pinecone/core-2020-05-10-deduplication huggingface dataset:
A phase 2, randomized, double‐blind, placebo‐controlled study of GS‐9450 in subjects with nonalcoholic steatohepatitis

In nonalcoholic steatohepatitis (NASH), the extent of hepatocyte apoptosis correlates with disease severity. Reducing hepatocyte apoptosis with the selective caspase inhibitor GS‐9450 has a potential for altering the course of the liver disease. In this phase 2, double‐blind study, 124 subjects with biopsy‐proven NASH were randomized to once‐daily placebo or 1, 5, 10, or 40 mg GS‐9450 for 4 weeks. Absolute and percent changes from baseline in ALT levels, AST levels, and caspase‐3–cleaved cytokeratin (CK)‐18 fragments at week 4 were assessed by an analysis of covariance model with adjustment for baseline values. In the 40‐mg group, mean (SD) ALT decreased by 47 (43) U/L from baseline to week 4 ( P  < 0.0001 versus placebo), and the proportion of subjects with normal ALT increased from 0% to 35% at week 4. In the 40‐mg group, mean AST decreased by 13 U/L from baseline (not significant), and the proportion with normal AST increased from 20% at baseline to 48% at week 4. By week 4, mean CK‐18 fragment levels had decreased to 393 (723) U/L in the GS‐9450 10‐mg group and 125 (212) U/L in the 40‐mg group, but these reductions were not statistically significant. No serious adverse events were reported during treatment, and the percentage of subjects with at least one treatment‐emergent grade 3 or 4 laboratory abnormality ranged from 11.5% to 17% across the GS‐9450 treatment groups versus 35% in the placebo group.  Conclusion : GS‐9450 treatment induced significant reductions in ALT levels in NASH patients. Reductions in CK‐18 fragment levels also occurred, although they were not statistically significant. At appropriate therapeutic indices, selective caspase inhibitors may be a promising treatment option in patients with NASH. (H epatology  2012
Glucokinase links Krüppel-like factor 6 to the regulation of hepatic insulin sensitivity in nonalcoholic fatty liver disease.

The polymorphism, KLF6-IVS1-27A, in the Krüppel-like factor 6 (KLF6) transcription factor gene enhances its splicing into antagonistic isoforms and is associated with delayed histological progression of nonalcoholic fatty liver disease (NAFLD). To explore a potential role for KLF6 in the development of insulin resistance, central to NAFLD pathogenesis, we genotyped KLF6-IVS1-27 in healthy subjects and assayed fasting plasma glucose (FPG) and insulin sensitivities. Furthermore, we quantified messenger RNA (mRNA) expression of KLF6 and glucokinase (GCK), as an important mediator of insulin sensitivity, in human livers and in liver tissues derived from a murine Klf6 knockdown model (DeltaKlf6). Klf6 overexpression studies in a mouse hepatocyte line were utilized to mechanistically link KLF6 with Gck promoter activity. KLF6-IVS1-27Gwt (i.e., less KLF6 splicing) was associated with stepwise increases in FPG and insulin and reduced hepatic insulin sensitivity. KLF6 binds to the liver-specific Gck promoter and activates a GCK promoter-reporter, identifying GCK as a KLF6 direct transcriptional target. Accordingly, in DeltaKlf6 hepatocytes Gck expression was reduced and stable transfection of Klf6 led to up-regulation of Gck. GCK and KLF6 mRNAs correlate directly in human NAFLD tissues and immunohistochemistry studies confirm falling levels of both KLF6 and GCK in fat-laden hepatocytes. In contrast to full-length KLF6, splice variant KLF6-SV1 increases in NAFLD hepatocytes and inversely correlates with glucokinase regulatory protein, which negatively regulates GCK activity. Conclusion: KLF6 regulation of GCK contributes to the development of hepatic insulin resistance. The KLF6-IVS1-27A polymorphism, which generates more KLF6-SV1, combats this, lowering hepatic insulin resistance and blood glucose
Report on a single‐topic conference on “Chronic Viral Hepatitis–Strategies to Improve Effectiveness of Screening and Treatment”

The 2010 Institute of Medicine report on “Hepatitis and Liver Cancer” indicated that lack of knowledge and awareness about chronic hepatitis B (HBV) and C virus (HCV) infections and insufficient understanding about the extent and seriousness of this public health problem impeded current efforts to prevent and control hepatitis B and C. A single‐topic conference was held in June 2011 to discuss strategies to improve the effectiveness of screening, care referral, and clinical management of chronic HBV and HCV infections with the ultimate goal of reducing morbidity and mortality from these infections. Various models that have been shown to improve hepatitis screening and effectiveness of hepatitis treatment in the community, including rural settings and populations that have traditionally been excluded due to comorbidities, were presented. Recent advances in laboratory testing, medical management, and new antiviral therapies will not decrease the burden of viral hepatitis if persons at risk for or who are living with viral hepatitis are not aware of the risks, have not been diagnosed, or have no access to care. Systematic changes in our health care delivery system and enhanced coordination of prevention and care services with partnerships between public health leaders and clinicians through education of the public and health care providers and linkage of infected persons with care and treatment services can increase the success of preventing viral hepatitis and the effectiveness of hepatitis treatment in the real world. Implementation of these changes is feasible and will require policy changes, coordination among government agencies, and collaboration between government agencies, health care providers, community organizations, and advocacy groups. (H EPATOLOGY  2012;55:307–315
Peroxisome proliferator‐activated receptor alpha induction of uncoupling protein 2 protects against acetaminophen‐induced liver toxicity

Acetaminophen (APAP) overdose causes acute liver failure in humans and rodents due in part to the destruction of mitochondria as a result of increased oxidative stress followed by hepatocellular necrosis. Activation of the peroxisome proliferator‐activated receptor alpha (PPARα), a member of the nuclear receptor superfamily that controls the expression of genes encoding peroxisomal and mitochondrial fatty acid β‐oxidation enzymes, with the experimental ligand Wy‐14,643 or the clinically used fibrate drug fenofibrate, fully protects mice from APAP‐induced hepatotoxicity. PPARα‐humanized mice were also protected, whereas  Ppara ‐null mice were not, thus indicating that the protection extends to human PPARα and is PPARα‐dependent. This protection is due in part to induction of the PPARα target gene encoding mitochondrial uncoupling protein 2 (UCP2). Forced overexpression of UCP2 protected wildtype mice against APAP‐induced hepatotoxicity in the absence of PPARα activation.  Ucp2 ‐null mice, however, were sensitive to APAP‐induced hepatotoxicity despite activation of PPARα with Wy‐14,643. Protection against hepatotoxicity by UCP2‐induction through activation of PPARα is associated with decreased APAP‐induced c‐jun and c‐fos expression, decreased phosphorylation of JNK and c‐jun, lower mitochondrial H 2 O 2  levels, increased mitochondrial glutathione in liver, and decreased levels of circulating fatty acyl‐carnitines. These studies indicate that the PPARα target gene UCP2 protects against elevated reactive oxygen species generated during drug‐induced hepatotoxicity and suggest that induction of UCP2 may also be a general mechanism for protection of mitochondria during fatty acid β‐oxidation. (H EPATOLOGY  2012;56:281–290
Oxidative stress, Nrf2 and keratin up‐regulation associate with Mallory‐Denk body formation in mouse erythropoietic protoporphyria

Mallory‐Denk bodies (MDBs) are hepatocyte inclusions commonly seen in steatohepatitis. They are induced in mice by feeding 3,5‐diethoxycarbonyl‐1,4‐dihydrocollidine (DDC) for 12 weeks, which also causes porphyrin accumulation. Erythropoietic protoporphyria (EPP) is caused by mutations in ferrochelatase (fch), and a fraction of EPP patients develop liver disease that is phenocopied in Fech m1Pas  mutant (fch/fch) mice, which have an inactivating fch mutation. fch/fch mice develop spontaneous MDBs, but the molecular factors involved in their formation and whether they relate to DDC‐induced MDBs are unknown. We tested the hypothesis that fch mutation creates a molecular milieu that mimics experimental drug‐induced MDBs. In 13‐ and 20‐week‐old fch/fch mice, serum alkaline phosphatase, alanine aminotransferase, and bile acids were increased. The 13‐week‐old fch/fch mice did not develop histologically evident MDBs but manifested biochemical alterations required for MDB formation, including increased transglutaminase‐2 and keratin overexpression, with a greater keratin 8 (K8)‐to‐keratin 18 (K18) ratio, which are critical for drug‐induced MDB formation. In 20‐week‐old fch/fch mice, spontaneous MDBs were readily detected histologically and biochemically. Short‐term (3‐week) DDC feeding markedly induced MDB formation in 20‐week‐old fch/fch mice. Under basal conditions, old fch/fch mice had significant alterations in mitochondrial oxidative‐stress markers, including increased protein oxidation, decreased proteasomal activity, reduced adenosine triphosphate content, and Nrf2 (redox sensitive transcription factor) up‐regulation. Nrf2 knockdown in HepG2 cells down‐regulated K8, but not K18.  Conclusion : Fch/fch mice develop age‐associated spontaneous MDBs, with a marked propensity for rapid MDB formation upon exposure to DDC, and therefore provide a genetic model for MDB formation. Inclusion formation in the fch/fch mice involves oxidative stress which, together with Nrf2‐mediated increase in K8, promotes MDB formation. (H epatology  2012;56:322–331
Studies on the effects of thyroid hormone on cholesterol and lipoprotein metabolism

Elevated plasma lipids, particularly cholesterol within low density lipoproteins, is an important 
risk factor for developing atherosclerosis  which can cause angina pectoris, myocardial 
infarction, and stroke. Thyroid hormone (TH) has a strong influence on lipid metabolism and 
the aim of this thesis was to gain more insight into how  TH modulates cholesterol and 
lipoprotein metabolism.



  
In contrast to normal rats, plasma cholesterol increases in hypophysectomized (Hx) rats upon 
cholesterol feeding. In paper I, it was found that the increased plasma cholesterol in Hx rats in 
response to cholesterol feeding is partly caused by an increased intestinal absorption of dietary 
cholesterol. TH was found to normalize the increased absorption of cholesterol in Hx rats. The 
changes in intestinal absorption of dietary  cholesterol induced by hypophysectomy and TH 
could not be explained by changes in the intestinal gene expressions of sterol transporters 
ABCG5/G8 and NPC1L1. However, hepatic gene expressions of ABCG5/G8 were found to be 
diminished in Hx rats and strongly stimulated by TH, associated with a markedly reduced and 
stimulated biliary secretion of cholesterol, respectively, that may influence the absorption of 
dietary cholesterol.



 
The sterol transporter ABCG5/G8 is of major importance in the biliary secretion of cholesterol. 
However, alternative pathways that promote biliary cholesterol secretion have been proposed. 
TH stimulates both biliary cholesterol secretion and the hepatic gene expression of ABCG5/G8. 
In paper II, it was investigated if the TH-induced secretion of cholesterol into bile is mediated 
by ABCG5/G8, or if other pathways are involved. TH-induced secretion of cholesterol into bile 
was found to be largely dependent on the ABCG5/G8 transporter. It was also found that nuclear 
hormone receptor LXRα, reported to be positively regulated by TH and to have a stimulatory 
effect on ABCG5/G8 gene expression, is not critical for the TH-induced effect on ABCG5/G8 
gene expression or on biliary cholesterol secretion.



  
In paper III, the responses to hyperthyroidism in humans were studied and compared to those of 
healthy subjects treated with the liver-specific thyromimetic eprotirome. Hyperthyroidism 
lowered VLDL-, LDL-, and HDL-cholesterol, apoB, Lp(a), and serum PCSK9. Bile acid 
synthesis was increased and serum FGF19 reduced. Cholesterol synthesis was unaltered while 
intestinal absorption of dietary cholesterol was reduced. Serum free fatty acids and glycerol 
were increased, while insulin, glucose, and FGF21 were unaltered. Eprotirome treatment 
resulted in similar reductions in lipoprotein cholesterol, apoB, Lp(a), and PCSK9. In contrast to 
hyperthyroidism, eprotirome reduced plasma triglycerides. There were no effects on bile acid 
synthesis, FGF19, or cholesterol absorption, in response to liver-selective stimulation of TH 
receptors by eprotirom
Glycogen synthase kinase 3-mediated voltage-dependent anion channel phosphorylation controls outer mitochondrial membrane permeability during lipid accumulation.

International audienceUNLABELLED: Nonalcoholic steatosis is a liver pathology characterized by fat accumulation and severe metabolic alterations involving early mitochondrial impairment and late hepatocyte cell death. However, mitochondrial dysfunction mechanisms remain elusive. Using four models of nonalcoholic steatosis, i.e., livers from patients with fatty liver disease, ob/ob mice, mice fed a high-fat diet, and in vitro models of lipotoxicity, we show that outer mitochondrial membrane permeability is altered and identified a posttranslational modification of voltage-dependent anion channel (VDAC), a membrane channel and NADH oxidase, as a cause of early mitochondrial dysfunction. Thus, in nonalcoholic steatosis VDAC exhibits reduced threonine phosphorylation, which increases the influx of water and calcium into mitochondria, sensitizes the organelle to matrix swelling, depolarization, and cytochrome c release without inducing cell death. This also amplifies VDAC enzymatic and channel activities regulation by calcium and modifies its interaction with proteic partners. Moreover, lipid accumulation triggers a rapid lack of VDAC phosphorylation by glycogen synthase kinase 3 (GSK3). Pharmacological and genetic manipulations proved GSK3 to be responsible for VDAC phosphorylation in normal cells. Notably, VDAC phosphorylation level correlated with steatosis severity in patients. CONCLUSION: VDAC acts as an early sensor of lipid toxicity and its GSK3-mediated phosphorylation status controls outer mitochondrial membrane permeabilization in hepatosteatosis
A randomized controlled trial of pretransplant antiviral therapy to prevent recurrence of hepatitis C after liver transplantation

Hepatitis C virus (HCV) infection recurs in liver recipients who are viremic at transplantation. We conducted a randomized, controlled trial to test the efficacy and safety of pretransplant pegylated interferon alpha‐2b plus ribavirin (Peg‐IFN‐α2b/RBV) for prevention of post‐transplant HCV recurrence. Enrollees had HCV and were listed for liver transplantation, with either potential living donors or Model for End‐Stage Liver Disease upgrade for hepatocellular carcinoma. Patients with HCV genotypes (G) 1/4/6 (n = 44/2/1) were randomized 2:1 to treatment (n = 31) or untreated control (n = 16); HCV G2/3 (n=32) were assigned to treatment. Overall, 59 were treated and 20 were not. Peg‐IFN‐α2b, starting at 0.75 μg/kg/week, and RBV, starting at 600 mg/day, were escalated as tolerated. Patients assigned to treatment versus control had similar baseline characteristics. Combined virologic response (CVR) included pretransplant sustained virologic response and post‐transplant virologic response (pTVR), defined as undetectable HCV RNA 12 weeks after end of treatment or transplant, respectively. In intent‐to‐treat analyses, 12 (19%) assigned to treatment and 1 (6%) assigned to control achieved CVR ( P  = 0.29); per‐protocol values were 13 (22%) and 0 (0%) ( P  = 0.03). Among treated G1/4/6 patients, 23 of 30 received transplant, of whom 22% had pTVR; among treated G2/3 patients 21 of 29 received transplant, of whom 29% had pTVR. pTVR was 0%, 18%, and 50% in patients treated for <8, 8‐16, and >16 weeks, respectively ( P  = 0.01). Serious adverse events (SAEs) occurred with similar frequency in treated versus untreated patients (68% versus 55%;  P  = 0.30), but the number of SAEs per patient was higher in the treated group (2.7 versus 1.3;  P  = 0.003).  Conclusion : Pretransplant treatment with Peg‐IFN‐α2b/RBV prevents post‐transplant recurrence of HCV in selected patients. Efficacy is higher with >16 weeks of treatment, but treatment is associated with increased risk of potentially serious complications. (H EPATOLOGY  2013
Determination of reliability criteria for liver stiffness evaluation by transient elastography

UNLABELLED: Liver stiffness evaluation (LSE) is usually considered as reliable when it fulfills all the following criteria: ≥10 valid measurements, ≥60% success rate, and interquartile range / median ratio (IQR/M) ≤0.30. However, such reliable LSE have never been shown to be more accurate than unreliable LSE. Thus, we aimed to evaluate the relevance of the usual definition for LSE reliability, and to improve reliability by using diagnostic accuracy as a primary outcome in a large population. 1,165 patients with chronic liver disease from 19 French centers were included. All patients had liver biopsy and LSE. 75.7% of LSE were reliable according to the usual definition. However, these reliable LSE were not significantly more accurate than unreliable LSE with, respectively: 85.8% versus 81.5% well-classified patients for the diagnosis of cirrhosis (P = 0.082). In multivariate analyses with different diagnostic targets, LSE median and IQR/M were independent predictors of fibrosis staging, with no significant influence of ≥10 valid measurements or LSE success rate. These two reliability criteria determined three LSE groups: "very reliable" (IQR/M ≤0.10), "reliable" (0.10&lt; IQR/M ≤0.30, or IQR/M &gt;0.30 with LSE median &lt;7.1 kPa), and "poorly reliable" (IQR/M &gt;0.30 with LSE median ≥7.1 kPa). The rates of well-classified patients for the diagnosis of cirrhosis were, respectively: 90.4%, 85.8%, and 69.5% (P &lt; 10(-3) ). According to these new reliability criteria, 9.1% of LSE were poorly reliable (versus 24.3% unreliable LSE with the usual definition, P &lt; 10(-3) ), 74.3% were reliable, and 16.6% were very reliable.
CONCLUSION: The usual definition for LSE reliability is not relevant. LSE reliability depends on IQR/M according to liver stiffness median level, defining thus three reliability categories: very reliable, reliable, and poorly reliable LSE. (HEPATOLOGY 2013)
Chimeric hepatitis B virus/hepatitis C virus envelope proteins elicit broadly neutralizing antibodies and constitute a potential bivalent prophylactic vaccine.

International audienceThe development of a prophylactic vaccine against hepatitis C virus (HCV) has become an important medical priority, because 3-4 million new HCV infections are thought to occur each year worldwide. Hepatitis B virus (HBV) is another major human pathogen, but infections with this virus can be prevented with a safe, efficient vaccine, based on the remarkable ability of the envelope protein (S) of this virus to self-assemble into highly immunogenic subviral particles. Chimeric HBV-HCV envelope proteins in which the N-terminal transmembrane domain of S was replaced with the transmembrane domain of the HCV envelope proteins (E1 or E2) were efficiently coassembled with the wild-type HBV S protein into subviral particles. These chimeric particles presented the full-length E1 and E2 proteins from a genotype 1a virus in an appropriate conformation for formation of the E1-E2 heterodimer. Produced in stably transduced Chinese hamster ovary cells and used to immunize New Zealand rabbits, these particles induced a strong specific antibody (Ab) response against the HCV and HBV envelope proteins in immunized animals. Sera containing anti-E1 or anti-E2 Abs elicited by these particles neutralized infections with HCV pseudoparticles and cell-cultured viruses derived from different heterologous 1a, 1b, 2a, and 3 strains. Moreover, the anti-hepatitis B surface response induced by these chimeric particles was equivalent to the response induced by a commercial HBV vaccine. Conclusions: Our results provide support for approaches based on the development of bivalent HBV-HCV prophylactic vaccine candidates potentially able to prevent initial infection with either of these two hepatotropic viruses. (HEPATOLOGY 2013)
Analysis of hepatitis C virus resistance to silibinin in vitro and in vivo points to a novel mechanism involving nonstructural protein 4B.

Intravenous silibinin (SIL) is an approved therapeutic that has recently been applied to patients with chronic hepatitis C, successfully clearing hepatitis C virus (HCV) infection in some patients even in monotherapy. Previous studies suggested multiple antiviral mechanisms of SIL; however, the dominant mode of action has not been determined. We first analyzed the impact of SIL on replication of subgenomic replicons from different HCV genotypes in vitro and found a strong inhibition of RNA replication for genotype 1a and genotype 1b. In contrast, RNA replication and infection of genotype 2a were minimally affected by SIL. To identify the viral target of SIL we analyzed resistance to SIL in vitro and in vivo. Selection for drug resistance in cell culture identified a mutation in HCV nonstructural protein (NS) 4B conferring partial resistance to SIL. This was corroborated by sequence analyses of HCV from a liver transplant recipient experiencing viral breakthrough under SIL monotherapy. Again, we identified distinct mutations affecting highly conserved amino acid residues within NS4B, which mediated phenotypic SIL resistance also in vitro. Analyses of chimeric viral genomes suggest that SIL might target an interaction between NS4B and NS3/4A. Ultrastructural studies revealed changes in the morphology of viral membrane alterations upon SIL treatment of a susceptible genotype 1b isolate, but not of a resistant NS4B mutant or genotype 2a, indicating that SIL might interfere with the formation of HCV replication sites. CONCLUSION: Mutations conferring partial resistance to SIL treatment in vivo and in cell culture argue for a mechanism involving NS4B. This novel mode of action renders SIL an attractive candidate for combination therapies with other directly acting antiviral drugs, particularly in difficult-to-treat patient cohorts