Human and experimental evidence supporting a role for osteopontin in alcoholic hepatitis: Hepatology

We identified in the transcriptome analysis of patients with alcoholic hepatitis (AH) osteopontin (OPN) as one of the most up-regulated genes. Here, we used a translational approach to investigate its pathogenic role. OPN hepatic gene expression was quantified in patients with AH and other liver diseases. OPN protein expression and processing were assessed by immmunohistochemistry, Western blotting and ELISA. OPN gene polymorphisms were evaluated in patients with alcoholic liver disease. The role of OPN was evaluated in OPN−/− mice with alcohol-induced liver injury. OPN biological actions were studied in human hepatic stellate cells and in precision-cut liver slices. Hepatic expression and serum levels of OPN were markedly increased in AH compared to normal livers and other types of chronic liver diseases and correlated with short-term survival. Serum levels of OPN also correlated with hepatic expression and disease severity. OPN was mainly expressed in areas with inflammation and fibrosis. Two proteases that process OPN (thrombin and MMP-7) and cleaved-OPN were increased in livers with AH. Patients with AH had a tendency of a lower frequency of the CC genotype of the +1239C SNP of the OPN gene compared to patients with alcohol abuse without liver disease. Importantly, OPN−/− mice were protected against alcohol-induced liver injury and showed decreased expression of inflammatory cytokines. Finally, OPN was induced by LPS and stimulated inflammatory actions in hepatic stellate cells

Vulnérabilité génétique à l'alcoolodépendance (du knock-out à la pharmacogénétique)

Les avancées récentes dans le domaine des neurosciences et de la génétique ont permis une meilleure compréhension de l influence génétique sur les réponses liées à l alcool et la vulnérabilité différentielle à l alcoolodépendance au niveau cellulaire et moléculaire. Cependant, l efficacité modeste des pharmacothérapies utilisées dans le traitement de cette maladie chronique et hautement récidivante nécessite la recherche de nouveaux traitements et/ou un choix du traitement en fonction du patrimoine génétique des malades. C est dans ce contexte que le présent travail de thèse s est intéressé au rôle de deux systèmes neuromodulateurs (endocannabinoïdergique et adénosinergique) dans l alcoolodépendance ainsi qu aux facteurs génétiques associés avec différents traits de la maladie et/ou aux dommages somatiques. Enfin ce travail a également concerné, pour la première fois au niveau mondial l étude de la réponse aux deux traitements actuellement préconisés en fonction de polymorphismes de gènes candidats. Les résultats ont démontré que les deux systèmes neuromodulateurs étudiés jouent un rôle important dans différentes composantes de la maladie et ouvrent des perspectives intéressantes en termes de nouveaux traitements visant le système adénosinergique. Les résultats obtenus dans les études cliniques ont permis d identifier des associations de certains polymorphismes avec la sévérité de la maladie ainsi qu avec la vulnérabilité à développer une maladie alcoolique du foie. Enfin, notre étude européenne a permis de démontrer clairement l intérêt d utiliser les tests génétiques dans la prédiction de l efficacité des traitements de la dépendance à l alcool.Recent advances in neuroscience and genetics have enabled a better understanding of genetically influenced differences in ethanol ( alcohol )-related responses and differential vulnerability to alcohol dependence at the cellular and molecular levels. However, the poor pharmacotherapy effectiveness regarding this chronic and highly relapsing disorder requires the research of new pharmacological treatments and/or the choice of the treatment depending on the gene variants of the patient. In this context, the present thesis work aimed at identifying the role of two neuromodulator systems (i.e. endocannabinoidergic and adenosinergic) in alcohol dependence and also the genetic factors associated with several traits of the disease and/or its somatic damages. Finally, this work aimed, for the first time at the international level, at analyzing the response to both treatments actually used in alcohol dependence, depending on gene variants of candidate genes. The results demonstrated that both neuromodulator systems play an important role in different components of the disease and that the adenosine system represents a promising therapeutic avenue. The results of clinical studies revealed associations of some gene variants with the severity of alcohol dependence and vulnerability to alcoholic liver disease. Finally, our European study clearly demonstrated the interest of using genetic tests in predicting the effectiveness of treatments of alcohol dependence.AMIENS-BU Santé (800212102) / SudocPARIS-BIUP (751062107) / SudocSudocFranceF

Le polymorphisme génétique de la PNPLA 3 dans l'hépatite alcoolique aiguë sévère

Introduction : Le polymorphisme génétique de la Patatin-like phospholipase domain-containing protein 3 (PNPLA3) est associé à la cirrhose d origine alcoolique. Le but de notre étude a été d étudier l impact du polymorphisme de la PNPLA3 dans l hépatite alcoolique aiguë sévère (HAA). Patients et méthodes : Le polymorphisme rs738409 C>G de la PNPLA3 a été étudié chez 208 personnes non apparentées, comprenant 64 patients ayant une HAA, 39 patients avec une cirrhose sans HAA et 105 témoins sains appariés sur l âge et le sexe. Les fréquences génotypiques et allèliques étaient comparées entre les groupes. Les caractéristiques cliniques et biologiques des différents génotypes étaient analysées dans le groupe HAA. Résultats : Les patients du groupe HAA avaient une fonction hépatique plus altérée que celle des patients du groupe Cirrhose (MELD 24,5 versus 14, p G de la PNPLA3 est associé à la cirrhose alcoolique et à l hépatite alcoolique aiguë sévère. L allèle muté G est un facteur indépendant de mortalité chez les patients ayant une hépatite alcoolique aiguë sévère.Background : The Patatin-like phospholipase domain-containing protein 3 (PNPLA 3) polymorphism is associated with alcoholic cirrhosis. The purpose of our study was to investigate the impact of the PNPLA 3 gene polymorphism in severe acute alcoholic hepatitis (AAH). Patients and Methods : The PNPLA 3 rs738409 polymorphism was studied in 208 non relatives patients, including 64 AAH patients, 39 alcoholic cirrhosis patients without AAH and 105 controls matched for age and sex. Genotypic and allelic frequencies were compared between groups. Clinical and biological characteristics of the different genotypes were analyzed in the AAH group. Results : Compared to cirrhosis group, patients from the AAH group had more impaired liver function (MELD 24,5 versus 14, pG polymorphism of the PNPLA 3 gene is associated with alcoholic cirrhosis and severe acute alcoholic hepatitis. The mutant G allele is an independent predictive factor of mortality in patients with AAH.AMIENS-BU Santé (800212102) / SudocSudocFranceF

Etat des lieux en Picardie, de la pratique du repérage précoce de l'usage abusif d'alcool en médecine générale

Avec près de 45.000 décès annuels attribuables à l alcool, l inquiétante prévalence du mésusage devrait faire l objet d attentions particulières de la part des instances sanitaires en France. La prévention des modes d usages inadaptés de l alcool bénéficie aujourd hui d outils de diagnostic qui sont notamment dédiés à la pratique en médecine générale. Cependant l utilisation de ces outils nécessite la connaissance des définitions des différents usages de l alcool et des repères standardisés proposés par les autorités sanitaires françaises. Or le constat au niveau national montre des lacunes manifestes qui n épargnent pas la Picardie. La place centrale du médecin généraliste au sein des soins primaires fait de lui un acteur incontournable pour élaborer une stratégie de systématisation du repérage du mésusage d alcool, qui est, sans aucun doute, la réponse la plus efficace pour lutter contre l inquiétante prévalence de l abus d alcool avec ou sans dépendance. Si les lacunes des médecins généralistes sont bien identifiées, la question de savoir s ils sont prêts à bénéficier d une formation qui leurs apporterait les connaissances nécessaires n a pas encore été posée en Picardie A l aide d un questionnaire adressé à 420 médecins généralistes de la Picardie, nous avons sondé l opinion et les connaissances de ces derniers pour déterminer quel accueil pouvait être fait à la formation au Repérage Précoce et à l Intervention Brève (RPIB). L analyse des réponses a effectivement confirmé les faiblesses des connaissances de base en alcoologie, mais a surtout montré par une large majorité que les médecins généralistes accueillent de manière favorable la formation et accepteraient la notion de systématisation du repérage du mésusage. La conclusion de ce travail a pour perspective de restituer nos résultats à tous les médecins généralistes de la Picardie dans le but de les inciter à participer aux formations au RPIB.In France, more than 45 thousands of human death reliable to alcohol abuse must lead to give more attentions to the strategic project against the alcohol misuse prevalence. Diagnostic tools directed to alcohol misuse are currently adapted to general practitioners. However, to correctly use these tools a general practitioner needs to display a good knowledge of all misuse definitions and thresholds recommendations of alcohol use. Unfortunately, a dramatic national observation showed the lake of knowledge of basic definition in Picardie and in France. The strategic place of the general practitioner in the center of the primary care gives him the best role to build systematisation strategies to alcohol abuse screening. Lakes of basic definitions within general practitioners are commonly admitted, however, in Picardie the question to general practitioners has been never asked to know how they could receive the learning of RPIB practice. Using a questionnaire send to 420 general practitioners in whole Picardie, we tried to evaluate the knowledge of definition and how general practitioners could appreciate an adapted learning to RPIB. Result of this work confirmed the lake of knowledge, but it is noteworthy that general practitioners give a majority of a good welcome to RPIB learning and to the systematisation of this practice. The work could be concluding with a nice perspective directed to resituate the result of this survey to initiate a greater participation to RPIB learning.AMIENS-BU Santé (800212102) / SudocSudocFranceF

The genetics of alcoholic liver disease: better patient group definition is required.

International audienc

Binge Eating, But Not Other Disordered Eating Symptoms, Is a Significant Contributor of Binge Drinking Severity: Findings from a Cross-Sectional Study among French Students

International audienc

The -308 TNFalpha gene polymorphism in severe acute alcoholic hepatitis: identification of a new susceptibility marker.

International audienceBACKGROUND: This study investigated whether genetic polymorphisms in the tumor necrosis factor alpha (TNFalpha) gene's promoter region play a role in severe acute alcoholic hepatitis (AAH). METHODS: One-hundred and fifty patients (58 AAH patients, 45 cirrhosis group free-AAH, 47 healthy group) were genotyped for 3 TNFalpha polymorphisms (-238, -308, -863) using a polymerase chain reaction-restriction fragment length polymorphism technique. Serum TNFalpha levels were determined. RESULTS: The TNFalpha-308 allele A frequency was significantly lower in AAH group (0.09), than cirrhosis (0.28) (p < 0.001), and healthy groups (p < 0.001). The TNFalpha-308 A/G, A/A genotypes were significantly lower in AAH group, than cirrhosis (p = 0.005), and healthy groups (p < 0.001). For AAH group, there were no clinical, biological, and serum TNFalpha differences between the -308 G/G and A/G, A/A genotype patients, apart higher transaminase in the former group (p = 0.02). AAH and cirrhosis groups did not differ for the frequency of TNFalpha-238, -863 polymorphisms. The specific genotype did not appear to have any influence on the therapeutic response following corticotherapy or posttreatment 6-month survival. In the AAH group, nonsurvivors had higher TNFalpha levels than survivors (7.9 +/- 8.8 vs. 3.3 +/- 1.6 pg/ml, p = 0.01). CONCLUSIONS: These results attest to the involvement of TNFalpha-related genetic factors in susceptibility to AAH

Lack of association between tumour necrosis factor receptor types 1 and 2 gene polymorphism and severe acute alcoholic hepatitis.

International audienceBACKGROUND/AIMS: Tumour necrosis factor-alpha (TNF-alpha) is involved in the physiopathology of severe acute alcoholic hepatitis (AAH) binding with TNF receptor types TNFR1 and TNFR2, whose serum concentrations are elevated. We studied the role of TNFR1 and TNFR2 gene polymorphism in AAH patients. METHODS: One hundred and ninety-two patients (58 AAH with Maddrey score >or=32, 44 non-AAH cirrhoses, 90 healthy individuals) were genotyped for A36G TNFR1 and T676G TNFR2 using polymerase chain reaction-restriction fragment length polymorphism technique. Serum sTNFR1 and sTNFR2 were assayed. RESULTS: The AAH and two control groups did not differ for genotype distribution. In three groups, A (36 TNFR1) and T (676 TNFR2) allelic frequencies were similar, at 0.47, 0.47, 0.44 and 0.78, 0.81, 0.80, respectively. The 36 TNFR1, 676 TFNR2 genotypes did not influence on prognostic scores (Maddrey, Child-Pugh), nor in response to corticosteroids or 6-month survival. sTNFR1 levels were higher in AAH than healthy group (3.07+/-1.14 vs. 1.17+/-0.27 ng/ml, P<0.001) and sTNFR2 levels were higher in AAH than cirrhosis (3.6+/-1.02 vs. 3.1+/-1.03, P<0.05) and healthy groups (3.6+/-1.02 vs. 1.91+/-0.54, P<0.001). However, sTNFR1 and sTNFR2 levels did not vary significantly according to genotypes. CONCLUSION: These results did not support an association between 36 TNFR1, 676 TNFR2 gene polymorphisms and AAH

Effects of prenatal and postnatal maternal ethanol on offspring response to alcohol and psychostimulants in long evans rats.

International audienceAn important factor that may influence addiction liability is exposure during the early life period. Exposure to ethanol, early in life, can have long-lasting implications on brain function and drugs of abuse response later in life. In the present study we investigated the behavioral responses to ethanol and to psychostimulants in Long Evans rats that have been exposed to pre- and postnatal ethanol. Since a relationship between heightened drug intake and susceptibility to drug-induced locomotor activity/sensitization has been demonstrated, we tested these behavioral responses, in control and early life ethanol-exposed animals. The young adult male and female progeny were tested for locomotor response to alcohol, cocaine and d-amphetamine. Sedative, rewarding effects of alcohol and alcohol consumption were measured. Our results show that early life ethanol exposure behaviorally sensitized animals to subsequent ethanol and psychostimulants exposure. Ethanol-exposed animals were also more sensitive to the hyperlocomotor effects of all drugs of abuse tested and to those of the dopamine receptor agonist apomorphine. Locomotor sensitization to repeated injections of cocaine was facilitated in ethanol-exposed animals. Ethanol-induced conditioned place preference was also facilitated in ethanol-exposed animals. Ethanol consumption and preference were increased after early life ethanol exposure and this was associated with decreased sensitivity to the sedative effects of ethanol. The altered behavioral responses to drugs of abuse were associated with decreased striatal dopamine transporter and hippocampal NMDAR binding. Our results outline an increased vulnerability to rewarding and stimulant effects of ethanol and psychostimulants and support the epidemiological and clinical data that suggested that early chronic exposure to ethanol may increase the propensity for later self-administration of ethanol or other substances