MCC950/CRID3 potently targets the NACHT domain of wild-type NLRP3 but not disease-associated mutants for inflammasome inhibition

The nucleotide-binding-domain (NBD)-and leucine-rich repeat (LRR)-containing (NLR) family, pyrin-domain-containing 3 (NLRP3) inflammasome drives pathological inflammation in a suite of autoimmune, metabolic, malignant, and neurodegenerative diseases. Additionally, NLRP3 gain-of-function point mutations cause systemic periodic fever syndromes that are collectively known as cryopyrin-associated periodic syndrome (CAPS). There is significant interest in the discovery and development of diarylsulfonylurea Cytokine Release Inhibitory Drugs (CRIDs) such as MCC950/CRID3, a potent and selective inhibitor of the NLRP3 inflammasome pathway, for the treatment of CAPS and other diseases. However, drug discovery efforts have been constrained by the lack of insight into the molecular target and mechanism by which these CRIDs inhibit the NLRP3 inflammasome pathway. Here, we show that the NAIP, CIITA, HET-E, and TP1 (NACHT) domain of NLRP3 is the molecular target of diarylsulfonylurea inhibitors. Interestingly, we find photoaffinity labeling (PAL) of the NACHT domain requires an intact (d)ATP-binding pocket and is substantially reduced for most CAPS-associated NLRP3 mutants. In concordance with this finding, MCC950/CRID3 failed to inhibit NLRP3-driven inflammatory pathology in two mouse models of CAPS. Moreover, it abolished circulating levels of interleukin (IL)-1 beta and IL-18 in lipopolysaccharide (LPS)-challenged wild-type mice but not in Nlrp3(L351P) knock-in mice and ex vivo-stimulated mutant macrophages. These results identify wild-type NLRP3 as the molecular target of MCC950/CRID3 and show that CAPS-related NLRP3 mutants escape efficient MCC950/CRID3 inhibition. Collectively, this work suggests that MCC950/CRID3-based therapies may effectively treat inflammation driven by wild-type NLRP3 but not CAPS-associated mutants

Overview and Future Developments of the intelligent, FPGA-based DAQ (iFDAQ) of COMPASS

Modern experiments in high energy physics impose great demands on reliability, efficiency, and data rate of Data Acquisition Systems (DAQ). In order to address these needs, we present a versa- tile and scalable DAQ which executes the event building task entirely in FPGA modules. In 2014, the intelligent FPGA-based DAQ (iFDAQ) was deployed at the COMPASS experiment located at the Super Proton Synchrotron (SPS) at CERN. The core of the iFDAQ is its hardware Event Builder (EB), which consists of up to nine custom designed FPGA modules complying with the μ TCA/AMC standard. The EB replaced 30 distributed online computers and around 100 PCI cards increasing compactness, scalability, reliability, and bandwidth compared to the previous system. The iFDAQ in the configuration of COMPASS provides a bandwidth of up to 500 MB/s of sustained rate. By buffering data on different levels, the system exploits the spill structure of the SPS beam and averages the maximum on-spill data rate of 1.5 GB/s over the whole SPS duty cycle. It can even handle peak data rates of 8 GB/s. Its Run Control Configuration and Readout (RCCAR) software offers native user-friendly control and monitoring tools and together with the firmware of the modules provides built-in intelligence like self-diagnostics, data consis- tency checks, and front-end error handling. From 2017, all involved point-to-point high-speed links between front-end electronics, the hardware EB, and the readout computers will be wired via a passive programmable crosspoint switch. Thus, multiple event building topologies can be configured to adapt to different system sizes and communication patterns