RNA-seq of human Caco-2 cells treated with a nutraceutical formula (solution-3), or vehicle-(saline solution)- control, and infected with SARS-CoV-2 (EG.5)
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ABSTRACT: Caco-2 cells were plated (5x105 cells), treated with a nutraceutical formula (solution-3) at 0.01x concentration or with vehicle (saline solution) as the negative control for the treatment. After 1 hour, the cells were infected with SARS-CoV-2 viral particles (EG.5; MOI, 3). After 48 hours, the cells were lysed, and their RNA were extracted for RNAseq.
Project description:Caco-2 cells were plated (1000000 cells) and treated with a nutraceutical formula (Solution-3) at 0.01x concentration or with vehicle (saline solution) as the negative control for the treatment. After 24 hours, the cells were lysed, and their RNA was extracted for RNAseq.
Project description:HEK-293T cells were plated (1x106 cells) and treated with a nutraceutical formula (solution-3) at 0.01x concentration or with vehicle (saline solution) as the negative control for the treatment. After 24 hours, the cells were lysed, and their RNA was extracted for RNAseq.
Project description:Primary human nasal epithelial cells collected by nasal brushing of healthy donors (data deposited at: EVA- EMBL-EBI; project ID: PRJEB42411; analyses: ERZ1700617) were plated (3x105 cells) and infected with SARS-CoV-2 viral particles 20A (EPI_ISL_514432-S66; MOI, 0.03). After 24 hours, the infected cells were treated with 18.5 microMolar long chain inorganic polyphosphates (polyP120) or vehicle (saline solution) as the negative control for the treatment. After 36 hours, the cells were lysed, and their RNA were extracted for RNAseq.
Project description:Physical interactions between viral and host proteins are responsible for almost all aspects of the viral life cycle and the host’s immune response. Studying viral-host protein-protein interactions is thus crucial for identifying strategies for treatment and prevention of viral infection. Here, we use high-throughput yeast two-hybrid and affinity purification followed by mass spectrometry to generate a comprehensive SARS-CoV-2-human protein-protein interactome network consisting of both binary and co-complex interactions. We report a total of 739 high-confidence interactions, showing the highest overlap of interaction partners among published datasets as well as the highest overlap with genes differentially expressed in samples (such as upper airway and bronchial epithelial cells) from patients with SARS-CoV-2 infection. Showcasing the utility of our network, we describe a novel interaction between the viral accessory protein ORF3a and the host zinc finger transcription factor ZNF579 to illustrate a SARS-CoV-2 factor mediating a direct impact on host transcription. Leveraging our interactome, we performed network-based drug screens for over 2,900 FDA-approved/investigational drugs and obtained a curated list of 23 drugs that had significant network proximities to SARS-CoV-2 host factors, one of which, carvedilol, showed promising antiviral properties. We performed electronic health record-based validation using two independent large-scale, longitudinal COVID-19 patient databases and found that carvedilol usage was associated with a significantly lowered probability (17%-20%, P < 0.001) of obtaining a SARS-CoV-2 positive test after adjusting various confounding factors. Carvedilol additionally showed anti-viral activity against SARS-CoV-2 in a human lung epithelial cell line [(half maximal effective concentration (EC 50 ) value of 4.1 µM]), suggesting a mechanism for its beneficial effect in COVID-19. Our study demonstrates the value of large-scale network systems biology approaches for extracting biological insight from complex biological processes.
Project description:Microcystins (MCs) are cyclic hepatotoxins produced worldwide by various species of cyanobacteria. Their structure includes two variable amino acids (AAs) and most of the studies focused on the most toxic variant: the microcystin LR (MC-LR). However, more than 80 MC variants have been described to date. Despite ingestion being the major pathway of human exposure, few in vivo studies have demonstrated macroscopic effects on the gastro-intestinal tract, but no data are available on the affected pathways by several variants on intestinal cells. Here, using a non-selective method, we investigated for the first time the effect of MC-RR and MC-LR on the human intestinal cell line Caco-2 and compared their response at the pangenomic scale. The cells were incubated for 4 hrs or 24 hrs with the same range of sub-lethal concentrations of MC-RR or MC-LR. Low effects were observed for both variants after a short-term exposure. On the contrary, dose-dependent modulations of the genes transcription levels were noticed with MC-RR and MC-LR after 24 hrs. Furthermore, the genomic profiles induced by both variants were similar suggesting a common toxicity mechanism but with higher modulation following MC-LR than MC-RR exposure. However, the functional annotation revealed major differences between the variants effects. Indeed, the well-known MC-LR affected mainly two pathways, the oxidative stress response and the cell cycle regulation, which did not elicit significant alteration following MC-RR exposure. This work is the first comparative description of the MC-LR and MC-RR effects on a human intestinal cell model. It allowed us to suggest differences in the mechanism of toxicity for MC-RR and MC-LR. These results illustrate that the toxicity of MC variants remains a key point for risk assessment. Differentiated Caco-2 cells were exposed to microcystins in free FCS culture medium for either 4 or 24 hours. Sub-lethal concentrations of 10, 50 and 100 M-BM-5M of MC-LR or MC-RR were chosen for 4 hours, while 1, 5 and 10 M-BM-5M were selected for 24 hours. For each condition (including the controls), the solvent concentration was fixed to 2% EtOH for MC-LR and 1.5% of 80% MeOH for MC-RR. Four to five culture replicates per condition were done.
Project description:We discovered that cellular uptake of sorbate, an FDA-approved and widely used food preservative, can induce lysine sorbylation (Ksor), a new posttranslational modification and epigenetic mark. Proteomics analysis identified over 20 histone sorbylation sites in cells and mouse liver as well as over 900 proteins in Raw264.7 cells as lysine sorbylation targets.
Project description:Ceramide synthases are central enzymes of the sphingolipid pathway and are important for the production of sphingolipids of different chain length. Sphingolipids of different chain length impact the physicochemical properties of membranes. Here we investigated by complexome profiling how the expression level of membrane proteins is affected in human colon cancer cells (Caco-2 and HCT-116) that were transduced with a shRNA against CerS4 or CerS5 to downregulate both enzymes.
Project description:To identify key biological pathways that define toxicity or biocompatibility after nanoparticle exposure, three human cell types were exposed in vitro to two high aspect ratio nanoparticles for 1 hr or 24 hr and collected for global transcriptomics. Transcriptional responses were measured by global microarray analysis of cells in culture. Groups (N=3 biological replicates) of Caco-2/HT29-MTX cells exposed to 0, 10 or 100 ug/ml MWCNT or TiO2-NB nanoparticles for 1 or 24 hr.
Project description:We investigated transcriptional response of CaCo-2 cells to iron treatments, we studied hemin effect by adding hemin to DMEM-FBS medium and iron deficiency effects in using an iron free medium compared to the same supplemented with FAC (ferric ammonium citrate). Experiment Overall Design: Biological replicates were used (3 samples) of each iron treatments (SF-FAC, SF-0, DMEM-FBS, DMEM-Hemin). Each sample was labelled with cy5 and a pool of each sample was constituted and labelled with cy3.
Project description:The deposit microarray data were generated in a study that comprehensively integrated gene expression profiles and metabolic responses of Caco-2 cells that incubated with either E. coli K-12 or O157:H7. The aim of this study is to examine the impact of colonic bacteria on the global gene expression regulation and metabolite levels of the host, and investigate the molecular mechanics of the E. coli/host interaction. Caco-2 cells were incubated with one of two strains of E. coli strains (K-12 or O157:H7). Gene expression measurements were analyzed at three time points: 60, 90, and 120 minutes after co-culture, and compared with the controls in the monoculture.