The current winners of the previous 2021 annual meeting were:
CTPIOD basic research 1st prize to Jorge Garcia Garcia
2nd prize to Elisa Felix Soriano
CTPIOD clinic research 1st prize to Marta Lopez Yus
Below their pictures is listed in alphebetical order the abstracts participating to the contest
CTPIOD basic research 1st prize to Jorge Garcia Garcia
2nd prize to Elisa Felix Soriano
CTPIOD clinic research 1st prize to Marta Lopez Yus
Below their pictures is listed in alphebetical order the abstracts participating to the contest
LIST OF THE ABSTRACTS EXAMINED FOR THE RESEARCH AWARDS IN 2021:
Early alteration of the adipose tissue adaptive response to fasting/refeeding under a high fat diet:
sex and depot specificity
Alex Aupetit, Pauline Decaunes, Kévin Perrier, Pascale Loubière, Rémy Flores-Flores Anne Bouloumié, Anaïs Briot
Institute of Metabolic and Cardiovascular Diseases, INSERM, UMR1297, Team 1, I2MC, CHU Rangueil,
BP84225, 1 avenue Jean Poulhès, 31432, Toulouse cedex 4, France.
Metabolic adaptation to fasting and refeeding is critical to maintain cardiometabolic health. The white adipose tissue (AT) plays a central role in the maintenance of this adaptation. In post-prandial condition fatty acids in excess are stored under the form of triglycerides (lipogenesis) into mature adipocytes while they are released (lipolysis) during prolonged fasting. Disruption of AT function, as observed with obesity and aging, contributes to the development of chronic diseases such as type 2 diabetes and cardio-vascular pathologies. Our study aims at studying the early impact of an obesogenic diet on the adaptive
mechanisms to fasting and re-feeding according to sex and fat depots.
We studied adipocyte and endothelial cell response to a fasting (24h) / refeeding (24h) challenge in C57Bl6/J males and females mice fed a standard (CD) or high fat (HFD, 60% kcal from fat) diet for 1 month. Stroma vascular cells and mature adipocytes from subcutaneous (AT-SC) and perigonadal (AT-PG) fat depots were isolated through enzymatic digestion. Mature adipocytes diameter was measured with a newly developed algorithm. Endothelial cells (CD45neg, CD31pos, SCA1pos) capacity to uptake fluorescent long-chain fatty acids was assessed by flow cytometry.
One month of HFD was sufficient to increase body weight in both males and females although weight gain was higher in males than in females. Interestingly we observed that fasting-induced weight loss detected in CD-fed mice was lost in males under hyperlipidic diet. This observation coincided with an alteration of lipid mobilization from adipocytes in the AT-PG of HFD-fed males. In addition, we found that early lipogenesis following refeeding observed in the AT-SC of CD-fed males was shifted into the AT-PG in HFDfed animals. In parallel we observed that AT endothelial cells (CD45neg, CD31pos, SCA1pos) ability to adapt
the rate of fluorescent fatty acids uptake according to metabolic status is impaired in HFD-fed males ATSC.
To conclude, our data show an early alteration of the regulation of fatty acid fluxes by endothelial cells and mature adipocytes in response to a fasting/refeeding under obesogenic condition. Interestingly, these defect are specific to sex and adipose depots and may contribute to the progression of metabolic disorders.
This work is supported by funds from the French National Institute for Health and Medical Research (Inserm), Université Paul Sabatier Toulouse and Région Occitanie, Association Française d’étude et de Recherche sur l’obésité (AFERO) and Société Francophone du diabète (SFD).
Metabolic adaptations in insulin resistant and dyslipidemic minipigs ranging from lean to morbid obesity
Imene Bousahba1,2*, Jérémie David1, Yves Guerin1, Benoit Cohade1, Didier Rémond1, Nathalie Poupin2 and Sergio Polakof1
1 Université Clermont-Auvergne,INRAE,UMR1019,Unité Nutrition Humaine,Clermont-Ferrand, France.
2 UMR1331 Toxalim,Université de Toulouse,INRAE,INP-Purpan,Toulouse,France
Type 2 diabetes (T2D) is a silent and slowly progressive metabolic disease to which obesity is a major contributor and whose prevention could be significantly improved if individuals at risk could be identified earlier (1). Our aim is therefore to identify phenotypes that anticipate the onset of T2D in minipigs(2) for the determination of early metabolomics-based biomarkers of T2D.
We fed four groups of minipigs (n=5-10) either on normal-fat or on a high-fat high-sugar (HFHS) diet during 2, 4 or 6 months. Body weight (BW), lean mass, fat mass (FM) and morphometric features were recorded. Biochemical, hormonal and inflammatory parameters were assessed on fasting plasma samples.
Based on multivariate statistical analysis, we observed that 4 different groups were differentiated, with 30% of the variance explained by both morphological (BW, FM) and clinical (total and HDL cholesterol, insulin, HOMA-IR and incretins) components, allowing to identify 4 distinct phenotypes: lean (LP), overweight (OWP), obese (ObP) and morbidly obese (MObP). Compared to the LP, we observed a BW increase of 2, 2.5, and 3-fold in the OWP, ObP and MobP respectively, mostly due to the FM accretion (4, 7 and 11-fold respectively). Concerning the clinical phenotype, insulin-resistance (IR) was already present in the OWP, which was further accompanied by a significant hyperinsulinemia (p = 0.03) that increases by 3.5-fold for the OWP, ObP, and MobP. Interestingly, IR affected rather lipid (2-fold increased free fatty acids) than glucose (steady glucose levels) metabolism. We also observed mild dyslipidemia with increased total cholesterol (1.3-fold) and HDL (2-fold) for all 3 phenotypes, as well as the incretin hormones (GLP-1, GIP) which actions are jointly responsible for 60% of insulin-secretion (GLP-1 p=0.02), preventing hyperglycemia. These variations are surprisingly maintained in the ObP and MObP despites of the strong weight and FM gain. The clinical phenotype was further completed by a rather pro-inflammatory pattern (slightly higher TNF-a, TGF-B) offset by a significant increase (3.7-fold) of IL-4 in the ObP, yet not confirmed in the MoBP.
Our results show that in minipigs ranging from lean to morbid obesity, IR was rapidly installed and accompanied by dysregulated lipid metabolism, while hyperglycemia was prevented by improved incretin adjustments. This ability to regulate the metabolism in the presence of progressive concomitant BW gain and adiposity with a background of IR and mild dyslipidemia makes of this model an interesting tool for the ongoing early T2D metabolomics-based biomarker’s discovery.
References
1. Chatterjee et al, The Lancet, 2017.
2. Koopmans et al, European Journal of Pharmacology, 2015.
Obesity-Hypoventilation Syndrome prevalence in patients with metabolic syndrome :
intermediate analysis estimates obesity-related sleep hypoventilation prevalence at 6%.
Mathieu Despeaux1, Rosita Lorblanches2, Nicolas Molinari3, Frédéric Sanguignol1, Delphine Delample4
1 Obesity Lab, Clinique du Château de Vernhes, F-31340 Bondigoux, France
2 Obesity Ward, Clinique d’Ursuya, F-64250 Cambo-les-Bains, France
3 Department of Medical Information, CHU Montpelier, F-34295 Montpelier, France
4 R&D Department, SOS Oxygène, F-31000, Toulouse, France
Obesity-hypoventilation syndrome (OSH) is defined as a combination of obesity (Body-Mass Index (BMI) >30kg.m-2) and hypercanip type II respiratory failure (PaCO2 ≥45mmHg) without alternative neuromuscular, mechanical, respiratory or metabolic cause of hypoventilation. Worldwide OSH prevalence is estimated to be 10-20% in obese patients with obstructive sleep apnea and 0.4% in the general adult population but is unknown in Europe. Although frequently associated with obstructive sleep apnea (OSA), it is a distinct clinical entity. The European Pneumology Society stages OSH severity from 0 (no OSH), 1-2 (obesity-related sleep hypoventilation) to 3-4 (daytime hypercapnia). Under-diagnosed, OSH is most often discovered during an acute respiratory failure, which increases health-related costs and the risk of
hospitalisation and death. It is thus critical to determine the prevalence of early stages OSH (1-2) and the clinical and environmental predictive factors within the European obese population.
Prospective multi centric observational study. Inclusion criteria were adult patients with obesity without past or current non-invasive ventilation or continus positive pressure treatment and no recent hospitalization. In this intermediate analysis, OSH prevalence, staging, BMI and associated factors (gender, age, mean peripheral saturation, night-time desaturation <90% (min
and %), arterial blood gases values, SF-36 quality of life measurement, Ricci & Ganon physical activity assessment, Epworth) were examined from the first 100 patients.
67 women and 33 men were included. Among them 6% had early stage OSH that is not routinely assessed (68% were OSA and 17% stage 3-4 OSH). Early stage OSH was associated with elevated level and duration of night-time hypercapnia (PtCO2 = 51.62mmHg ±6.88, P<0.001; PtCO2 > 50 mmHg = 144min ±166, P<0.001), hypoxemia (PaO2 = 82.55mmHg ± 14.32, P=0.015), lower pH (7.4 ± 0.03, P=0.0086) and elevated bicarbonate (HCO3- = 26.05mmol/L ± 0.92, P<0.001). No difference on BMI, sleep apnea, physical activity or Epworth score was found among groups. These results suggest that the prevalence of early stage may be of clinical relevance and independent from BMI.
This study is funded by SOS Oxygène (ID-RCB : 2019-A022866-51 project), F-34160 Castries, France.
References
Berry RB, Budhiraja R, Gottlieb DJ, et al. J Clin Sleep Med. 2012 Oct 15;8(5):597-619.
Nowbar S, Burkart KM, Gonzales R, et al. Am J Med. 2004 Jan 1;116(1):1-7.
MicroRNAs from foods resist cooking and are detected in human feces but not in serum
Ester Díez-Sainz 1*, Silvia Lorente-Cebrián 2,3,4, Paula Aranaz 1,5, José I Riezu-Boj 1,5 and Fermín I Milagro 1,5,6
1 Department of Nutrition, Food Science and Physiology/Center for Nutrition Research, Faculty of Pharmacy and Nutrition, University of Navarra, Pamplona, Spain.
2 Department of Pharmacology, Physiology and Legal and Forensic Medicine, Faculty of Health and Sport Science, University of Zaragoza, Zaragoza, Spain.
3 Instituto Agroalimentario de Aragón-IA2, Universidad de Zaragoza-CITA, Zaragoza, Spain.
4 Aragón Health Research Institute (IIS-Aragon), Zaragoza, Spain.
5 Navarra Institute for Health Research (IdiSNA), Pamplona, Spain.
6 Centro de Investigación Biomédica en Red Fisiopatología de la Obesidad y Nutrición (CIBERobn), Instituto de Salud Carlos III, 28029 Madrid, Spain
MicroRNAs (miRNAs) are gaining increasing attention due to their role in cross-kingdom gene expression regulation [1]. It has been suggested that dietary miRNAs could modulate both gut microbiota and host gene expression, influencing their physiology [2]. However, several studies refute this controversial hypothesis, suggesting that exogenous miRNAs are degraded during digestion and they will not display any biological function on the host [2,3].
The objectives of the present work are (1) to identify miRNAs from a wide range of raw and cooked plant-derived food products and (2) to determine if plant miRNAs are detected in human feces and serum after an acute ingestion of these food products. Total RNA from cooked vegetables, legumes and rice, fruits and nuts were isolated by different optimized extraction protocols, and the isolation efficiency was evaluated through the detection of conserved miRNAs by qRT-PCR. Total RNA from human feces and serum were isolated before and after an acute ingestion of plant-derived products, and human microRNAs (controls) and specific plant-derived microRNAs were identified by qRT-PCR.
The results suggest that plant-derived miRNAs (that are abundantly present in cooked vegetables, legumes, cereals, fruits and nuts) are detected in human feces samples, whose amount increase after an acute ingestion, but not in serum samples.
It is possible to isolate and quantify plant-derived miRNAs, which resist degradation during cooking. In addition, these miRNAs will not be completely degraded during digestion, being detected in feces samples. However, the unsuccessful detection of exogenous miRNAs in serum samples support the results of other studies, which suggest that dietary miRNAs will not be absorbed and will not act on a systemic level [2,3]. However, due to the fact that dietary miRNAs reach the gastrointestinal tract, they could potentially have effects at the intestinal level on gut microbiota and/or host gut resident cells.
This project is supported by CIBERobn (CB12/03/30002), MICINN (RTI2018-102205-B-I00 project) and a Center for Nutrition Research (UNAV) predoctoral grant.
References
1. Li, D.; Yang, J.; Yang, Y.; Liu, J.; Li, H.; Li, R.; Cao, C.; Shi, L.; Wu, W.; He, K. A Timely Review of Cross-Kingdom Regulation of Plant-Derived MicroRNAs. Front. Genet. 2021, 12, 613197, doi:10.3389/fgene.2021.613197.
2. Díez-Sainz, E.; Lorente-Cebrián, S.; Aranaz, P.; Riezu-Boj, J.I.; Martínez, J.A.; Milagro, F.I. Potential Mechanisms Linking Food-Derived MicroRNAs, Gut Microbiota and Intestinal Barrier Functions in the Context of Nutrition and Human Health. Front. Nutr. 2021, 8, 586564, doi:10.3389/fnut.2021.586564.
3. Micó, V.; Martín, R.; Lasunción, M.A.; Ordovás, J.M.; Daimiel, L. Unsuccessful Detection of Plant MicroRNAs in Beer, Extra Virgin Olive Oil and Human Plasma After an Acute Ingestion of Extra Virgin Olive Oil. Plant Foods Hum. Nutr. 2016, 71, 102–108, doi:10.1007/s11130-016-0534-9.
Differential remodeling of subcutaneous white and interscapular brown adipose tissue by long-term exercise training in aged obese female mice
Elisa Félix-Soriano1, Neira Sáinz1, Eva Gil-Iturbe1, Rosa Castilla-Madrigal1, Jon Celay2,3, Marta Fernández-Galilea1,4, M. Pilar Lostao1,4, José A Martínez-Climent2,3,4, María J Moreno-Aliaga1,4,5,*
1 University of Navarra; Center for Nutrition Research and Department of Nutrition, Food Science and Physiology; School of Pharmacy and Nutrition. Pamplona, Spain.
2 Division of Hematology-Oncology, CIMA, University of Navarra, Pamplona, Spain.
3 CIBERonc, Instituto de Salud Carlos III (ISCIII), Madrid, Spain.
4 IdiSNA, Navarra Institute for Health Research, Pamplona, Spain.
5 CIBERobn, Instituto de Salud Carlos III (ISCIII), Spain.
Obesity exacerbates aging-induced adipose tissue dysfunction [1,2]. This study aimed to investigate the effects of long-term exercise on inguinal white adipose tissue (iWAT) and interscapular brown adipose tissue (iBAT) of aged obese mice.
Two-month-old female mice received a high-fat diet for 4 months. Then, six-month-old diet-induced obese animals were allocated to sedentarism (DIO) or to a long-term treadmill training (DIOEX) up to 18 months of age. In exercised mice, iWAT depot revealed more adaptability, with a) increased expression of fatty acid oxidation genes (Cpt1a, Acox1), and amelioration of the inflammatory status characterized by a favorable modulation of pro/antiinflammatory genes (downregulation of Tnf and Il6, and upregulation of Adipoq and Il4) and lower macrophage (F4/80+/CD11b+) infiltration. Additionally, iWAT of trained animals showed an increment in the expression of mitochondrial biogenesis (Pgc1a, Tfam, Nrf1), thermogenesis (Ucp1), and beige adipocytes (Cd137, Tbx1) genes. In contrast, iBAT of aged obese mice was less responsive to exercise. Indeed, although an increase in functional brown adipocytes genes and proteins (Pgc1a, Prdm16 and UCP1) was observed in the DIOEX mice, few changes were found on inflammation-related and fatty acid metabolism genes. The remodeling of iWAT and iBAT depots occurred along with an improvement in the HOMA index for insulin resistance and in glucose tolerance. In conclusion, long-term exercise effectively prevented the loss of iWAT and iBAT thermogenic properties during aging and obesity. In iWAT, the long-term exercise program also reduced the inflammatory status and stimulated a fat-oxidative gene profile.
These exercise-induced adipose tissue adaptations could contribute to the beneficial effects on glucose homeostasis in aged obese mice.
Funding: MINECO-FEDER, BFU2015-65937-R), CIBERobn (CB12/03/30002), “Juan de la Cierva” grant to M. F.-G. (IJCI-2016-30025).
References
1. Reyes-Farias et al.. Biochem. Pharmacol. 2021, 192, doi:10.1016/j.bcp.2021.114723.
2. Trim et al. Front. Immunol. 2018, 9, 169, doi:10.3389/fimmu.2018.00169.
Effect of endothelial Nox5 expression in mice fed with high fat diet and in 3T3-L1 adipocytes treated with glucose and palmitic acid
Jorge G. García1,2,*, Eduardo Ansorena1,2, Fermín I. Milagro2,3, Guillermo Zalba1,2 and Carlos de Miguel1,2
1Department of Biochemistry and Genetics, University of Navarra, Pamplona, Spain.2Navarra Institute for Health Research (IdiSNA), Pamplona, Spain.3Department of Nutrition, Food Science and Physiology/Center for Nutrition Research, University of Navarra, Pamplona, Spain.
Obesity is a health issue associated with insulin resistance, altered lipid homeostasis and other pathologies [1]. One of the mechanisms involved in the development of these pathologies is the increased production of reactive oxygen species (ROS) [2]. One of the main producers of ROS is the family of nicotinamide adenine dinucleotide phosphate (NADPH) oxidases, of which NOX5 is the most recently discovered member [3]. The aim of the present work is to describe the effect of endothelial NOX5 expression on neighbouring adipose tissue in obesity conditions by using two systems.
An in-vivo model based on NOX5 conditional knock-in mice fed with a high-fat diet and an in-vitro model developed with 3T3-L1 adipocytes cultured with conditioned media of NOX5-expressing bEnd.3 endothelial cells, previously treated with glucose and palmitic acid. Animals expressing NOX5 presented lower body weight gain and reduced mesenteric and epididymal adipose mass. They also showed significantly lower glycaemia and improved insulin-induced glucose uptake accompanied by a higher expression of Glut4 and Cav1 in the adipose tissue. Accordingly, 3T3-L1 adipocytes treated with conditioned media of NOX5-expressing endothelial cells, presented similar results. Additionally, the endothelial NOX5 expression effect under obesogenic conditions seems to act through the activation of thermogenesis and lipolysis in both systems. This activation was mediated by an increase in IL-6 production derived from NOX5 activity.
As a conclusion, in response to obesogenic conditions, NOX5 endothelial activity may regulate glucose sensitivity and lipid homeostasis in the adipose tissue.
Funding: Project funded by the Ministry of Economy and Competitiveness (SAF2013-49088-R, SAF2016-79151-R). J.G.G is a fellowship from “La Caixa”.
References:
1.Bluher, M., Obesity: global epidemiology and pathogenesis. Nat Rev Endocrinol 2019, 15 (5), 288-298.
2.Fernandez-Sanchez A, Madrigal-Santillan E, et al, Inflammation, oxidative stress, and obesity. Int J Mol Sci 2011, 12 (5), 3117-32.
3.Buvelot H, Jaquet V, et al, Mammalian NADPH Oxidases. Methods Mol Biol 2019, 1982, 17-36.
Anti-steatotic effect of Gracilaria vermiculophylla extract in AML12 hepatocytes
Maitane González-Arceo1, Jenifer Trepiana1,2,3, Leixuri Aguirre1,2,3, Saioa Gómez-Zorita1,2,3, María P. Portillo1,2,3
1 Nutrition and Obesity Group, Department of Pharmacy and Food Science, Faculty of Pharmacy and Lucio Lascaray Research Center, University of the Basque Country (UPV/EHU), 01008 Vitoria-Gasteiz, Spain
2 Bioaraba Health Research Institute, 01006 Vitoria-Gasteiz, Spain
3 CIBER Physiopathology of Obesity and Nutrition (CIBERobn), Institute of Health Carlos III (ISCIII), Spain
Hepatic steatosis is defined as an excessive triglyceride accumulation in the hepatocytes [1], which is considered an early sign of metabolic disease [2]. Seaweeds are a rich source of bioactive compounds with potential health promoting properties [3]. In this context, we aimed to assess whether Gracilaria vermiculophylla extract was able to prevent triglyceride accumulation in cultured hepatocytes.
Mouse AML12 hepatocytes were incubated with palmitic acid (0.3 mM) to induce triglyceride accumulation. Simultaneously, cells were treated or not with 10, 25, 50 or 150 μg/mL G. vermiculophylla extract for 18 h. Cell viability was measured by crystal violet assay and triglyceride content was determined by spectrophotometry, using a commercial kit. Gene expression of genes involved in triglyceride metabolism was analyzed by RT-PCR at the most effective dose.
Treatment with G. vermiculophylla extract at 25, 50 and 150 μg/mL prevented triglyceride accumulation in a dose-dependent manner. Increased mRNA levels of carnitine palmitoyltransferase I (Cpt1), long chain acyl-CoA dehydrogenase (Acadl), mitochondrial transcription factor A (Tfam) and uncoupling protein 2 (Ucp2) were observed after treatment with 150 μg/mL.
G. vermiculophylla extract exerts an anti-steatotic effect, at least in part, due to an increase in fatty acid oxidation. In view of these results, an in vivo experiment will be performed to gain better understanding of the effects of G. vermiculophylla extract in hepatic steatosis and related metabolic dysfunctions.
References:
- Idilman, I.S.; Ozdeniz, I.; Karcaaltincaba, M. Hepatic Steatosis: Etiology, Patterns, and Quantification. Semin. Ultrasound CT MR. 2016, 37, 501–510.
- Bril, F. What the New Definition of Metabolic Dysfunction-Associated Fatty Liver Disease (MAFLD) Left Behind: Genetically Acquired Fatty Liver Disease (GAFLD): MAFLD heterogeneity. EBioMedicine. 2021, 72.
- Roohinejad, S.; Koubaa, M.; Barba, F.J.; Saljoughian, S.; Amid, M.; Greiner, R. Application of seaweeds to develop new food products with enhanced shelf-life, quality and health-related beneficial properties. Food Res. Int. 2017, 99, 1066-1083.
Obesity and cancer: role of adipose cells in preventing the transition from in situ to invasive breast cancer in a two-cell fluorescent spheroid model
HABANJAR Ola1, VITURET Cyrielle1, MAURIN Anne-Catherine1, DECOMBAT Caroline1, AUXENFANS Céline2, DIAB-ASSAF Mona3, CALDEFIE-CHEZET Florence1, DELORT Laetitia1
1. Université Clermont-Auvergne, INRAE, UNH, Unité de Nutrition Humaine, CRNH-Auvergne, 63000 Clermont-Ferrand;
2. Banque de tissus et cellules, Hôpital Edouard-Herriot, 69000 Lyon,
3. Equipe Tumorigénèse Pharmacologie moléculaire et anticancéreuse, Faculté des Sciences II, Université libanaise Fanar, Beyrouth, Liban
The incidence of ductal carcinoma in situ (DCIS) has increased in recent years. During the transition from DCIS to invasive ductal carcinoma (IDC), the disorganisation or even disappearance of myoepithelial cells (MECs) is observed, enhancing the ability of the cancer cells to migrate. As no marker has been defined to identify which DCIS will evolve into IDC, the understanding of this transition seems to be of major importance for better patient management. Moreover, many studies have shown the role of obesity and adipose microenvironment in breast cancer progression. In particular, our team has recently shown that adipose cells may be able to modify the functional characteristics of MECs leading to an invasion process.
Our objective is to identify preventive factors involved in the transition from DCIS to IDC in overweight or obese women by assessing the impact of the inflammatory adipose microenvironment on MECs by developing 3D models.
A fluorescent 2-cell spheroid model reproducing the pathology of DCIS is being developed. For this purpose, in situ cancer cells (MCF10DCIS-RFP) are first seeded in agarose moulds that do not allow cell adhesion. After obtaining spheroids, MECs (Hs578Bst-GFP) are added to our model, the objective being to obtain a continuous layer of MECs around the previously formed spheroids. By confocal imaging, the organisation of the cells in the spheroids as well as the rupture of the continuous MEC layer is visualised. The impact of the inflammatory adipose microenvironment will then be assessed using conditioned media derived from the culture of adipose stem cells or mature adipocytes from overweight/obese women.
For the design of fluorescent cell lines, a lentiviral transduction approach was used to obtain a stable expression of the fluorescent proteins (integrative system). MCF10DCIS cells were transduced with a RFP (Red Fluorescent Protein) gene, while MECs were transduced with a GFP (Green Fluorescent Protein) gene. These genes are under the control of constitutive promoters (EF1a or CMV type), clones of MCF10DCIS and MECs constitutively expressing GFP or RFP, respectively, which will allow to distinguish them directly under the fluorescence microscope. The 2-cell spheroid mimicking DCIS will now be reconstituted and the impact of adipose cells from women of normal-weight and obese women will be evaluated.
This project could ultimately help prevent the progression of ductal cancer in situ (DCIS) to invasive ductal cancer (IDC) and lead to the development of new therapeutic strategies for the management of overweight women with DCIS at high or low risk of developing IDC.
Identification of novel targets in adipose tissue involved in non-alcoholic fatty liver disease progression
Marta López-Yus1, María Pilar García-Sobreviela1, Raquel del Moral-Bergos1, Silvia Lorente-Cebrián1,3, José M Arbonés-Mainar1,2
1Adipocyte and Fat Biology Laboratory (AdipoFat), Translational Research Unit, University Hospital Miguel Servet, Instituto Aragones de Ciencias de La Salud (IACS).
2CIBER Fisiopatología Obesidad Y Nutrición (CIBERObn), Instituto Salud Carlos III.
3Departamento de Farmacología, Fisiología y Medicina Legal Y Forense, Universidad de Zaragoza, Instituto Agroalimentario de Aragón (IA2) (Universidad de Zaragoza-CITA), Zaragoza, Spain.
Obesity is a major risk factor for the development of Nonalcoholic fatty liver disease (NAFLD), as dysfunction of subcutaneous adipose tissue (scWAT) leads to fat accumulation in other organs. It is therefore essential to identify the molecular mechanisms underlying progression of the disease and the role played by the scWAT.
1) To identify new dysregulated genes in scWAT potentially involved in the development of NAFLD; 2) to establish an in vitro model of hMSC-derived adipocytes (hMSC) that could reflect the different stages of NAFLD; 3) to validate the expression of selected genes in our in vitro model.
Biopsies from the abdominal scWAT from 45 patients were obtained and their respective transcriptomes sequenced (RNAseq). Using a computational analysis, we obtained a set of genes upregulated in patients with elevated fatty liver index (FLI). Liver biopsies from obese patients of a different cohort were histologically analyzed and patients were grouped by NAFLD Activity Score (NAS). Paired scWAT biopsies were also obtained and hMSC were isolated and differentiated into adipocytes. Expression of selected genes was measured by qPCR in hMSC-derived adipocytes.
We observed impaired adipogenesis in hMSC-derived adipocytes as liver steatosis increases. Four genes showed a differential regulation pattern in scWAT and hMSC-derived adipocytes, increasing their mRNA expression accordingly to the degree of hepatic steatosis: SOCS3, DUSP1, SIK1 and GADD45B.
Our results constitute a proof-of-concept of the great importance of adipose-liver crosstalk and suggest that impaired adipogenic capacity of hMSC is a critical event in promoting the development of NAFLD. We propose four genes in adipose tissue as key players in NAFLD progression
Dietary proteins modulate high density lipoprotein characteristics in a sex-specific way in Apoe-deficient mice
Martínez-Beamonte, R.1, Sánchez-Marco, J. 2, Gómez-Castillo, M. 2, Herrero-Continente, T. 2, Serrano-Megías, M. 3, Botaya, D. 3, Arnal, C. 4, Barranquero, C.1, Surra, J.C. 5, Osada, J. 2, Navarro, MA. 2
1CIBER de Fisiopatología de la Obesidad y la Nutrición (CIBERobn), Instituto de Salud Carlos III, Spain.
2Biochemistry, 3Aldelis, 4Animal Pathology and 5Animal Pathology Units, Veterinary Faculty, Zaragoza University, Miguel Servet, 177, 50013 Zaragoza, Spain.
The type and amount of dietary protein has become a topic of renewed interest in light of their involvement in metabolic diseases, atherosclerosis and thrombosis. However, little attention has been devoted to the effect of avian proteins despite their wide human consumption. In a study carried out by our group we saw that compared to soybean protein, the consumption of avian proteins depending on sex resulted in similar or lower atherosclerosis development in females, and in males with better antioxidants lipoproteins, in both cases with a higher paraoxonase 1 activity. This suggests that under these conditions, the HDL lipoproteins produced a protective effect in atherosclerosis. To test this hypothesis we investigate the influence of chicken and turkey as sources of protein compared with that of soybean and the effects on HDL functionality and reverse cholesterol transport.
To this purpose, male and female Apoe-deficient were fed purified Western diets differing on their protein sources for 12 weeks. The differences between the diets, was mainly in the amino acid composition, with lower amount of soybean diet in sulfured and branched chain amino acids (BCAA). After this period, blood and liver samples were taken for analyses of HDL composition and hepatic expression of genes related to HDL metabolism (Lcat, Abca1, Scarb1, Pltp, Pon1). No significant differences were observed in APOA1 levels and the lipoprotein profiles of cholesterol, phosphatidylcholine and sphingomyelin depending on the diet protein for both sexes. Females consuming the turkey protein-containing diet showed decreased atherosclerotic foci, as evidenced by the en face atherosclerosis analyses that can be due to a higher paraoxonase 1 (PON1) activity, mainly in females. Other possible mechanism was the difference observed in the proportion of esterified and non-esterified cholesterol. The latter were associated with changes in hepatic Lcat expression, particularly In males. The intake of chicken protein caused higher levels of APOA4 compared to the others protein sources and a reduction in aortic lesion area in the turkey group vs the soybean group. Furthermore, VLDL, LDL and HDL ROS were higher in soybean group vs chicken and turkey groups accompanied with a reduction in paraoxonase activity in soybean group vs both avian groups.
Compared to soybean protein, the consumption of turkey proteins resulted in lower atherosclerosis development in females associated with changes in paraoxonase 1 activity and Lcat hepatic expression reinforcing the importance of HDL and its antioxidant properties in atherosclerosis.
Antidiabetic properties of polyphenols from Tagetes erecta and its effects on Caenorhabditis elegans fat storages
Sonia Núñez a*, Cristina Moliner a, Marta Sofía Valero b,c, Carlota Gómez-Rincón a,c, Víctor López a,c
a Department of Pharmacy, Faculty of Health Sciences, Universidad San Jorge, Villanueva de Gállego, Zaragoza. b Department of Pharmacology and Physiology, Universidad de Zaragoza. c Instituto Agroalimentario de Aragón, IA2, Universidad de Zaragoza-CITA.
Diabetes mellitus (DM) is a metabolic disease characterized by high blood sugar level that can cause severe complications to the organism or even death when no treated. However, certain dietary habits and foods may have beneficial effects for this condition. Caenorhabditis elegans is a model organism widely used for the evaluation of functional foods, nutraceuticals and bioactive compounds because approximately 60-80% of the human genes have their homologue in the C. elegans genome[1].
The polyphenols (mainly digallic acid, myricetin and laricitrin) of Tageres erecta L. were extracted from yellow and orange flowers using an ethanolic Soxhlet extraction [2]. These samples were screened for antidiabetic and anti-obesity properties using in vitro and in vivo procedures.The potential of this polyphenolic matrix was firstly tested in terms of its capacity to inhibit the enzymes lipase and α-glucosidase, as well as the inhibition of advance glycation end-products (AGEs) [3]. An obese C. elegans model was created exposing the nematode to an excess of glucose in the medium using two strains (wild-type N2 and BX24 mutant).Their fat content was quantified and compared with the effects of orlistat as reference drug using nile red staining and fluorimetry [4].
Polyphenols from both yellow and orange Tagetes erecta presented in vitro inhibitory activity against the enzymes lipase and α-glucosidase, showing lower IC50 values than acarbose. They also showed important activity preventing the glycation of proteins and reducing AGEs formation. The polyphenolic matrices reduced the fat content of obese worms in the wild-type strain (N2) with no significant differences found between negative control (100% reduction) and both tested samples (p<0.05). Meanwhile in the BX24 mutant strain, the fat reduction was considerably lower, less than 30%.
To conclude, polyphenols from Tagetes erecta can be considered as candidate bioactive compounds in the prevention and improvement of metabolic chronic diseases such as obesity and diabetes.
References
1. Shen P, Yue Y, Park Y (2018) A living model for obesity and aging research: Caenorhabditis elegans. Crit Rev Food Sci Nutr 58:741–754. https://doi.org/10.1080/10408398.2016.1220914
2. Moliner C, Barros L, Dias MI, et al (2018) Edible flowers of tagetes erecta l. As functional ingredients: Phenolic composition, antioxidant and protective effects on caenorhabditis elegans. Nutrients 10:. https://doi.org/10.3390/nu10122002
3. V. Spínola PCC (2017) Evaluation of Asteraceae herbal extracts in the management of diabetes and obesity . Contribution of caffeoylquinic acids in the inhibitory activities of digestive enzymes and formation of advanced glycation end-products (in vitro). Phytochemistry. https://doi.org/10.1016/j.phytochem.2017.07.006
4. Escorcia W, Ruter DL, Nhan J, Curran SP (2018) Quantification of lipid abundance and evaluation of lipid distribution in Caenorhabditis elegans by nile red and oil red o staining. J Vis Exp 2018:1–6. https://doi.org/10.3791/57352
Eccentric exercise and adipose tissue adaptations
Julianne TOURON1, Jean-Paul RIGAUDIERE1, Christophe MONTAURIER1, Ruddy RICHARD1,2, Frédéric CAPEL1.
1UCA-INRAE UMR1019, France, 2University hospital of Clermont-Ferrand, France.
Disturbances in adipose tissue (AT) metabolism related to inflammation regulation and adipokines production lead to alterations in its capacity to store lipids and release fatty acids (FA) during feeding/fasting transition or when energy demand increases. Exercise is an optimal way to maintain AT health but conventional trainings are not always suitable for patients with functional limitations. Dynamic eccentric (ECC) exercise can overcome those limitations and prevent the accumulation of AT. Consequently, we studied if ECC could improve AT health and if changes in AT metabolism could contribute to the beneficial adaptations observed at the whole body level.
60 rats were randomly assigned to a control (CTRL) group or in one of the three-trained groups, where treadmill slopes modulate exercise oxygen consumption (VO2) and mechanical work: 1) +15% uphill-concentric group (CON, ~60% of VO2max), 2) -15% downhill group (ECC15, same work as CON) and 3) -30% downhill group (ECC30, same VO2 as CON). Body composition and energy expenditure (EE) were measured before and after the 8-week training intervention. After sacrifice, abdominal subcutaneous AT was collected to study total FA profile by GCFID and gene expression by RT-qPCR.
No fat mass gain was observed for trained animals although they exhibited a significantly increased total body mass. Furthermore, their higher total EE was driven by their lean mass gain. In AT, there was a decrease in the amounts of palmitic, palmitoleic and arachidonic acids, whereas there was a trend of an increase in AT stearic acid in trained groups. Exercise also tended to decrease AT apelin expression in all group while GLUT4 remain unchanged. Increased adiponectine and IGF-1 mRNA levels were observed in ECC15 group compared to CTRL. No signs of browning (PRDM16) or change in inflammatory status (TNFα) were observed.
ECC training changes body composition whether at low or higher exercise metabolic requirements. Exercise is also in favor of modification of AT fatty acid profile and expression of key genes related to metabolism, insulin sensitivity, suggesting that ECC is a relevant for the management of overweight and obesity.
Julianne TOURON1, Jean-Paul RIGAUDIERE1, Christophe MONTAURIER1, Ruddy RICHARD1,2, Frédéric CAPEL1.
1UCA-INRAE UMR1019, France, 2University hospital of Clermont-Ferrand, France.
Disturbances in adipose tissue (AT) metabolism related to inflammation regulation and adipokines production lead to alterations in its capacity to store lipids and release fatty acids (FA) during feeding/fasting transition or when energy demand increases. Exercise is an optimal way to maintain AT health but conventional trainings are not always suitable for patients with functional limitations. Dynamic eccentric (ECC) exercise can overcome those limitations and prevent the accumulation of AT. Consequently, we studied if ECC could improve AT health and if changes in AT metabolism could contribute to the beneficial adaptations observed at the whole body level.
60 rats were randomly assigned to a control (CTRL) group or in one of the three-trained groups, where treadmill slopes modulate exercise oxygen consumption (VO2) and mechanical work: 1) +15% uphill-concentric group (CON, ~60% of VO2max), 2) -15% downhill group (ECC15, same work as CON) and 3) -30% downhill group (ECC30, same VO2 as CON). Body composition and energy expenditure (EE) were measured before and after the 8-week training intervention. After sacrifice, abdominal subcutaneous AT was collected to study total FA profile by GCFID and gene expression by RT-qPCR.
No fat mass gain was observed for trained animals although they exhibited a significantly increased total body mass. Furthermore, their higher total EE was driven by their lean mass gain. In AT, there was a decrease in the amounts of palmitic, palmitoleic and arachidonic acids, whereas there was a trend of an increase in AT stearic acid in trained groups. Exercise also tended to decrease AT apelin expression in all group while GLUT4 remain unchanged. Increased adiponectine and IGF-1 mRNA levels were observed in ECC15 group compared to CTRL. No signs of browning (PRDM16) or change in inflammatory status (TNFα) were observed.
ECC training changes body composition whether at low or higher exercise metabolic requirements. Exercise is also in favor of modification of AT fatty acid profile and expression of key genes related to metabolism, insulin sensitivity, suggesting that ECC is a relevant for the management of overweight and obesity.
Pediococcus acidilactici CECT9879 alleviates metabolic syndrome and obese-related dysbiosis in rats
Deyan Yavorov1 2; Fermín I. Milagro1 3 4; Josune Ayo2; María Oneca2; Paula Aranaz1 3
1 Center for Nutrition Research, University of Navarra, Pamplona, Spain
2 Genbioma Aplicaciones SL. Polígono Industrial Noain-Esquiroz, Calle S, Nave 4, Esquíroz, Navarra, Spain
3 Navarra Institute for Health Research (IdiSNA), Pamplona, Spain
4 Centro de Investigación Biomédica en Red de la Fisiopatología de la Obesidad y Nutrición (CIBERObn), Instituto de Salud Carlos III, Madrid, Spain
Different probiotics and bioactive compounds have emerged as a natural alternative to the classic treatments of metabolic syndrome-related diseases such as obesity, insulin resistance and diabetes. Furthermore, it has been demonstrated the importance of gut microbiota in the development of the host immunity and in the regulation of inflammation and energy homeostasis.
1) To evaluate the effects of the supplementation with the probiotic Pediococcus acidilactici CECT9879 (PA) on the regulation of adiposity and lipid metabolism in diet-induced obese rats. 2) To study the effect of PA supplementation on gut microbiota composition.
To induce obesity, 4-week-old male Wistar rats were fed a high fat/high sucrose diet (HFHS) for 4 weeks. One subgroup (n=10) was supplemented with the probiotic during 9 weeks and compared with placebo (n=11). Fat depots, liver and other organs were weighed and different serum biochemical parameters were analyzed. Gene expression analyses in adipose tissue were conducted by real-time quantitative-PCR. Fecal microbiota composition was evaluated by 16s metagenomics.
Although no effects were observed on body weight, animals supplemented with PA exhibited lower proportion of visceral adiposity. A downregulation of the circulating levels of two inflammatory molecules was found: oxidized low-density lipoprotein (ox-LDL) and monocyte chemotactic protein-1 (MCP-1). An improvement in the total-Cholesterol/HDL-Cholesterol ratio was also observed. Gene expression analyses showed an upregulation of key genes of the adipose metabolism (Adipoq, Pparg, Srebf) and glucose metabolism (Slc2a1, Slc2a4). The metagenomics study demonstrated an increase in the alpha diversity (Simpson diversity index, p=0.036) and the presence of PA in the fecal samples (this microorganism being not present in the non-treated group).
Our data suggest that PA could be a potential probiotic strain for the prevention of the metabolic syndrome-related disturbances and may help to fight obesity-related dysbiosis.