Hepatitis C virus infection and related liver disease: the quest for the best animal model

Hepatitis C virus (HCV) is a major cause of cirrhosis and hepatocellular carcinoma (HCC) making the virus the most common cause of liver failure and transplantation. HCV is estimated to chronically affect 130 million individuals and to lead to more than 350,000 deaths per year worldwide. A vaccine is currently not available. The recently developed direct acting antivirals (DAAs) have markedly increased the efficacy of the standard of care but are not efficient enough to completely cure all chronically infected patients and their toxicity limits their use in patients with advanced liver disease, co-morbidity or transplant recipients. Because of the host restriction, which is limited to humans and non-human primates, in vivo study of HCV infection has been hampered since its discovery more than 20 years ago. The chimpanzee remains the most physiological model to study the innate and adaptive immune responses, but its use is ethically difficult and is now very restricted and regulated. The development of a small animal model that allows robust HCV infection has been achieved using chimeric liver immunodeficient mice, which are therefore not suitable for studying the adaptive immune responses. Nevertheless, these models allowed to go deeply in the comprehension of virus-host interactions and to assess different therapeutic approaches. The immunocompetent mouse models that were recently established by genetic humanization have shown an interesting improvement concerning the study of the immune responses but are still limited by the absence of the complete robust life cycle of the virus. In this review, we will focus on the relevant available animal models of HCV infection and their usefulness for deciphering the HCV life cycle and virus-induced liver disease, as well as for the development and evaluation of new therapeutics. We will also discuss the perspectives on future immunocompetent mouse models and the hurdles to their development

Three-dimensional instability of a ow past a sphere: Mach evolution of the regular and Hopf bifurcations

A fully three-dimensional linear stability analysis is carried out to investigate the unstable bifurcations of a compressible viscous fluid past a sphere. A time-stepper technique is used to compute both equilibrium states and leading eigenmodes. In agreement with previous studies, the numerical results reveal a regular bifurcation under the action of a steady mode and a supercritical Hopf bifurcation that causes the onset of unsteadiness but also illustrate the limitations of previous linear approaches, based on parallel and axisymmetric base flow assumptions, or weakly nonlinear theories. The evolution of the unstable bifurcations is investigated up to low-supersonic speeds. For increasing Mach numbers, the thresholds move towards higher Reynolds numbers. The unsteady fluctuations are weakened and an axisymmetrization of the base flow occurs. For a sufficiently high Reynolds number, the regular bifurcation disappears and the flow directly passes from an unsteady planar-symmetric solution to a stationary axisymmetric stable one when the Mach number is increased. A stability map is drawn by tracking the bifurcation boundaries for different Reynolds and Mach numbers. When supersonic conditions are reached, the flow becomes globally stable and switches to a noise-amplifier system. A continuous Gaussian white noise forcing is applied in front of the shock to examine the convective nature of the flow. A Fourier analysis and a dynamic mode decomposition show a modal response that recalls that of the incompressible unsteady cases. Although transition in the wake does not occur for the chosen Reynolds number and forcing amplitude, this suggests a link between subsonic and supersonic dynamics

Numerical investigation of sheet cavitation over a 3-D venturi configuration

Sheet cavitation appears in many hydraulic applications and can lead to technical issues. Numerical simulation is a pertinent way to study the phenomenon. A numerical tool based on 1-fluid compressible RANS equations with a cavitation model is used to compute a flow within a 3-D venturi geometry with a 4° divergent angle. In the present work, a detailed study of this cavitating flow, which presents a quasi-stable vapour pocket, is carried out using tools such as Power Spectral Densities or Spectral Proper Orthogonal Decompositions. An oblique oscillation of the cavity is then identified and discussed

Contribution à la caractérisation de sites sableux : signature spectro-directionnelle, distribution en taille et minéralogie extraites d'échantillons de sables

International audienceThe characterization of sands detailed in this paper has been performed in order to support the in-flight radiometric performance assessment of space-borne optical sensors over so-called Pseudo-Invariant Calibration Sites (PICS). Although the physical properties of PICS surface are fairly stable in time, the signal measured from space varies with the illumination and the viewing geometries. Thus there is a need to characterize the spectro-directional properties of PICS. This can be done, at a broad scale, thanks to multi-spectral multi-directional space-borne sensors such as the POLDER instrument (with old data). However, interpolating or extrapolating the spectro-directional reflectances measured from space to spectral bands of another sensor is not straightforward. The hyperspectral characterization of sand samples collected within or nearby PICS can contribute to a solution. In this context, a set of 31 sand samples was compiled. The BiConical Reflectance Factor (BCRF) was measured between 0.4 and 2.5 µm, over a quarter hemisphere when the amount of sand in the sample was large enough and for only a single fixed angular configuration for small samples. These optical measurements were complemented by grain size distribution measurements and mineralogical analysis and compiled together with previously published measurements in the so-called PICSAND database, freely available on line.La caractérisation des sables détaillée dans cet article a été faite en soutien à l'estimation en vol des performances radiométriques des capteurs optiques spatiaux à partir des sites appelés PICS pour Pseudo-Invariant Calibration Sites. Bien que les propriétés physiques des PICS soient relativement stables dans le temps, le signal mesuré depuis l'espace varie en fonction des géométries d'illumination et d'observation. De ce fait, il est nécessaire de caractériser les propriétés spectro-directionnelles des PICS. Ceci peut être fait, à une grande échelle, à partir de capteurs spatiaux multi-spectraux et multi-directionnels tels que le capteur POLDER (avec des données anciennes). Cependant, l'interpolation ou l'extrapolation des réflectances spectro-directionnelles obtenues depuis l'espace aux bandes spectrales d'un autre capteur est délicate. La caractérisation hyperspectrale d'échantillons de sable issus de PICS ou de leur voisinage peut participer à une solution. Dans ce contexte, 31 échantillons de sable ont été collectés. Le Facteur de Reflectance BiConique (BCRF) a été mesuré entre 0,4 et 2,5 µm, pour une demi-hémisphère lorsque la quantité de sable était suffisante, et pour une seule géométrie pour les échantillons plus petits. Ces mesures optiques ont été complétées par des mesures de distribution en taille et par une analyse minéralogique, et mises dans une base de données appelée PICSAND avec d'autres mesures publiées dans la littérature. Cette base de donnée est en libre accès en ligne

Nat. Hazards Earth Syst. Sci.

International audienceThe two primary causes of surf zone injuries (SZIs) worldwide, including fatal drowning and severe spinal injuries, are rip currents (rips) and shore-break waves. SZIs also result from surfing and bodyboarding activity. In this paper we address the primary environmental controls on SZIs along the high-energy meso-macro-tidal surf beach coast of southwestern France. A total of 2523 SZIs recorded by lifeguards over 186 sample days during the summers of 2007, 2009 and 2015 were combined with measured and/or hindcast weather, wave, tide, and beach morphology data. All SZIs occurred disproportionately on warm sunny days with low wind, likely because of increased beachgoer numbers and hazard exposure. Relationships were strongest for shore-break- and rip-related SZIs and weakest for surfingrelated SZIs, the latter being also unaffected by tidal stage or range. Therefore, the analysis focused on bathers. More shore-break-related SZIs occur during shore-normal incident waves with average to below-average wave height (significant wave height, Hs = 0.75-1.5 m) and around higher water levels and large tide ranges when waves break on the steepest section of the beach. In contrast, more rip-related drownings occur near neap low tide, coinciding with maximised channel rip flow activity, under shore-normal incident waves with Hs > 1.25 m and mean wave periods longer than 5 s. Addi- tional drowning incidents occurred at spring high tide, presumably due to small-scale swash rips. The composite wave and tide parameters proposed by Scott et al. (2014) are key controlling factors determining SZI occurrence, although the risk ranges are not necessarily transferable to all sites. Summer beach and surf zone morphology is interannually highly variable, which is critical to SZI patterns. The upper beach slope can vary from 0.06 to 0.18 between summers, resulting in low and high shore-break-related SZIs, respectively. Summers with coast-wide highly (weakly) developed rip channels also result in widespread (scarce) rip-related drowning incidents. With life risk defined in terms of the number of people exposed to life threatening hazards at a beach, the ability of morphodynamic models to simulate primary beach morphology characteristics a few weeks or months in advance is therefore of paramount importance for predicting the primary surf zone life risks along this coast

SHORT syndrome due to a novel de novo mutation in PRKCE (Protein Kinase Cɛ) impairing TORC2-dependent AKT activation.

SHORT syndrome is a rare, recognizable syndrome resulting from heterozygous mutations in PIK3R1 encoding a regulatory subunit of phosphoinositide-3-kinase (PI3K). The condition is characterized by short stature, intrauterine growth restriction, lipoatrophy and a facial gestalt involving a triangular face, deep set eyes, low hanging columella and small chin. PIK3R1 mutations in SHORT syndrome result in reduced signaling through the PI3K-AKT-mTOR pathway. We performed whole exome sequencing for an individual with clinical features of SHORT syndrome but negative for PIK3R1 mutation and her parents. A rare de novo variant in PRKCE was identified. The gene encodes PKCε and, as such, the AKT-mTOR pathway function was assessed using phospho-specific antibodies with patient lymphoblasts and following ectopic expression of the mutant in HEK293 cells. Kinase analysis showed that the variant resulted in a partial loss-of-function. Whilst interaction with PDK1 and the mTORC2 complex component SIN1 was preserved in the mutant PKCε, it bound to SIN1 with a higher affinity than wild-type PKCε and the dynamics of mTORC2-dependent priming of mutant PKCε was altered. Further, mutant PKCε caused impaired mTORC2-dependent pAKT-S473 following rapamycin treatment. Reduced pFOXO1-S256 and pS6-S240/244 levels were also observed in the patient LCLs. To date, mutations in PIK3R1 causing impaired PI3K-dependent AKT activation are the only known cause of SHORT syndrome. We identify a SHORT syndrome child with a novel partial loss-of-function defect in PKCε. This variant causes impaired AKT activation via compromised mTORC2 complex function

Synergy of entry inhibitors with direct-acting antivirals uncovers novel combinations for prevention and treatment of hepatitis C

Objective: Although direct-acting antiviral agents (DAAs) have markedly improved the outcome of treatment in chronic HCV infection, there continues to be an unmet medical need for improved therapies in difficult-to-treat patients as well as liver graft infection. Viral entry is a promising target for antiviral therapy. Design: Aiming to explore the role of entry inhibitors for future clinical development, we investigated the antiviral efficacy and toxicity of entry inhibitors in combination with DAAs or other host-targeting agents (HTAs). Screening a large series of combinations of entry inhibitors with DAAs or other HTAs, we uncovered novel combinations of antivirals for prevention and treatment of HCV infection. Results: Combinations of DAAs or HTAs and entry inhibitors including CD81-, scavenger receptor class B type I (SR-BI)- or claudin-1 (CLDN1)-specific antibodies or small-molecule inhibitors erlotinib and dasatinib were characterised by a marked and synergistic inhibition of HCV infection over a broad range of concentrations with undetectable toxicity in experimental designs for prevention and treatment both in cell culture models and in human liver-chimeric uPA/SCID mice. Conclusions: Our results provide a rationale for the development of antiviral strategies combining entry inhibitors with DAAs or HTAs by taking advantage of synergy. The uncovered combinations provide perspectives for efficient strategies to prevent liver graft infection and novel interferon-free regimens

Performance of Pyridylthiourea-Polyethylenimine Polyplex for siRNA-Mediated Liver Cancer Therapy in Cell Monolayer, Spheroid, and Tumor Xenograft Models

Medical application of siRNAs relies on methods for delivering nucleic acids into the cytosol. Synthetic carriers, which assemble with nucleic acids into delivery systems, show promises for cancer therapy but efficiency remains to be improved. In here, the effectiveness of pyridylthiourea‐polyethylenimine (πPEI), a siRNA carrier that favors both polyplex disassembly and endosome rupture upon sensing the acidic endosomal environment, in 3 experimental models of hepatocellular cancer is tested. The πPEI‐assisted delivery of a siRNA targeting the polo‐like kinase 1 into Huh‐7 monolayer produces a 90% cell death via a demonstrated RNA interference mechanism. Incubation of polyplex with Huh‐7 spheroids leads to siRNA delivery into the superficial first cell layer and a 60% reduction in spheroid growth compared to untreated controls. Administration of polyplexes into mice bearing subcutaneous implanted Huh‐7Luc tumors results in a reduced tumor progression, similar to the one observed in the spheroid model. Altogether, these results support the in vivo use of synthetic and dedicated polymers for increasing siRNA‐mediated gene knockdown, and their clinical promise in cancer therapeutics

Non-invasive quantitative imaging of hepatocellular carcinoma growth in mice by micro-CT using liver-targeted iodinated nano-emulsions

Hepatocellular carcinoma (HCC) is the only cancer for which non-invasive diagnosis is recognized by international guidelines. Contrast agent free ultrasound imaging, computed tomography (CT) and/or magnetic resonance imaging are techniques used for early detection and confirmation. Clinical evidence depicts that CT is 30% less precise as compared to MRI for detection of small tumors. In our work, we have reported some novel tools that can enhance the sensitivity and precision of CT applied to preclinical research (micro-CT). Our system, containing non-toxic nano-droplets loaded with iodine has high contrasting properties, liver and hepatocyte specificity and strong liver persistence. Micro-CT was performed on HCC model implanted in nude mice by intrahepatic injection. Contrast agent was administrated intravenously. This method allows an unprecedented high precision of detection, quantitative measurement of tumor volume and quantitative follow-up of the tumor development.PMC565532

De novo TBR1 variants cause a neurocognitive phenotype with ID and autistic traits:report of 25 new individuals and review of the literature

TBR1, a T-box transcription factor expressed in the cerebral cortex, regulates the expression of several candidate genes for autism spectrum disorders (ASD). Although TBR1 has been reported as a high-confidence risk gene for ASD and intellectual disability (ID) in functional and clinical reports since 2011, TBR1 has only recently been recorded as a human disease gene in the OMIM database. Currently, the neurodevelopmental disorders and structural brain anomalies associated with TBR1 variants are not well characterized. Through international data sharing, we collected data from 25 unreported individuals and compared them with data from the literature. We evaluated structural brain anomalies in seven individuals by analysis of MRI images, and compared these with anomalies observed in TBR1 mutant mice. The phenotype included ID in all individuals, associated to autistic traits in 76% of them. No recognizable facial phenotype could be identified. MRI analysis revealed a reduction of the anterior commissure and suggested new features including dysplastic hippocampus and subtle neocortical dysgenesis. This report supports the role of TBR1 in ID associated with autistic traits and suggests new structural brain malformations in humans. We hope this work will help geneticists to interpret TBR1 variants and diagnose ASD probands