Drosophila H2Av negatively regulates the activity of the IMD pathway via facilitating Relish SUMOylation

Insects depend on the innate immune response for defense against a wide array of pathogens. Central to Drosophila immunity are antimicrobial peptides (AMPs), released into circulation when pathogens trigger either of the two widely studied signal pathways, Toll or IMD. The Toll pathway responds to infection by Gram-positive bacteria and fungi while the IMD pathway is activated by Gram-negative bacteria. During activation of the IMD pathway, the NF-κB-like transcription factor Relish is phosphorylated and then cleaved, which is crucial for IMD-dependent AMP gene induction. Here we show that loss-of-function mutants of the unconventional histone variant H2Av upregulate IMD-dependent AMP gene induction in germ-free Drosophila larvae and adults. After careful dissection of the IMD pathway, we found that Relish has an epistatic relationship with H2Av. In the H2Av mutant larvae, SUMOylation is down-regulated, suggesting a possible role of SUMOylation in the immune phenotype. Eventually we demonstrated that Relish is mostly SUMOylated on amino acid K823. Loss of the potential SUMOylation site leads to significant auto-activation of Relish in vivo. Further work indicated that H2Av regulates Relish SUMOylation after physically interacting with Su(var)2-10, the E3 component of the SUMOylation pathway. Biochemical analysis suggested that SUMOylation of Relish prevents its cleavage and activation. Our findings suggest a new mechanism by which H2Av can negatively regulate, and thus prevent spontaneous activation of IMD-dependent AMP production, through facilitating SUMOylation of the NF-κB like transcription factor Relish.

MO097BETA BLOCKER PREVENTS CARDIAC MOLECULAR AND MORPHOLOGICAL REMODELLING IN EXPERIMENTAL URAEMIA

Background and Aims Fifty years of heart failure research has shown that pathological cardiac remodelling forms a vicious cycle with myocardial dysfunction leading to progressive heart failure (HF) [Circulation, 102 IV14-23, 2000]. Fetal gene induction is associated with this process and beta blocker therapy has been shown to prevent it. Although chronic kidney disease (CKD) and HF share similar mediators of cardiac remodelling, the benefits of beta blocker therapy in CKD has not been studied. We, therefore, tested the hypothesis that beta blocker therapy prevents fetal gene induction and pathological cardiac remodelling in experimental uraemia. Method Wistar rats (n=32) had subtotal nephrectomy (STNx) [Frontiers in physiology, 10 1365, 2019] or sham surgery and were followed up for 10 weeks. The animals were randomly allocated to metoprolol (10mg/kg/day) or vehicle. In vivo and in vitro cardiac assessments were performed, and changes in myocardial fetal gene expression were also studied. Results Heart rate was significantly lower in metoprolol groups compared to untreated groups demonstrating effective beta blockade (Fig 1A). Echocardiographic LV mass was significantly higher in untreated STNx group compared to the metoprolol group (896.4 vs 632.2g, P=0.0004). Similar changes were seen with heart weight to tibia ratio (Fig 1B). There was no significant difference in blood pressure (BP) between treated and untreated STNx animals (123 vs 119 mmHg, P=0.359) (Fig 1A). STNx increased mRNA expression of fetal genes and there was a trend towards attenuation of this increase with beta blocker therapy (Fig 1C). Conclusion Beta blocker therapy ameliorates uraemic pathological cardiac remodelling irrespective of changes to BP. This benefit appears be associated with a reduction of induced fetal gene expression. Further translational research on the benefits of beta blockade in the treatment of uraemic cardiomyopathy is required.

Egr2 induction in SPNs of the ventrolateral striatum contributes to cocaine place preference in mice

Drug addiction develops due to brain-wide plasticity within neuronal ensembles, mediated by dynamic gene expression. Though the most common approach to identify such ensembles relies on immediate early gene expression, little is known of how the activity of these genes is linked to modified behavior observed following repeated drug exposure. To address this gap, we present a broad-to-specific approach, beginning with a comprehensive investigation of brain-wide cocaine-driven gene expression, through the description of dynamic spatial patterns of gene induction in subregions of the striatum, and finally address functionality of region-specific gene induction in the development of cocaine preference. Our findings reveal differential cell-type specific dynamic transcriptional recruitment patterns within two subdomains of the dorsal striatum following repeated cocaine exposure. Furthermore, we demonstrate that induction of the IEG Egr2 in the ventrolateral striatum, as well as the cells within which it is expressed, are required for the development of cocaine seeking.