Obesity dependent metabolic signatures associated with nonalcoholic fatty liver disease progression

Our understanding of the mechanisms by which nonalcoholic fatty liver disease (NAFLD) progresses from simple steatosis to steatohepatitis (NASH) is still very limited. Despite the growing number of studies linking the disease with altered serum metabolite levels, an obstacle to the development of metabolome-based NAFLD predictors has been the lack of large cohort data from biopsy-proven patients matched for key metabolic features such as obesity. We studied 467 biopsied individuals with normal liver histology (n=90) or diagnosed with NAFLD (steatosis, n=246; NASH, n=131), randomly divided into estimation (80% of all patients) and validation (20% of all patients) groups. Qualitative determinations of 540 serum metabolite variables were performed using ultra-performance liquid chromatography coupled to mass spectrometry (UPLCMS). The metabolic profile was dependent on patient body-mass index (BMI), suggesting that the NAFLD pathogenesis mechanism may be quite different depending on an individual’s level of obesity. A BMI-stratified multivariate model based on the NAFLD serum metabolic profile was used to separate patients with and without NASH. The area under the receiver operating characteristic curve was 0.87 in the estimation and 0.85 in the validation group. The cutoff (0.54) corresponding to maximum average diagnostic accuracy (0.82) predicted NASH with a sensitivity of 0.71 and a specificity of 0.92 (negative/positive predictive values = 0.82/0.84). The present data, indicating that a BMI-dependent serum metabolic profile may be able to reliably distinguish NASH from steatosis patients, have significant implications for the development of NASH biomarkers and potential novel targets for therapeutic intervention

Relevance of p53 in the regulation of pro and antiapoptotic factors from the Bcl-2 family during the treatment with tirosine kinase inhibitors

Motivation: Liver cancer is the sixth most common cancer and the fourth most frequent cause of cancer-related death worldwide [1]. Nowadays, only one third of patients diagnosed with HCC are in the earliest BCLC 0-A stages, with a high proportion being in more advanced stages of the disease (BCLC B-C). Patients with the presence of poor prognostic factors such as vascular invasion, extrahepatic metastases and/or impaired hepatic function are considered in the advanced stage of the disease (BCLC C). Sorafenib is the standard of care for advanced HCC stage demonstrated in two large-scale trials [2]. Other drugs have been developed to increase the therapeutic arsenal of treatments. A phase III clinical trial comparing Lenvatinib and Sorafenib demonstrated that Lenvatinib was statistically non-inferior to Sorafenib in overall survival as a first-line treatment in patients with advanced HCC. Regorafenib and Cabozantinib have been shown to be effective as second-line therapy. [2]   Methods: Sorafenib, Regorafenib, Cabozantinib and Lenvatinib were obtained commercially from Carbosynth Limited (Berkshire, UK). HepG2, Hep3B and Huh7 cell lines were obtained from American Type Culture Collection (ATCC; LGC Standards, S.L.U., Barcelona, Spain). HCC cell lines were maintained in supplemented Minimum Essential Medium with Earle's Balanced Salts (MEM/EBSS) at 37°C in a humidified incubator with 5 % CO2. Cells were seeded at a density of 100,000 cells/cm2 in 2D culture. Different parameters related to cell death and proliferation were associated with the expression of proapoptotic (Bak, Bax, tBid and Bim) antiapoptotic (Bcl-2, Mcl-1 and Bcl-xL) and regulatory (Beclin-1) Bcl-2 family members assessed by Western-blot analysis.   Results: The administration of tyrosine kinase inhibitors induced cell death and reduced cell proliferation. This effect was associated with an upregulation of tBid and Bim expression in liver cancer cells. This effect was not observed in Hep3B and Huh7 which were less responsiveness to the proapoptotic and antiproliferative properties of tyrosine kinase inhibitors.   Conclusions: The induction of cell death and antiproliferative properties of tyrosine kinase inhibitors were associated with the increase of the expression of different proapoptotic Bcl-2 family members. This expression appeared to be regulated by p53

Hepatic changes during a carrageenan induced granuloma in rats

Hepatic changes during inflammation were studied in rats bearing a carrageenan induced granuloma. In spite of a decrease in the metabolic capacity of microsomes to induce lipid peroxidation during inflammation, the endogenous lipid peroxidation remained unchanged and unrelated with the hepatic activities measured. The continuous increase in hepatic cAMP observed during acute and chronic phases could be related to adenylate cyclase stimulation by mediators, and could be an initial step in the hepatocyte adaptation leading to the increased level of hepatic caeruloplasmin, to the reduction of cytochrome P-450 level and to the modifications of Ca2+ sequestration by microsomes

L'usage de l'espace par les exploitations d'élevage de montagne et la gestion de la biodiversité

International audienceEn prenant appui sur divers résultats de recherches menées depuis 20 ans dans les Pyrénées centrales, les caractéristiques de l'utilisation de l'espace par les exploitations agricoles sont présentées. Les particularités spatiales des territoires d'exploitation influent sur les pratiques d'utilisation des prairies et sur la dynamique des couverts et des paysages. Un modèle de référence permet de diagnostiquer la maîtrise de l'exploitation de la végétation. L'impact des modes d'usage des prairies sur leur richesse en espèces est présenté sur quelques cas concrets. Une gestion mal maîtrisée conduit à une augmentation de la diversité intraparcellaire et à une diminution de la diversité interparcellaire, puis à une baisse rapide de ces 2 diversités. Raisonner l'organisation de l'usage des prairies à des niveaux d'organisation larges est une nécessité pour préserver la biodiversité en région de montagne

The search for novel diagnostic and prognostic biomarkers in cholangiocarcinoma

[EN] The poor prognosis of cholangiocarcinoma (CCA) is in part due to late diagnosis, which is currently achieved by a combination of clinical, radiological and histological approaches. Available biomarkers determined in serum and biopsy samples to assist in CCA diagnosis are not sufficiently sensitive and specific. Therefore, the identification of new biomarkers, preferably those obtained by minimally invasive methods, such as liquid biopsy, is important. The development of innovative technologies has permitted to identify a significant number of genetic, epigenetic, proteomic and metabolomic CCA features with potential clinical usefulness in early diagnosis, prognosis or prediction of treatment response. Potential new candidates must be rigorously evaluated prior to entering routine clinical application. Unfortunately, to date, no such biomarker has achieved validation for these purposes. This review is an up-to-date of currently used biomarkers and the candidates with promising characteristics that could be included in the clinical practice in the next future. This article is part of a Special Issue entitled: Cholangiocytes in Health and Disease edited by Jesus Banales, Marco Marzioni, Nicholas LaRusso and Peter Jansen

Effects of a hyperlipidic diet on experimental carcinogenesis of the breast: content and type of tumors

En el presente trabajo se han estudiado los efectos de una dieta hiperlipídica (20 % aceite de maíz) sobre las principales etapas de la carcinogénesis. Dicho estudio se ha realizado utilizando como soporte experimental el modelo del cáncer de mama inducido en la rata mediante dimetilbenz(a) antraceno. Los tumores mamarios malignos representan la patología más abundante aparecida en los animales. Tal resultado es atribuido específicamente a la acción del carcinógeno. Por otra parte, al comparar el número de tales tumores entre los diferentes grupos experimentales y el grupo control, se observa que la dieta hiperlipídica administrada tiene efectos promotores de la carcinogénesis mamaria en la rata. Este efecto no se obtiene si la dieta hiperlipídica se administra a los 157 días de la inducción carcinogénica. Además, los resultados obtenidos también indican que dicha dieta no tiene acción sobre la iniciación. Sin embargo, tal probabilidad no puede afirmarse dado que cabe la posibilidad de que la dieta hiperlipídica haya inducido cambios en los animales que hayan modificado la eficacia de la carcinogénesis

Impaired autophagic flux is associated with increased endoplasmic reticulum stress during the development of NAFLD

This work is licensed under a Creative Commons Attribution-NonCommercialNoDerivs 3.0 Unported License.-- et al.The pathogenic mechanisms underlying the progression of non-alcoholic fatty liver disease (NAFLD) are not fully understood. In this study, we aimed to assess the relationship between endoplasmic reticulum (ER) stress and autophagy in human and mouse hepatocytes during NAFLD. ER stress and autophagy markers were analyzed in livers from patients with biopsy-proven non-alcoholic steatosis (NAS) or non-alcoholic steatohepatitis (NASH) compared with livers from subjects with histologically normal liver, in livers from mice fed with chow diet (CHD) compared with mice fed with high fat diet (HFD) or methionine-choline-deficient (MCD) diet and in primary and Huh7 human hepatocytes loaded with palmitic acid (PA). In NASH patients, significant increases in hepatic messenger RNA levels of markers of ER stress (activating transcription factor 4 (ATF4), glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP)) and autophagy (BCN1) were found compared with NAS patients. Likewise, protein levels of GRP78, CHOP and p62/SQSTM1 (p62) autophagic substrate were significantly elevated in NASH compared with NAS patients. In livers from mice fed with HFD or MCD, ER stress-mediated signaling was parallel to the blockade of the autophagic flux assessed by increases in p62, microtubule-associated protein 2 light chain 3 (LC3-II)/LC3-I ratio and accumulation of autophagosomes compared with CHD fed mice. In Huh7 hepatic cells, treatment with PA for 8 h triggered activation of both unfolding protein response and the autophagic flux. Conversely, prolonged treatment with PA (24 h) induced ER stress and cell death together with a blockade of the autophagic flux. Under these conditions, cotreatment with rapamycin or CHOP silencing ameliorated these effects and decreased apoptosis. Our results demonstrated that the autophagic flux is impaired in the liver from both NAFLD patients and murine models of NAFLD, as well as in lipid-overloaded human hepatocytes, and it could be due to elevated ER stress leading to apoptosis. Consequently, therapies aimed to restore the autophagic flux might attenuate or prevent the progression of NAFLD.We acknowledge the following grant support: SAF2012-33283 (MINECO, Spain), Comunidad de Madrid S2010/BMD-2423, EFSD and Amylin Paul Langerhans Grant and Centro de Investigación Biomédica en Red de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM, ISCIII, Barcelona, Spain) to AMV.; SAF2010-16037, SAF2013-43713-R (MINECO) and Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBEREHD, ISCIII) to PMS. RD12/0042/0019 (ISCIII) and S2010/BMD-2478 (Comunidad de Madrid) to LB, PI 13/01299 and Fundación Mutua Madrileña 2012 to C G-M and AIRC IG-2012 to GMF.Peer Reviewe

DDR1 and Its Ligand, Collagen IV, Are Involved in In Vitro Oligodendrocyte Maturation

Discoidin domain receptor 1 (DDR1) is a tyrosine kinase receptor expressed in epithelial cells from different tissues in which collagen binding activates pleiotropic functions. In the brain, DDR1 is mainly expressed in oligodendrocytes (OLs), the function of which is unclear. Whether collagen can activate DDR1 in OLs has not been studied. Here, we assessed the expression of DDR1 during in vitro OL differentiation, including collagen IV incubation, and the capability of collagen IV to induce DDR1 phosphorylation. Experiments were performed using two in vitro models of OL differentiation: OLs derived from adult rat neural stem cells (NSCs) and the HOG16 human oligodendroglial cell line. Immunocytofluorescence, western blotting, and ELISA were performed to analyze these questions. The differentiation of OLs from NSCs was addressed using oligodendrocyte transcription factor 2 (Olig2) and myelin basic protein (MBP). In HOG16 OLs, collagen IV induced DDR1 phosphorylation through slow and sustained kinetics. In NSC-derived OLs, DDR1 was found in a high proportion of differentiating cells (MBP+/Olig2+), but its protein expression was decreased in later stages. The addition of collagen IV did not change the number of DDR1+/MBP+ cells but did accelerate OL branching. Here, we provide the first demonstration that collagen IV mediates the phosphorylation of DDR1 in HOG16 cells and that the in vitro co-expression of DDR1 and MBP is associated with accelerated branching during the differentiation of primary OLs

DDR1 and Its Ligand, Collagen IV, Are Involved in In Vitro Oligodendrocyte Maturation

Discoidin domain receptor 1 (DDR1) is a tyrosine kinase receptor expressed in epithelial cells from different tissues in which collagen binding activates pleiotropic functions. In the brain, DDR1 is mainly expressed in oligodendrocytes (OLs), the function of which is unclear. Whether collagen can activate DDR1 in OLs has not been studied. Here, we assessed the expression of DDR1 during in vitro OL differentiation, including collagen IV incubation, and the capability of collagen IV to induce DDR1 phosphorylation. Experiments were performed using two in vitro models of OL differentiation: OLs derived from adult rat neural stem cells (NSCs) and the HOG16 human oligodendroglial cell line. Immunocytofluorescence, western blotting, and ELISA were performed to analyze these questions. The differentiation of OLs from NSCs was addressed using oligodendrocyte transcription factor 2 (Olig2) and myelin basic protein (MBP). In HOG16 OLs, collagen IV induced DDR1 phosphorylation through slow and sustained kinetics. In NSC-derived OLs, DDR1 was found in a high proportion of differentiating cells (MBP+/Olig2+), but its protein expression was decreased in later stages. The addition of collagen IV did not change the number of DDR1+/MBP+ cells but did accelerate OL branching. Here, we provide the first demonstration that collagen IV mediates the phosphorylation of DDR1 in HOG16 cells and that the in vitro co-expression of DDR1 and MBP is associated with accelerated branching during the differentiation of primary OLs