Project description:Reveal differentially regulated genes and cellular pathways within allergic and non-allergic asthmatic children compared to healthy controls Peripheral blood mononuclear cells (PBMCs) were obtained from allergic asthmatics (n=14), non-allergic asthmatics (n=8) and healthy controls (n=14) and kept with anti-CD3/CD28 (CD328), LpA or without stimulation (M).
Project description:VAF347 is a low molecular weight compound which inhibits allergic lung inflammation in vivo. This effect is likely due to a block of dendritic cell (DC) function to generate pro-inflammatory T-helper (Th) cells since VAF347 inhibits IL-6, CD86 and HLA-DR expression by human monocyte derived DC, three relevant molecules for Th-cell generation. Here we demonstrate that VAF347 interacts with the aryl hydrocarbon receptor (AhR) protein resulting in activation of the AhR signaling pathway. Functional AhR is responsible for the biological activity of VAF347 since, i) other AhR agonists display an identical activity profile in vitro, ii) gene silencing of wild type AhR expression or forced over-expression of a trans-dominant negative AhR ablates VAF347 activity to inhibit cytokine induced IL-6 expression in a human monocytic cell line and iii) AhR deficient mice are resistant to the compoundâs ability to block allergic lung inflammation in vivo. These data identify the AhR protein as key molecular target of VAF347 and its essential role for mediating the anti-inflammatory effects of the compound in vitro and in vivo. Experiment Overall Design: Immature monocyte-derived DC were activated with anti-CD40 antibodies for 8 hours in the absence or presence of VAF347. Two donors (D1,D2) were used, resulting in four data sets: D1_ctrl, D2_ctrl (ctrl=no treatment), D1_VAF347, D2_VAF347.
Project description:This study aims to elucidate the molecular mechanisms underlying the therapeutic effects of mountain-grown ginseng in allergic asthma and to systematically identify and characterize bioactive monomeric compounds responsible for these effects. We found that total saponins from mountain-grown ginseng significantly alleviated asthmatic phenotypes. Further multi-omics analyses revealed that these effects may be mediated through modulation of the arachidonic acid metabolic pathway, particularly by regulating the enzymatic activity of the rate-limiting enzyme PTGS2, thereby suppressing the production of PGE2. Subsequent component identification using high-performance liquid chromatography, combined with computational modeling, identified ginsenoside Rd2 as a key bioactive compound. In vitro cellular experiments, together with site-directed mutagenesis of the predicted binding sites, further confirmed that Rd2 exerts inhibitory effects on PTGS2 enzymatic activity through specific molecular interactions. Collectively, these findings provide mechanistic insights into the anti-asthmatic effects of mountain-grown ginseng and highlight Rd2 as a promising bioactive compound targeting PTGS2, offering potential therapeutic value for allergic asthma.
Project description:SARS-CoV-2 virus mimics host mRNA by capping its viral RNA to promote replication and evade host immune sensing. SARS-CoV-2 NSP14 is the N7-guanosine methyltransferase (N7-MTase) responsible for RNA cap-0 formation. Targeting NSP14 for antiviral drug development is an under-explored but promising strategy. Here we conducted a high-throughput screening on natural products library derived from Chinese herbal medicine to discover Emodin as a SARS-CoV-2 NSP14 inhibitor. Exploring Emodin derivatives, Questin was identified with potent cellular inhibitory activity (EC50=249 nM) against SARS-CoV-2, which inhibits NSP14 in an RNA cap competitive manner, making it one the most potent anti-coronaviral natural products. Mechanistically, besides catalyzing viral RNA capping, NSP14 by itself could remodel host transcriptome such as enriching CREBBP, a key host factor in cellular cyclic AMP response pathway, to promote viral infection. As a result, targeting NSP14 by Questin significantly impairs viral Replication & Translation step and reverses host transcriptome remodeled by NSP14. We next validated Questin as a promising lead with significantly improved toxicity upon acute exposure in zebrafish larvae. Taken together, our study not only demonstrates Questin as a potent drug lead for clinical antiviral application, but also highlights multiple antiviral potentials of NSP14 as therapeutic target.