Differential IL-1β secretion by monocyte subsets is regulated by Hsp27 through modulating mRNA stability.

Monocytes play a central role in regulating inflammation in response to infection or injury, and during auto-inflammatory diseases. Human blood contains classical, intermediate and non-classical monocyte subsets that each express characteristic patterns of cell surface CD16 and CD14; each subset also has specific functional properties, but the mechanisms underlying many of their distinctive features are undefined. Of particular interest is how monocyte subsets regulate secretion of the apical pro-inflammatory cytokine IL-1β, which is central to the initiation of immune responses but is also implicated in the pathology of various auto-immune/auto-inflammatory conditions. Here we show that primary human non-classical monocytes, exposed to LPS or LPS + BzATP (3'-O-(4-benzoyl)benzyl-ATP, a P2X7R agonist), produce approx. 80% less IL-1β than intermediate or classical monocytes. Despite their low CD14 expression, LPS-sensing, caspase-1 activation and P2X7R activity were comparable in non-classical monocytes to other subsets: their diminished ability to produce IL-1β instead arose from 50% increased IL-1β mRNA decay rates, mediated by Hsp27. These findings identify the Hsp27 pathway as a novel therapeutic target for the management of conditions featuring dysregulated IL-1β production, and represent an advancement in understanding of both physiological inflammatory responses and the pathogenesis of inflammatory diseases involving monocyte-derived IL-1β

RNA-based gene therapy for haemophilia B

Haemophilia B, a deficiency in clotting factor IX (FIX), occurs in about I in 25.000 males. Of these patients, approximately 40% are characterized as having 'severe' haemophilia (FIX below 1% of normal). Although the use of plasma-derived or rFIX has extended the lifespan of these patients, they remain afflicted by a variety of sequelae of the disease. Interestingly, a relatively small increase in the levels of FIX results in a dramatic increase in the quality of life, requiring rFIX substitution only prior to dental and surgical procedures. This latter trait of the disease suggests that gene therapy only needs to achieve modest expression and makes this disease a prime candidate for proof-of-concept of gene therapy. Current protocols for gene therapy entail the risk of malignant transformation. In addition the viral vectors may cause substantial pathology. Hence, the field requires the development of inherently safe gene therapy. Recently, developed protocols have been developed to transduce peripheral blood cells with high efficiency using RNA as a vector. These protocols in which the inherently safe RNA-based gene therapy approach will be validated and implemented for haemophilia B will provide in turn providing important proof-of concept for this type of therapy for more prevalent diseases but requires optimization of this strategy, probably by enhancing RNA stability using artificial nucleotides and modified UTRs, enhanced production of protein by codon usage optimization and the use of multi-cistronic constructs, and enhanced secretion of the protein employing target cell-tailored secretion signals.</p

RNA-based gene therapy for haemophilia B

Haemophilia B, a deficiency in clotting factor IX (FIX), occurs in about I in 25.000 males. Of these patients, approximately 40% are characterized as having 'severe' haemophilia (FIX below 1% of normal). Although the use of plasma-derived or rFIX has extended the lifespan of these patients, they remain afflicted by a variety of sequelae of the disease. Interestingly, a relatively small increase in the levels of FIX results in a dramatic increase in the quality of life, requiring rFIX substitution only prior to dental and surgical procedures. This latter trait of the disease suggests that gene therapy only needs to achieve modest expression and makes this disease a prime candidate for proof-of-concept of gene therapy. Current protocols for gene therapy entail the risk of malignant transformation. In addition the viral vectors may cause substantial pathology. Hence, the field requires the development of inherently safe gene therapy. Recently, developed protocols have been developed to transduce peripheral blood cells with high efficiency using RNA as a vector. These protocols in which the inherently safe RNA-based gene therapy approach will be validated and implemented for haemophilia B will provide in turn providing important proof-of concept for this type of therapy for more prevalent diseases but requires optimization of this strategy, probably by enhancing RNA stability using artificial nucleotides and modified UTRs, enhanced production of protein by codon usage optimization and the use of multi-cistronic constructs, and enhanced secretion of the protein employing target cell-tailored secretion signals

RNA-based gene therapy for haemophilia B

Haemophilia B, a deficiency in clotting factor IX (FIX), occurs in about I in 25.000 males. Of these patients, approximately 40% are characterized as having 'severe' haemophilia (FIX below 1% of normal). Although the use of plasma-derived or rFIX has extended the lifespan of these patients, they remain afflicted by a variety of sequelae of the disease. Interestingly, a relatively small increase in the levels of FIX results in a dramatic increase in the quality of life, requiring rFIX substitution only prior to dental and surgical procedures. This latter trait of the disease suggests that gene therapy only needs to achieve modest expression and makes this disease a prime candidate for proof-of-concept of gene therapy. Current protocols for gene therapy entail the risk of malignant transformation. In addition the viral vectors may cause substantial pathology. Hence, the field requires the development of inherently safe gene therapy. Recently, developed protocols have been developed to transduce peripheral blood cells with high efficiency using RNA as a vector. These protocols in which the inherently safe RNA-based gene therapy approach will be validated and implemented for haemophilia B will provide in turn providing important proof-of concept for this type of therapy for more prevalent diseases but requires optimization of this strategy, probably by enhancing RNA stability using artificial nucleotides and modified UTRs, enhanced production of protein by codon usage optimization and the use of multi-cistronic constructs, and enhanced secretion of the protein employing target cell-tailored secretion signals.6640140

Synchrotron-microtomographic studies of normal and pathological cranial sutures - a further insight

ADAM17 is implicated in several debilitating diseases. However, drug discovery efforts targeting ADAM17 have failed due to the utilization of zinc-binding inhibitors. We previously reported discovery of highly selective nonzinc-binding exosite-targeting inhibitors of ADAM17 that exhibited not only enzyme isoform selectivity but synthetic substrate selectivity as well ( J. Biol. Chem. 2013, 288, 22871). As a result of SAR studies presented herein, we obtained several highly selective ADAM17 inhibitors, six of which were further characterized in biochemical and cell-based assays. Lead compounds exhibited low cellular toxicity and high potency and selectivity for ADAM17. In addition, several of the leads inhibited ADAM17 in a substrate-selective manner, which has not been previously documented for inhibitors of the ADAM family. These findings suggest that targeting exosites of ADAM17 can be used to obtain highly desirable substrate-selective inhibitors. Additionally, current inhibitors can be used as probes of biological activity of ADAM17 in various in vitro and, potentially, in vivo systems