector)-loaded hydrogels were prepared, and were used to treat rat diabetic wounds. The regenerated skin tissues (five samples for each group) were analyzed with high-throughput proteomics.
Project description:Kaposi sarcoma is the most common cancer in AIDS patients and is typified by red skin lesions. The disease is caused by the KSHV virus (HHV8) and is recognisable by its distinctive red skin lesions. The lesions are KSHV-infected spindle cells, most commonly the lymphatic endothelial and blood vessel endothelial cells (LEC and BEC), plus surrounding stroma. The KSHV virus expresses multiple microRNA in a single cluster. Here we test the effects of this KSHV microRNA cluster in LEC cells using Affymetrix hgu133plus2 chips. Experiment Overall Design: There are n=3 of: 1. LEC control with empty vector pSIN-MCS (LEC), 2. LEC transfected with the same vector containing the KSHV microRNA cluster.
Project description:Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a viral RNA-binding protein ORF57 that plays an essential role in posttranscriptional regulation of viral gene expression. Ectopice expression of KSHV ORF57 in HEK293T cells was evaluated on its effect on host gene expression in the study, with the cells transfected with an empty vector serving as a control.
Project description:We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of up-regulated genes during this process.
Project description:We used ac4C-seq to detedc cellular and KSHV transcripts in iSLK-Puro, iSLK-KSHV, iSLK-KSHV (WT), iSLK-KSHV (ΔNAT10) cells after after doxycycline and sodium butyrate treatment.