9th Congress of the
International Society of
Nutrigenetics/Nutrigenomics
(ISNN)
May 17–19, 2015
Chapel Hill, N.C., USA
Guest Editor
Martin Kohlmeier, Chapel Hill,
N.C., USA
Abstracts
J Nutrigenet Nutrigenomics 2015;8:1–25 DOI: 10.1159/000430793
Published online: May 12, 2015
Basel · Freiburg · Paris · London · New York · Chennai · New Delhi · Bangkok · Beijing · Shanghai · Tokyo · Kuala Lumpur · Singapore · Sydney
Abstracts
1
Vitamin D Receptor Polymorphism Fok1
Alters Risk of Adenomatous Polyps in
Australian Males
E.L. Beckett1,2, K.C. le Gras1, M. Veysey3, L. Boyd1, X. Ng1, Z. Yates4, K. Duesing2, M. Lucock1
1School of Environmental and Life Sciences, University of Newcastle, Central Coast, Ourimbah, NSW, Australia; 2Food and Nutrition Flagship, CSIRO, North Ryde, NSW, Australia; 3Central Coast Local Health District, Gosford, NSW, Australia; 4School of Biomedical Sciences and Pharmacy, University of Newcastle, Central Coast, Ourimbah, NSW, Australia
E-mail: [email protected]
Background/Aims: The vitamin D receptor (VDR) gene poly-morphism, Fok1 (rs10735810), may influence risk of cancers via modulation of multiple vitamin D sensitive pathways, including angiogenesis, cell proliferation and differentiation, and protection from oxidative stress. In colorectal cancer (CRC) however, results have been mixed and any association remains contentious [1–7]. Failure to clinically exclude the presence of adenomatous polyps (AP), the benign precursor of CRC, in control cohorts may contribute to the lack of consensus. Insufficient adjustment for other potential risk factors (such as diet, smoking and alcohol intake) may also explain some of the discrepancies. Therefore, we assessed the role of the Fok1 polymorphism in modifying the risk for AP in clinically confirmed cases and controls, adjusting for a range of dietary and lifestyle variables.
Methods: Blood was collected from patients undergoing routine colonoscopy (n = 258). Diagnosis of AP was used to classify cases and controls. Fok1 polymorphisms were assessed using RFLP-PCR (F = absence; f = presence of restriction site, early start codon, less transcriptionally active) [8–9]. Dietary habits were estimated from food frequency questionnaires. AP incidence (OR, 95% CI) were cal-culated by genotype, stratified by sex and corrected for age. Additional adjustments were made for life-style factors (smoking his-tory and alcohol intake), objective markers of diet (blood B12 and folate), and reported markers of dietary habits (including estimated dietary fibre, iron, vitamin C, calcium, vitamin D and fat intake). Regression analysis was used to examine the relationship between vitamin D and AP risk.
Results: Participants were aged 18–89 (mean 62.25 ± 0.75); 44% were male. AP were detected in 57 (22%) of participants. No statistically significant relationships were found between AP and
Fok1 genotype in females. In males the ‘F’ allele was associated with increased incidence of AP. When adjusted for age only, AP incidence was increased in ‘F’ homozygotes, relative to ‘f’ homozygotes (OR = 6.43, 1.09–123.03). Additional adjustment for lifestyle factors revealed increased AP incidence in those possessing the ‘F’ allele (ORs: FF = 7.59, 1.23–148.70; Ff = 6.42, 1.07–124.68). Further adjustment for dietary variables revealed similar associations. Reported vitamin D intake did not correlate with risk of AP in males or females, regardless of correction for age, smoking status, alcohol consumption or dietary factors (p > 0.05).
Conclusion: In Australian males, VDR-Fok1 polymorphisms may influence the risk of AP, the benign precursor to CRC, which in turn, is likely to influence the risk of CRC. Lifestyle & dietary habits influence the strength of this association and need to be fully consid-ered in future studies. This study offers novel insight into the poten-tial for VDR genetics to contribute to risk for adenomatous polyps, and is the first to demonstrate a sex-specific relationship between the VDR-Fok1 polymorphism and risk for adenomatous polyps.
References
1 Ingles, et al: Cancer Causes Control 2001;12:607–614.
2 Jenab, et al: Cancer Epidemiol Biomarkers Prev 2009;18:2485–2491. 3 Mahmoudi, et al: Mol Biol Rep 2011;38:4765–4770.
4 Murtaugh, et al: Nutr Cancer 2006;55:35–43. 5 Park, et al: Cancer Lett 2006;237:199–206.
6 Peters, et al: Cancer Epidemiol Biomarkers Prev 2001;10:1267–1274. 7 Wong, et al: Carcinogenesis 2003;24:1091–1095.
8 Whitfield, et al: Mol Cell Endocrinol 2001;177:145–159. 9 Jurutka, et al: Rev Endocr Metab Disord 2001;2:203–216.
2
Heart Rate Variability and Cardiovascular
Risk Reduction in Women after Bariatric
Surgery
M. Souza Pinhel1, Bruno Affonso Parenti Oliveira1, Camila Balsamo Gardim1, Carolina Ferreira Nicoletti1, Driele Cristina Gomes Quinhoneiro1,
Cristiana Cortes-Oliveira1, Dorotéia Silva Souza2, Wilson Salgado Junior1, Moacir Fernandes Godoy2, Carla Barbosa Nonino1
1Faculty of Medicine of Ribeirao Preto, University of Sao Paulo, FMRP/USP, Ribeirao Preto, Sao Paulo, Brazil; 2Sao Jose do Rio Preto Medical School, FAMERP, Sao Jose do Rio Preto, São Paulo, Brazil
E-mail: [email protected]
Objective: To evaluate linear and nonlinear HRV indexes in women with morbid obesity before and after bariatric surgery for Roux-Y gastric bypass (RYGB) technique.
Methods: Weight (kg), body mass index (BMI-kg/m2) and HRV were evaluated in 13 women with morbid obesity before and after the postoperative period of 6 months. Time series were collected from 1000 RR intervals extracted from electrocardiograms obtained for 30 minutes. Statistical analysis Results were presented as mean ± stan-dard deviation and compared by paired t test or the test Mann-Whitney test (SPSS 17.0; p < 0.05).
Results: The mean age of patients was 38.0 ± 11.0 years. The average weight decreased from 116.2 ± 91.0 ± 16.4 kg to 13.4 kg; p < 0.0001. There were very low HRV values preoperatively rising sig-nificantly in the evaluation at 6 months after surgery (SDNN 49.8 ± 28.5 ms versus 70.3 ± 33.1 ms; p = 0.009; RMSSD 44.8 ± 34.7 ms versus 72.7 ± 38.9 ms; p = 0.006; pNN50 211.0 ± 418.0 ms versus 224.7 ± 220.7 ms; p = 0.004; LF 756.4 ± 1049.6 ms2 versus 1486.7 ± 1417.2 ms2; p = 0.022; SD1 31.7 ± 51.4 ms2 versus 24.5 ms2 ± 27.5 ms2, p = 0.006; SD2 62.5 ± 32.9 ms2 versus 84.2 ± 39.9 ms2, p = 0.018). The non-linear indices indicated recovery of complex behav-ior, away from the more linear behavior significantly (ShanEnt 3.07 ± 0.33 versus 2.83 ± 0.27 p = 0.006; Lmean 10.26 ± 2.87 versus 8.16 ± 1.92 p = 0.006).
Conclusion: This study shows improvement in linear fields and nonlinear HRV in women with morbid obesity after 6 months of bar-iatric surgery, indicating that significant weight loss reflects homeo-stasis via the autonomic nervous system with increased HRV suggesting that this may be a mechanism for reducing the risk of car-diovascular disease. The next step we will associate these results with whole transcriptome to identify molecular pathways involved in chronic degenerative diseases.
3
A Mediterranean-Style Breakfast Can
Induce Short-Term Improvements in
Circulating Inflammatory Markers and
in Pro-Inflammatory Genes
Milena Monfort-Pires, R. Amanda Crisma, A. Silvana Bordin, R.G. Sandra Ferreira University of São Paulo, São Paulo, Brazil E-mail: [email protected]
Objective: There is evidence that association of certain dietary patterns and cardiometabolic profile are mediated by inflammation. This study investigates the effects of 2 isocaloric breakfasts, a typical Brazilian (BrB) and a ‘mediterranized’ breakfast (MedB), in cardio-vascular risk factors, circulating inflammatory markers and expres-sion of pro-inflammatory genes.
Methods: This crossover clinical trial included 80 overweight individuals with at least one additional cardiovascular risk factor. Participants received one of two breakfasts for 4 weeks. In a random order, either a Brazilian – BrB (coffee, whole fat milk, french bread, butter and mozzarella cheese) or Mediterranean breakfast – MedB (coffee, low-fat milk, whole bread, ricotta cream with olive oil and peanuts) was given to participants. After a 2-week washout, individu-als received the other intervention. Gene expression was assessed by PCR array in a pooled sample (with 5 individuals) using atherosclero-sis pathway (Qiagen®). Variables before and after each intervention were compared by Students’ t test.
Results: Participants (51.7 ± 9.5 years) had a mean BMI of 30.5 ± 4.2 kg/m2 and waist circumference of 99.7 ± 10.1 cm. Body weight did not change during interventions. After 4 weeks of inter-vention, the consumption of BrB increased (p < 0.05) the concentra-tions of TNF-α (3.32 ± 1.88 to 6.13 ± 1.90 pg/ml), IL-1β (1.61 ± 0.42 to 3.00 ± 6.9 pg/ml), IL-17 (2.09 ± 0.7 to 4.21 ± 1.22 pg/ml), INF-γ (1.52 ± 0.58 to 2.79 ± 0.77 pg/ml) and E-selectin (12.6 ± 5.0 to 17.6 ± 7.0 ng/ml). The MedB induced reductions (p < 0.05) in CRP concen-trations (0.85 ± 0.1 to 0.78 ± 0.1 mg/L) and MCP-1 (1073.3 ± 482.9 to 583.8 ± 396.9 pg/ml, p < 0.01) and also improvements in other cardiometabolic risk factors such as waist circumference (99.2 ± 10.4 to 98.5 ± 10.5 cm, p = 0.02) and HDL-c (51.8 ± 13.5 to 54.0 ± 13.9 mg/dl, p = 0.01). Reductions in other inflammatory markers, such as E-selectin (13.7 ± 5.9 to 12.9 ± 7.24 ng/ml) and INF-γ (2.01 ± 0.9 to 1.81 ± 1.0 pg/ml) were not significant, p > 0.05). PCR array of a pooled subsample of individuals showed that the MedB decreased the expression of pro-inflammatory genes such as interleukin-1, chemo-kine ligand 2 (CCL2), TNF-α, INF receptor 2, VCAM-1, colony stimulating factor-2, selectin-E and angiotensin I converting enzyme, while BrB increased the expression of the same genes.
Conclusions: Our data indicate that modification of a single meal may improve inflammatory status and cardiometabolic risk fac-tors, such as HDL-c and waist circumference. Even a short interven-tion with Mediterranean foods was able to induce changes in the expression of pro-inflammatory genes in a Brazilian sample. Confirmation of these findings of improved inflammatory profile could motivate changes in eating habits in non-Mediterranean coun-tries like Brazil.
4
Coupling Genetic Addiction Risk Score
(GARS
DX)
TMand a Nutrigenomic
Neuroadaptagen Dopaminergic Reward
Brain Activator to Treat Reward Deficiency
Syndrome (RDS): Targeting Polymorphic
Reward Genes for Addiction Algorithms
Kenneth Blum1–8, Thomas Simpatico3,6, A. Margaret Madigan5, L. Roger Waite2, C. Brett Haberstick9, Andrew Smolen9, D. Rajendra Badgaiyan10, Zsolt Demotrovics11, B. William Downs7, Gozde Agan6, James Fratantonio6, John Giordano8, Marcelo Febo1
1Department of Psychiatry & McKnight Brain Institute, University of Florida College of Medicine, Gainesville, FL, USA; 2Department of Nutrigenomics, RDSolutions Inc., Del Mar, CA, USA; 3Department of Addiction Research & Therapy, Malibu Beach Recovery Center, Malibu Beach, CA, USA; 4Department of Psychiatry, Human Integrated Services Unit, University of Vermont Center for Clinical & Translational Science, University of Vermont College of Medicine, Burlington, VT, USA; 5Department of Personalized Addiction Medicine IGENE, LLC, Austin, Texas, USA; 6Dominion Diagnostics, LLC., North Kingstown RI, USA; 7Department of Nutrigenetic & Nutrigenomic Research, Victory Nutrition International , Austin, TX, USA; 8National Institute of Holistic Addiction Studies, North Miami Beach, FL,USA; 9Institute for Behavioral Genetics, University of Colorado Boulder, Boulder, CO, USA; 10Department of Psychiatry, University of Minnesota , College of Medicine, MN, USA; 11Eotvos Lorand University, Institute of Psychology, Department of Clinical Psychology and Addiction, Izabella utca, Budapest, Hungary
E-mail: [email protected]
We are entering the era of genomic medicine and neuroimaging as it relates to addiction a subset of Reward Deficiency Syndrome (RDS) [1]. In 2005 our laboratory received the first USA patent on Nutrigenomics and RDS treatment. This was awarded on the basis of our earlier work showing anti-addiction activity of a nutraceutical consisting of amino-acid precursors and enkephalinase inhibition properties and our discovery of the first polymorphic gene (Dopamine D2 Receptor Gene [DRD2] to associate with sever alcoholism [2]. Prior to the later genetic finding we developed the concept of Brain Reward Cascade which continues to act as blue-print for stratification of addiction risk through neurogentics. In 1996 our laboratory also coined the term ‘Reward Deficiency Syndrome (RDS)’ to define a common genetic rubric for both substance and non-substance related addictive behaviors [1]. Following many reiterationswe utilized poly-morphic targets of a number of reward genes (serotonergic, Opioidergic, GABAergic and Dopaminergic) to customize KB220 [Neuroadaptogen-amino-acid therapy (NAAT)] by specific algo-rithms. Identifying 1,000 obese subjects in the Netherlands a subse-quent small subset was administered various KB220 formulae customized according to respective DNA polymorphisms individual-ized that translated to significant decreases in both Body Mass Index (BMI) and weight in pounds [3]. This was followed up in the USA
with similar significant effects. Following these experiments we have been successfully developing a panel of genes known as ‘Genetic Addiction Risk Score (GARSDX)TM. When we selected 10 genes with appropriate variants, a statistically significant association between the ASI-Media Version-alcohol and drug severity scores and GARSDx was found. This observation was found in 273 patients attending seven diverse treatment centers. Independently, a variant of NAAT– KB220Z in abstinent heroin addicts, remarkably increased resting state functional connectivity was observed in a putative network that included the dorsal anterior cingulate, medial frontal gyrus, nucleus accumbens, posterior cingulate, occipital cortical areas, and cerebel-lum [4]. In other unpublished rat work we show that KB220Z signifi-cantly activates, above placebo, seed regions of interest including the left nucleus accumbens, cingulate gyrus, anterior thalamic nuclei, hippocampus, pre-limbic and infra-limbic loci. This response induced by KB220Z demonstrates significant functional connectivity, increased brain volume recruitment and enhanced dopaminergic functionality across the brain reward circuitry. This robust yet selec-tive response implies clinical relevance. We are now paused to pro-pose a Reward Deficiency System Solution that promotes early identification and stratification of risk alleles by utilizing GARSDx allowing for customized nutrigenomic targeting of these risk alleles by altering NAAT ingredients as an algorithmic function of carrying these polymorphic DNA – SNPS potentially yielding the first ever nutrigenomic solution for addiction and pain.
References
1 Blum K, Oscar-Berman M, Stuller E, Miller D, Giordano J, Morse S, McCormick L, Downs WB, Waite RL, Barh D, Neal D, Braverman ER, Lohmann R, Borsten J, Hauser M, Han D, Liu Y, Helman M, Simpatico T: Neurogenetics and Nutrigenomics of Neuro-Nutrient Therapy for Re-ward Deficiency Syndrome (RDS): Clinical Ramifications as a Function of Molecular Neurobiological Mechanisms. J Addict Res Ther 2012; 3:139.
2 Blum K, Noble EP, Sheridan PJ, Montgomery A, Ritchie T, Jaga-deeswaran P, Nogami H, Briggs AH, Cohn JB: Allelic association of human dopamine D2 receptor gene in alcoholism. JAMA 1990;263: 2055–2060.
3 Blum K, Chen TJH, Williams L, Chen ALC, Downs BW, Waite RL, et al: A short term pilot open label study to evaluate efficacy and safety of LG839, a customized DNA directed nutraceutical in obesity: exploring nutrigenomics. Gene Ther Mol Biol 2008;12:371–382.
4 Blum K, Liu Y, Wang W, Wang Y, Zhang Y, Oscar-Berman M, Smolen A, Febo M, Han D, Simpatico T, Cronjé FJ, Demetrovics Z, Gold MS: rsfMRI effects of KB220ZTM on neural pathways in reward circuitry of
5
Protective Effect of Glutathione on Damage
Induced by Permethrin in a Neuronal Model
of PC12 Cells
Bordoni Laura1, Capitani Melania2, Nasuti Cinzia2, Gabbianelli Rosita2
1School of Advanced Studies and 2School of Pharmacy, University of Camerino, Italy
E-mail: [email protected]
Objective: Permethrin, a member of the family of synthetic pyrethroids, can induce oxidative stress and impairment in expression of Nurr1, a transcription factor essential for the maintenance of dopa-minergic neurons. These conditions, flanked to low level of glutathi-one (GSH), are typical in neurons affected by Parkinson Disease (PD). The aim of this study is to evaluate in vitro effects of GSH on a stress dependent parameters (Nurr1) measured in PC12 cells culture treated with permethrin.
Methods: PC12 cells were seeded and treated for 72 h with 1 μM
permethrin and separately co-treated with the pesticide and 32 nM
glutathione. Then, qRT-PCR was performed to evaluate Nurr1 gene expression.
Results: In cells cultures under stress condition, such as those induced by permethrin, Nurr1 gene expression changed, maybe as a compensatory effect to damage. In contrast, when cells were co-treated with glutathione, Nurr1 value was reestablished as the control. Conclusion: Pesticides, mainly assumed with diet, can modulate gene expression leading to the onset and progression of age-related diseases. Permethrin, in particular, can induce stress in neuronal dopaminergic cells, mimicking PD condition. As demonstrated by previous in vivo studies, permethrin exposure can lead to progressive neuronal damage characterized by GSH and Nurr1 deficit. The pres-ent study shows that GSH in vitro can contrast the negative effect induced by permethrin. We can hypothesize that GSH can prevent oxidative stress damage, avoiding cells to overreact by modifying the production of Nurr1. Hence, GSH can act as a preventive agent to cellular damage induced by permethrin.
6
Permethrin Pesticide Residues in Food
Mediate Progressive Neuronal Disorder
Nasuti Cinzia1, Vincenzetti Silvia2, Correia-Sá Luísa4, Domingues Valentina4, Fedeli Donatella1,
Ricciutelli Massimo3, Pucciarelli Stefania2, Gabbianelli Rosita1
1School of Pharmacy, 2School of Bioscience and
Veterinary Medicine, and 3HPLC-MS Laboratory, University of Camerino, Camerino, Italy; 4REQUIMTE/LAQV, Instituto Superior de Engenharia do Instituto Politécnico do Porto, Porto, Portugal
E-mail: [email protected]
Objective: Pyrethroids are used in many formulations in agricul-ture to control insect pests in crops, forestry and horticulagricul-ture. Environmental exposure results from dietary intake and this is con-firmed by the quantification in human urine of the 3-PBA pyrethroid metabolite. The exposure to xenobiotics in the early stages of life, represents the most important component in the etiology of neurode-generative diseases. Here, the impact of permethrin administered to rats from 6th to 21st day of life, was studied in adult animals by pro-teomic analysis of plasma and striatum. The concentration of perme-thrin and its 3-PBA metabolite were also evaluated.
Methods: GC-MS/SIM and GC-ECD were used to detect 3-PBA and permethrin in urine and striatum samples of rats 24 h and 14 days after the treatment. Two-dimensional electrophoresis followed by LC-MS/MS analysis was used for proteomic analysis.
Results: Permethrin in the brain was found at either 24 h or 14 days after the end of treatment and it was 10 times higher at 14 days; while the 3-PBA level was higher at 24 h in both brain and urine. In plasma, long chain fatty acid protein transporter resulted significantly decreased on treated animal compared to control group. In striatum, mitochondrial Aspartate aminotransferase and voltage dependent anion channel were down expressed while Malate dehydrogenase, Myelin basic protein and Ubiquitin-60S ribosomal protein L40 resulted completely absent.
Conclusion: Permethrin accumulation in striatum can be respon-sible for the long term effects reported in striatum leading to down expression of proteins associated with neuronal disorder. Proteomic data can be utilized as biomarkers of neurodegeneration.
7
Acute Intake of Curcumin Differently
Affects Gene Expression in Immune Cells
and Vascular Function in Middle-Aged
Healthy Male and Female Smokers
D. Milenkovic1, N. Barber-Chamoux1, M.A. Verny1, V. Habauzit2, C. Boby3, A. Mazur1, C. Dubray2, C. Morand1 1INRA, Unité de Nutrition Humaine, UMR 1019, Centre de Recherche de Clermont-Ferrand/Theix, 63122 Saint Genès Champanelle, France; 2Centre Investigation Clinique, CIC-CPC INSERM 501, Centre Hospitalier Universitaire, 63000 Clermont-Ferrand, France; 3INRA, Unité Mixte de
Recherche sur Herbivores, Centre de Recherche de Clermont-Ferrand/Theix, 63122 Saint Genès Champanelle, France
E-mail: [email protected]
Objective: Curcumin, principal curcuminoid in turmeric, is obtained from the rhizomes of Curcuma longa. This compound exhibits potent antioxidant, anti-inflammatory, immunomodulatory or anti-angiogenic properties. Several animal studies reported hypo-cholesterolemic, antioxidant, anti-atherogenic and antiplatelet activi-ties of this compound, however the cardiovascular protective effect of curcumin was poorly investigated in humans. The aim of this study was to determine the effect of an acute intake of curcumin on vascular endothelial function in healthy smokers and its nutrigenomic impact in circulating immune cells.
Methods: In a randomized, double-blind, cross-over study on 18 smokers (9 men and 9 women), 50–70 yrs old, the effect of curcumin (5 g) on gene expression in peripheral blood mononuclear cell (PBMC) and endothelial function was evaluated. Effect on expres-sion of 93 genes involved in inflammation, cell adheexpres-sion, cholesterol absorption/efflux was assessed using TaqMan-Low-Density-Array (TLDA) and endothelial function was evaluated by Flow Mediated Dilatation (FMD). Measurements were done before and 2-hours after curcumin or placebo intake.
Results: We failed to show significant difference between cur-cumin and placebo for FMD partly due to inter-individual variability. However, analysis of data separately for men and women revealed a significant effect of curcumin for FMD in women, but not in men. Similarly, no change in gene expression between curcumin and pla-cebo was observed when data were analysed for all volunteers. However, analysis of gene expression data separately for men and women revealed 9 differentially expressed genes in men and 8 in women with 3 genes in common (FABP3, CXCL3, CCL20). Interestingly, the expression of the 9 and 8 genes was identified as up-regulated in men and down-regulated in women, respectively.
Conclusion: Although results did not account for a beneficial effect of curcumin in the whole studied population, gender compari-son analysis suggests a difference in nutrigenomic and vascular response to curcumin between men and women.
8
Transcriptomic and Epigenomic Changes in
Human Leukocytes Upon 8 Weeks
Supplementation with Monomeric and
Oligomeric Flavanols
D. Milenkovic1, W. Vanden Berghe2,3, K. Heyninck3, K. Szarc vel Szic2, F. Fuks4, C. Gerhauser5, G. Haegeman3, G.R.M.M. Haenen6, A. Bast6, A.R. Weseler6
1INRA Research Centre Clermont-Ferrand/Theix, Clermont-Ferrand, France; 2University of Antwerp, Antwerp, Belgium; 3University of Gent, Gent, Belgium; 4Free University of Brussels, Brussels, Belgium; 5DKFZ Heidelberg, Heidelberg, Germany; 6Department of Toxicology, Maastricht University, Maastricht, The Netherlands
E-mail:[email protected]
Objective: Consumption of flavanol-rich foods is associated with a reduced risk of cardiovascular diseases, which was linked to improvements in endothelial function. A recent randomized, double-blind, placebo controlled clinical trial unveiled pleiotropic health benefits in the vasculature of healthy male smokers upon 8 weeks supplementation with daily 200 mg flavanols. In order to unravel the flavanols’ underlying molecular mechanisms, we investigated the transcriptomic and epigenomic changes in leukocytes.
Methods: Gene expression profiles were determined using whole genome microarrays (Agilent) and DNA methylation was assessed using HumanMethylation450 BeadChips (Illumina).
Results: Flavanol consumption significantly modulated the expression of 864 genes. The majority of the affected genes are involved in chemotaxis, cell adhesion, cell infiltration or cytoskeleton organisation, suggesting lower immune cell adhesion to endothelial cells. This was corroborated by in vitro experiments showing that fla-vanols exposure of monocytes attenuates their adhesion to TNF-α-stimulated endothelial cells. Nuclear factor kappa B (NF-κB) reporter gene assays confirmed that flavanols decrease the activity of NF-κB. Strong inter-individual variability in the leukocytes’ DNA methyla-tion was observed. As a consequence, on group level, changes due to flavanols supplementation could not be found.
Conclusion: Altogether, flavanols may elicit protective effects in the vasculature by decreasing inflammatory and cell adhesion pathways at the transcriptional level. Moreover, smoking history may be a confounding factor in epigenetic profiling studies of leukocytes from subjects involved in a flavanol-rich diet intervention.
9
Mammary Nutrigenomics in Lactating Goats
Leroux Christine, Boby Céline, Bernard Laurence, Chilliard Yves, Faulconnier Yannick
INRA, UMR1213 Herbivores, F-63122
Saint-Genès-Champanelle, France; Clermont Université, VetAgro Sup, UMR1213 Herbivores, BP 10448, F-63000, Clermont-Ferrand, France
E-mail: [email protected]
Objective: In ruminants, nutritional strategies were developed with dietary supplementations such as plant oils or seeds rich in n-3 polyunsaturated FA to increase the milk nutritional value. Elsewhere, milk components biosynthesis in mammary gland (MG) involves a large number of genes which nutritional regulation is not totally known. So, the objective of the present study was to evaluate the effects of the addition of extruded linseed (EL) alone or in combination with fish oil (ELFO) on MG gene expression profile in lactating goats.
Methods: Fourteen Alpine goats in mid-lactation were used to compare the effects of 3 diets: natural grassland hay control diet (CTRL) alone or supplemented with 530 g/d of EL or with 340 g/d of extruded linseed plus 39 g/d of fish oil (ELFO) on mammary tran-scriptomes. Nutrigenomics analyses were performed to compare EL or ELFO with CTRL diet using a bovine microarray which the proce-dure was previously described by Faulconnier et al. (2011).
Results: The classification of the differentially expressed genes (DEG; 344 with EL vs CTRL and 314 with ELFO vs CTRL) pointed out the cell cycle, proliferation, differentiation and death together with the protein metabolism and transport classes as the two main categories altered by EL or ELFO diets. This last class highlighted signaling pathway involving mTOR. Moreover, two networks cen-tered on an estrogen receptor (ESR1) and a transcriptional factor (SP1) both with EL and ELFO diets compared to CTRL diet were identified.
Conclusions: The comparison of gene expression using tran-scriptomic approach pointed out different networks (ESR1 and SP1) or mTOR signaling pathway which must be considered and need fur-ther investigations to precise their respective role.
10
Food Deprivation Affects the Goat
Mammary miRNome
Mobuchon Lenha1,2, Marthey Sylvain1, Le Guillou Sandrine1, Laloë Denis1, Le Provost Fabienne1, Leroux Christine2 1INRA, UMR1313 Génétique Animale et Biologie Intégrative, F-78350, Jouy-en-Josas, France; 2INRA, UMR1213 Herbivores, F-63122 Saint-Genès-Champanelle, France
E-mail: [email protected]
Objective: Nutrition affects milk composition thus influencing its nutritional properties. Nutrition also modifies the expression of mammary genes, whose regulation is not fully understood.
MicroRNAs (miRNA) are small non coding RNA which are important post-transcriptional regulators of gene expression by targeting messenger RNAs. Our goal was to characterize miRNA whose expression is regulated by nutrition in the lactating goat mammary gland (MG), which may provide clues to decipher-ing regulations of the biosynthesis and secretion of milk compo-nents.
Methods: RNA samples from MG of Alpine goats at peak lacta-tion (48 ± 2 days) were used to construct ten libraries from the MG of four Control and six food-deprived goats using the Illumina/Solexa technology. After cleaning, reads were filtered according to their size (17–28 nt). Data analyses were processed using mostly miRDeep2 software as described by Le Guillou et al. (2014). The cleaned sequences were clustered into unique reads and then mapped to the reference Goat genome. The quantification results produced by the quantifier.pl module were then filtered with a custom perl script parse_miRDeep2_outputs.pl. RNA sequencing data were deposited in the Gene Expression Omnibus (GEO): GSE61025. Statistical anal-ysis to determine differential expression was performed using the Bioconductor package DESeq2.
Results: Food deprivation modulated the expression of 30 miRNA (padj < 0.1); 16 were down-regulated and 14 were up-regu-lated. Diana-microT predictive tools suggested a potential role for several nutriregulated miRNA in lipid metabolism. Among the puta-tive targets, 19 were previously identified as differently expressed genes (DEG). The functions of these 19 DEG revealed, notably, their involvement in tissue remodelling. Some nutriregulated miR were located within QTL region associated with milk trait.
Conclusions: This study offers the evidence of nutriregulated miRNA in the ruminant MG. Characterization of these 30 miRNA could contribute to a clearer understanding of gene regulation in the MG in response to nutrition.
11
Influence of Polymorphisms in Taste
Receptors Genes CD36 and TAS1R2 on the
Metabolic Profile of Obese Children and
Adolescents
M.B. Pioltine1, M.E. Melo1–3, A.E. Fernandes1,
C.T. Fujiwara1, C.M. Brito1,2, C. Gutierrez1, M.B. Prudencio1, A.S. Santos3, C. Cercato2, A. Halpern2, M.C. Mancini1–3 1League of Childhood Obesity, Hospital das Clinicas, Faculty of Medicine, University of São Paulo, São Paulo, Brazil; 2Group of Obesity and Metabolic Syndrome at the Department of Endocrinology and Metabolism, Hospital das Clinicas, Faculty of Medicine, University of São Paulo, São Paulo, Brazil; 3Carbohydrates and Radioimmunoassay Laboratory, Faculty of Medicine, University of São Paulo, São Paulo, Brazil
E-mail: [email protected]
Background: Childhood obesity has a direct impact on quality of life, as well as the future risk of chronic diseases. A dietary pattern rich in fats and sugars and poor in dietary fibers, vitamins and miner-als has contributed to the rise in obesity. Taste is an important
predic-tor of food choices, and polymorphisms in genes encoding taste receptors may explain the individual variability of taste and food preference.
Objective: Our aim was to evaluate the influence of polymor-phisms in taste receptor genes on diet and metabolic profile in obese youth.
Methods: We conducted a cross-sectional study with obese chil-dren and adolescents and a control group of eutrophic chilchil-dren. We genotyped rs1761667 and rs1527483 in CD36, and rs9701796 and rs35874116 in TAS1R2 by real time PCR using Step One Plus® (Applied Biosystems). Food intake was measured by two 24 h food recalls. We evaluated anthropometric and metabolic parameters, including fasting glucose, total cholesterol, HDL-C, LDL-C, triglyc-erides, liver function tests (ALT, AST), insulin, uric acid, leptin and HOMA-IR. Data were analyzed by ANOVA or Kruskal-Wallis test (p < 0.05).
Results: We evaluated 489 obese children and adolescents (Z-BMI 3.29 ± 0.58), and a control group of 130 normal weight chil-dren (Z-BMI –0.17 ± 0.72) 7 to 18 years old. Obese carriers of the minor allele in rs9701796 had lower levels of glucose (p = 0.01), ALT and AST (p < 0.01 for both), suggesting a protective effect of this variant. The minor allele for rs35874116 was associated with a higher frequency of hypertriglyceridemia (p = 0.01). There was no relation between CD36 and TAS1R2 genotypes and food intake. The SNPs rs1761667 and rs1527483 had no effect on the clinical and metabolic profile.
Discussion: At the meantime we found no relationship between genotypes and diet, which could be contributed by the lower frequency of genotypes in some participants. The haplotype analysis will be held whereas we can find a positive association between diet and haplotypes of taste receptor genes, regardless of genotype.
Conclusion: Polymorphisms of TAS1R2 were related to the met-abolic profile of obese children and adolescents. There was no rela-tionship between TAS1R2 and CD36 variants and dietary intake.
12
Resolution Is Delayed in Aged Mice:
Rescue with Nanomedicines
Hildur H. Arnardottir, Jesmond Dalli, Romain A. Colas, Charles N. Serhan
Center for Experimental Therapeutics and Reperfusion Injury, Department of Anesthesiology, Perioperative and Pain Medicine, Harvard Institutes of Medicine, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA 02115, USA
E-mail: [email protected]
Objective: Aging is associated with an overt inflammatory phe-notype and physiological decline. Specialized proresolving lipid mediators (SPMs), namely lipoxins, resolvins, protectins and mares-ins, are autacoids biosynthesized from essential fatty acids that actively promote resolution of inflammation. Here, we investigated endogenous resolution programs in aging and the roles of SPM (Arnardottir et al. 2014. J Immunol 193:4235–4244).
Methods: To investigate resolution pathways in aging we used self-limited acute inflammation in vivo and a systems approach, inte-grating liquid chromatography–tandem mass spectrometry (LC-MS-MS)-based lipid mediator (LM) metabololipidomics, principal component analysis (PCA), and cellular and molecular analyses (e.g. Flow Cytometry and efferocytosis assay). Resolution was quantitated using defined resolution parameters of acute inflammation: Ψmax (maximal neutrophil (PMN) counts), Tmax (the time interval when PMN reach maximum), T50 (the time interval corresponding to 50% PMN reduction, or Ψ50), and Ri (resolution interval, the time interval between Tmax and T50). We also constructed novel nanoproresolving medicines (NPRM) using reprogrammed human monocytes to obtain nanoscaffolds and incorporated SPM for delivery of aspirin-triggered (AT)-Resolvin (Rv) D1 and AT-RvD3 (termed Resolvin-NPRM). These novel Resolvin-NPRM were administered to aged mice (i.p.; 2 × 105 Resolvin-NPRMs containing ~20 ng each of AT-RvD1 and AT-RvD3) either prior to (T0) or 2 h after (T2) inflammatory chal-lenge with zymosan particles (0.1 mg/mouse).
Results: We found that aged mice had an unappreciated aberrant endogenous resolution program with heightened inflammation, delayed resolution and reduced SPM levels, including (LX) B4 (76.6 ± 43.8 vs. 201.5 ± 9.7 pg/mouse), RvD1 (132.6 ± 48.6 vs. 226.1 ± 107.8 pg/mouse), Protectin (PD)1 (19.4 ± 12.8 vs. 33.2 ± 11.8 pg/mouse) and Maresin (MaR) 1 (7.1 ± 3.8 vs. 16.1 ± 8.7 pg/ mouse) compared to matched young mice. Also, aged mice had dys-regulated temporal LM biosynthesis following a self-limited inflam-matory challenge when compared to young mice. The SPM precursor DHA accelerated resolution and promoted human monocyte repro-gramming, actions that correlated with elevated levels of RvD1 and RvD3. Furthermore, RvD3 (50 ng/mouse, i.p.) shortened the Ri by 92% (2 h vs. 23 h in peritonitis plus vehicle mice) and enhanced aged mouse macrophage uptake of apoptotic PMN ex vivo by 47.8 ± 9.5% from vehicle. In aged mice, administration of Resolvin-NPRM lim-ited neutrophil numbers by ~20% and promoted macrophage uptake of apoptotic PMN by 61.5 ± 31.6% and 124.8 ± 51.7% when admin-istered either prior to or 2 h after zymosan challenge, respectively.
Conclusion: These findings provide evidence for age-dependent dysregulation in resolution pathways for acute inflammation and potential new means to activate resolution in an aged innate immune system. Taken together, they provide a new paradigm for improving age-related diseases characterized by excessive innate inflammatory responses.
13
Diet and Methylation of Candidate Genes
Associated with Head and Neck Cancer
Prognosis
E. Anna Arthur1, Shama Virani2, A. Justin Colacino2, L. Emily Bellile3, A. Lisa Peterson4, T. Gregory Wolf4, S. Laura Rozek2,4
1Department of Nutrition Sciences, University of Alabama at Birmingham, Birmingham, AL, USA; 2Department of Environmental Health Sciences, University of Michigan School of Public Health, Ann Arbor, MI, USA; 3Department of Biostatistics, University of Michigan School of Public Health, Ann Arbor, MI, USA; 4Department of
Otolaryngology, University of Michigan Medical School, Ann Arbor, MI, USA
E-mail: [email protected]
Objective: Dietary intake is associated with head and neck can-cer (HNC) prognosis. Epigenetic changes are potentially reversible and are modifiable by environmental factors, including diet. We pre-viously identified that the epigenetic profiles of HNCs are associated with pretreatment dietary intake. Here, we extend these findings, test-ing the hypothesis that diet is associated with candidate gene meth-ylation markers previously shown to be biomarkers of HNC prognosis.
Methods: This study utilized pretreatment data collected from 186 newly diagnosed HNC patients enrolled in the University of Michigan Head and Neck Specialized Program of Research Excellence (UM HN-SPORE). Dietary patterns were derived from food frequency questionnaires (FFQs) using principal component analysis. Promoter methylation of six candidate genes (CCNA1, NDN, CD1A, DCC, p16, GADD45A) was analyzed by pyrosequenc-ing. Multivariable generalized linear models tested associations between each methylation marker and diet variable of interest, cate-gorized into quartiles of intake (whole foods pattern, Western pattern, cruciferous vegetables, green leafy vegetables, refined grains).
Results: After controlling for age, tumor site, HPV-status, smok-ing status, pack years, and comorbidity score, subjects reportsmok-ing in the high quartile, Q4, of green leafy vegetable intake were found to have significantly higher methylation of CD1A compared to those reporting in the lowest quartile, Q1, (p = 0.01) and a test for linear trend indicated significantly higher methylation of CD1A with each increasing quartile (p = 0.03). Subjects within Q4 of the whole foods dietary pattern score had significantly lower methylation of DCC than those in Q1, but the test for linear trend was not statistically sig-nificant.
Conclusion: These results suggest that higher green leafy vege-table consumption is associated with higher methylation levels of CD1A, a marker of immune function previously associated with HNC prognosis in this cohort. Future work should test if diet modifies the relationship between these methylation markers and HNC outcomes.
14
Fatty Acid Binding Protein 2 (Fabp2)
Protects the Small Intestine of Male Mice
from Dietary Saturated Fatty Acid-Induced
Endoplasmic Reticulum (ER) Stress
Zeina Jamaluddine, B. Luis Agellon
School of Dietetics and Human Nutrition, McGill University, Montreal, Quebec, Canada
E-mail: [email protected]
Background: Fabp2, an abundant intracellular lipid binding pro-tein found in the small intestine, shows a preference for binding satu-rated fatty acids such as palmitic acid. The current Western-style diet is rich in palmitic acid, a fatty acid known to induce ER stress. Chronic ER stress may underlie the development of acquired meta-bolic disorders. In this study, we determined if the loss of Fabp2 sen-sitizes the small intestine to ER stress in response to high saturated fat (HSF) diet feeding.
Methods: Wild-type (Fabp2+/+) and Fabp2-deficient (Fabp2–/–) mice, matched by age and sex, were placed on either a low fat or a HSF diet. After 4 weeks, the mice were assessed for plasma lipids and metabolic rate. The abundance of mRNAs for activating transcription factor 4 (ATF4) and spliced x-box protein 1 (Xbp-1s) were measured by quantitative real-time PCR along the longitudinal axis of the small intestine to determine ER stress status.
Results: Plasma lipid concentrations and whole body metabo-lism data indicated that male Fabp2–/– mice on HSF diet were at higher risk of hyperlipidemia, and displayed decreased energy expen-diture compared to Fabp2+/+ male mice. The loss of Fabp2 increased mRNA abundance of ER stress markers ATF4 and Xbp-1s in male mice in response to a HSF diet feeding (P < 0.05). However, in female mice, the loss of Fabp2 did not alter ER stress status in the small intestine.
Discussion: In previous studies, our lab demonstrated that male mice lacking Fabp2 displayed greater sensitivity to high fat feeding compared to female mice lacking Fabp2. In the present study, it is apparent that removal of Fabp2 renders the small intestine of male mice more sensitive to ER stress in response to HSF than the intestine of female mice. The reason for this sex dimorphic sensitivity is under investigation.
Conclusion: The results of this study indicate that the small intestine of male mice requires Fabp2 for the protection against dietary saturated fatty acid-induced ER stress whereas the small intestine of female mice remains resistant to ER stress in the absence of Fabp2.
Funding: This research was funded by a grant from the Canada Research Chairs Program.
15
Dietary Fat Intake Modifies the Association
between rs1799983 NOS3 Polymorphism
and Blood Pressure Levels
Leticia Goni1,2, Marta Cuervo1–3, I. Fermín Milagro1–3, J. Alfredo Martínez1–3
1Department of Nutrition, Food Science and Physiology, University of Navarra, Pamplona, Spain; 2Centre for Nutrition Research, University of Navarra, Pamplona, Spain; 3CIBER Fisiología Obesidad y Nutrición (CIBERobn), Instituto de Salud Carlos III, Madrid, Spain
E-mail: [email protected]
Objective: The aims of the current study were to examine the association between rs1799983 NOS3 genetic variant and blood pres-sure levels, and to evaluate potential gene-diet interactions.
Methods: The study encompassed a total of 705 Caucasian Spanish men and women, who voluntarily attended a nutrigenetic service. Anthropometric and body composition measurements, blood pressure levels, habitual dietary habits and physical activity were examined. Similar to previous studies, fixed increments of 15 mm Hg systolic blood pressure (SBP) and 10 mm Hg diastolic blood pressure (DBP) were added to those individuals with diagnosed hypertension. Oral epithelial cells were collected for the genotyping of rs1799983 NOS3 polymorphism, using Luminex® 100/200TM System.
Results: Of the total population, 283 subjects (40.1%) had the GG genotype (wild type), 324 (46.0%) had the GT genotype and 98 (13.9%) were TT. No statistically differences based on genotype were found for anthropometrics, physical activity and dietary variables. After adjusting for confounder variables (gender, age, body mass index (BMI), smoking and physical activity), carriers of the T allele showed 2.6 mm Hg higher DBP than GG individuals (p 0.009, CI 95% 0.63–4.47). However, no relationship with SBP was found. Saturated fatty acids (SFA) modified the association between the rs1799983 and DBP (p for interaction 0.001). The higher the intake of SFA, the higher the DBP among T allele carriers, while the effect was much lower among GG individuals.
Conclusion: The rs1799983 NOS3 genetic variant was associ-ated with a higher DBP in Southern Europeans. Interestingly, SFA intake modified this association. The study of genetic factors and their interactions with the environment may contribute to individual-ized effective treatments of hypertension.
16
DNA Methylation and Hydroxymethylation
in Severe Obese Women after Bariatric
Surgery
Carolina Ferreira Nicoletti1, Carla Barbosa Nonino1, Bruno Affonso Parenti de Oliveira1,
Marcela Augusta de Souza Pinhel1,
Maria Luisa Mansego2,3, I. Fermin Milagro2–4, Maria Angeles Zulet2–4, J. Alfredo Martínez2–4
1Department of Internal Medicine, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil; 2Department of Nutrition, Food Science and Physiology, University of Navarra, Pamplona, Spain; 3Centre for Nutrition Research, University of Navarra, Pamplona, Spain; 4CIBER Fisiología Obesidad y Nutrición (CIBERobn), Instituto de Salud Carlos III, Madrid, Spain E-mail: [email protected]
Objective: Investigate whether the weight loss induced by bar-iatric surgery may affect DNA global methylation and hydroxymeth-ylation as well the methhydroxymeth-ylation on promoter regions of inflammatory genes such as SERPINE-1 and IL-6.
Methods: This trial included 14 Brazilian obese women (mean age 35.5 ± 10 years) that underwent a hypocaloric diet followed by bariatric surgery (perfomed by Roux-en-Y gastric bypass) in a public hospital. Anthropometric measurements (weight, height, waist cir-cumference and body mass index – BMI) and 12-hours fasting blood samples were collected before and six months after the intervention. 5-hydroxymethylcytosine, LINE-1, SERPINE-1 and IL-6 methylation were assessed in in buffy coat cells with High Resolution Melt using a 7900HT Fast Real-Time PCR System.
Results: An average weight loss of 20.1% of baseline body weight was produced by the intervention after six months, and a reduction in BMI (from 44.6 ± 6.2 to 34.7 ± 5.5 kg/m2, p < 0.001), waist circunference (from 124.5 ± 14.3 to 108.8 ± 13.7 cm, p < 0.001), glucose (from 90 ± 10.9 to 84.4 ± 7 mg/dl, p = 0.004), total cholesterol (from 178.2 ± 37.9 to 160.8 ± 25.1 mg/dl, p = 0.007) and triglycerides (from 128 ± 40 to 89.1 ± 22.3 mg/dl, p = 0.037) levels were found. Hydroxymethylation (from 0.08 ± 0.06 to 0.05 ± 0.03%, p = 0.074), LINE-1 (from 67.7 ± 7.7 to 69.2 ± 6.2%, p = 0.599) and SERPINE-1 (from 17.5 ± 4 to 14.7 ± 7.6%, p = 0.254) methylation levels did not change after weight loss. However, IL-6 methylation decreased after bariatric surgery (from 52.6 ± 6.4 to 49.7 ± 4%, p = 0.043). Also, according to the median of weight loss, baseline SERPINE-1 methylation levels were lower in high responder indi-viduals (15.3 ± 3.4%) as compared to the low-responders (20 ± 3.3%).
Conclusions: Weight loss induced by bariatric surgery did not change global DNA methylation and hydroxymethylation, but decreased the IL-6 methylation. Baseline SERPINE-1 methylation may be a predictor weight loss after bariatric surgery.
17
Integrative Studies for the Identification of
Transcriptomic and Epigenetic Biomarkers
Within a Weight-Loss Program
Mirian Samblas1,2, Maria Luisa Mansego1,2, Maria Angeles Zulet1–3, I. Fermín Milagro1–3, J. Alfredo Martínez 1–3
1Department of Nutrition, Food Science and Physiology, University of Navarra, Pamplona, Spain; 2Centre for Nutrition Research, University of Navarra, Pamplona, Spain; 3CIBER Fisiología Obesidad y Nutrición (CIBERobn), Instituto de Salud Carlos III, Madrid, Spain
E-mail: [email protected]
Objective: Identification of candidate biomarkers that distin-guish individual responses for a weight loss dietary treatment by using the integrative analysis of mRNA expression and DNA meth-ylation arrays.
Methods: The study consisted of a randomized sample of 33 obese people (mean BMI = 35.8 ± 4.7 kg/m2) who participated in the Metabolic Syndrome Reduction in Navarra (RESMENA) retrospec-tive trial. They were classified as low (n = 23) or high responders (n = 10) depending on their weight loss (>8% of basal weight). Peripheral blood mononuclear cells (PBMCs) were isolated from blood samples obtained at the beginning and at endpoint. Extracted DNA was sodium-bisulfite converted and fragmented by controlled enzyme digestion previous hybridization to probes of Infinium Human Methylation450 BeadChip kit (Illumina Inc.). RNA was isolated from white blood cells by TRIzol Reagent for microarray analysis, which was performed with the Illumina Human HT-12 v4 gene expression BeadChip. For the methylation analysis, a significance threshold of p < 0.05 and 5% of methylation variation were used, whereas for the expression study a FDR < 0.05 and a fold change >1.5 were accepted.
Results: A total of 2,102 CpG sites in 1,785 genes showed dif-ferential methylation between low and high responders. In the expres-sion analysis, 156 transcripts were differentially expressed between both groups, 20 of which were downregulated and 136 upregulated in low responders. The integrative analysis of both methylation and expression data identified four genes (CD44, FBXW5, MTSS1 and ITPR1) that appeared differentially methylated and expressed in low responders in comparison with high responders.
Conclusion: In summary, DNA methylation has been suggested as a powerful tool for diagnosis and prognosis. The combination of a high-throughput DNA methylation and expression microarray dataset can be a useful strategy to identify novel genes that might be consid-ered as predictors of the dietary response. Future studies are needed for the validation of these genes as outcome indicators within a weight loss program.
18
High Frequency of the LCT C-13910/G-22018
Lactase Non-Persistence (LNP) Haplotype in
Native But Not in Caucasian Descendant
Population of West Mexico
C. Ojeda-Granados1, A. Panduro1, J.R.R. Pinho2,3, O. Ramos-López1, K. Gleyzer2, F.M. Malta2, S. Roman1 1Department of Molecular Biology in Medicine, Civil Hospital of Guadalajara, ‘Fray Antonio Alcalde’ and Health Sciences Center, University of Guadalajara, Guadalajara, Jalisco, Mexico; 2Hepatology Branch, Department of Gastroenterology, School of Medicine, University of São Paulo, São Paulo, Brazil; 3Department of Clinical Pathology, Hospital Israelita Albert Einstein, São Paulo, Brazil
E-mail: [email protected]
Background and Aim: Polymorphisms C/T-13910 and G/A-22018 of the lactase gene (LCT) are associated with variability in lac-tase persistence (LP) until adulthood. C/C and G/G homozygous respectively are LNP genotypes while the presence of T or A allele are associated with LP, especially in Europeans. Nutritional recom-mendations of Dietary Guidelines are often intended to the general population and do not consider the individual genetic and anthropo-logical background. Mexican population is known to be genetically heterogeneous due to its historical background, therefore we aimed to determine the differences in the frequency of these polymorphisms.
Methods: In a cross-sectional study, 1204 unrelated individuals from populations of West Mexico were analyzed: Native (NAT) groups (86 Nahuas and 95 Huicholes), Caucasian groups (CAU) (32 from Villa Purificacion and 99 from Cuquio, Jalisco), 707 Mestizos from Guadalajara, Jalisco (GDL) and 185 from Tepic, Nayarit (NAY). LCT genotyping was performed by Real-Time PCR System (TaqMan). Hardy-Weinberg equilibrium (HWE), linkage disequilib-rium and haplotype were calculated using Arlequin software (version 3.1). Genetic distances and admixture components were determined by Structure software (version 2.3.4).
Results: Genetic interpopulational differences were observed. The LNP alleles were prevalent in NAT population (C 99.4%/G 99.4%) whereas LP alleles were higher in CAU group (T 28.2%/A 28.2%, p < 0.001). Frequency of C/G LNP haplotype was highest in NAT group (99.4%) and decreased as the European ancestry raised (84.1, 79.3 and 71.8% for NAY, GDL and CAU respectively, p < 0.001). Overall, 68.8% population of West Mexico was LNP. Genotype frequencies were in HWE except NAY group.
Conclusion: Mexicans are genetically heterogeneous. West Mexican population was mostly LNP, however it depended on the ancestral component of each subgroup, as the C/G LNP haplotype decreased as the European ancestry raised. Data presented here will be useful to support regionalized nutritional recommendations by Dietary Guidelines.
19
Fried Foods Might Interact with Genes to
Influence Body Measurements in Women
with History of Gestational Diabetes
Aline Carvalho1,2, Ping Shao3, Huikun Liu3, Junhong Leng3, Weiqin Li3, Leishen Wang3, Shuang Zhang3, Gang Hu3, Lu Qi2
1School of Public Health University of Sao Paulo, Sao Paulo, Brazil; 2Harvard School of Public Health, Boston, EUA; 3Tianjin Women’s and Children’s Health Center, Tianjin, China
E-mail: [email protected]
Background: Obesity has a complex etiology involving behav-ioral, environmental, and genetic factors. Fried food intake was recently found to interact with genetic factors in relation to obesity in Caucasians; however, no study has tested such gene-diet interaction in Chinese population.
Methods: We analyzed the relationship between fried food intake and genetic risk score of BMI among 899 Chinese women with history of gestational diabetes mellitus (GDM). We used a food fre-quency questionnaire to assess fried food intake; and created a genetic risk score on the basis of 10 single nucleotide polymorphisms (SNPs) associated with BMI in East Asians.
Results: Women with a higher genetic risk score had higher weight and waist (P < 0.05). We found that the genetic risk score sig-nificantly interacted with fried food intake in relation to adiposity measures. Participants with a lower genetic risk score had higher dif-ferences in BMI, weight and waist among women who consumed higher fried food than women who consumed lower fried food (dif-ferences for BMI: 2.81 kg/m2; for weight: 6.19 kg; for waist: 7.56 cm), whereas the corresponding differences in those with a higher genetic risk score were 2.24 kg/m2 for BMI, 5.57 kg for weight and 5.24 cm for waist (P for interaction < 0.05 for BMI, weight and waist).
Conclusion: Our data indicate that fried food intake might inter-act with genetic risk finter-actors in relation to body adiposity in Chinese women with history of GDM.
20
Effect of Heterocyclic Amines from Meat
Intake on DNA Damage According to
GSTM1 Polymorphism
Aline Carvalho1,3, Josiane Steluti1, Antonio Carioca1, Gilka Gattar2, Regina Fisberg1, Lu Qi3, Dirce Marchioni1 1School of Public Health, University of Sao Paulo, Sao Paulo, Brazil; 2School of Medicine, University of Sao Paulo, Sao Paulo, Brazil; 3Harvard School of Public Health, Boston, EUA
E-mail: [email protected]
Objective: Heterocyclic amines (HCA) from meat intake are carcinogenic compounds metabolized in the human body. This
pro-cess is mediated by detoxifying enzymes, such as glutathione-S-transferase θ, which is encoded by the GSTM1 gene. The aim of the study was to investigate the relationship between HCA intake and DNA damage, according to GSTM1 genotype.
Methods: A cross-sectional survey (ISA-Capital) was performed among adults and the elderly in Brazil with 73 participants with high HCA intake and 73 gender-and-age-matched participants with non-HCA intake (n = 146). The non-HCA intake was assessed using a 24-hour dietary recall and a structured questionnaire with cooking methods and meat of doneness level information. We used PCR-based assays to detect GSTM1 deletion. DNA damage was measured by 8-oxo-2’-deoxyguanosine (8OHdG) in plasma using an ELISA kit. General linear regression models were used to assess the association between 8OHdG and HCA intake.
Results: The frequency of deletion variant of GSTM1 was 44%. The mean plasma level of 8OHdG concentration was 18.3 ng/ml (95% CI: 16.9; 19.6 ng/ml). The 8OHdG concentration was signifi-cantly associated with HCA intake (β = 1.63 p = 0.040) after adjust-ment for sex, age, race, BMI, C reactive protein, physical activity, smoking, energy intake and fruit and vegetable intake. We found that the association between 8OHdG and HCA was significant among individuals with GSTMT1 wild-type genotype (p = 0.037), but not among individuals with GSTM1 null genotype (p = 0.078).
Conclusion: These findings suggest GSTM1 genotype might modify the relation between HCA and DNA damage.
21
Frequency of Thirteen Single Nucleotide
Polymorphisms Associated with
Susceptibility to Obesity in a Brazilian
Population Sample
T.M.M. Fujii1,2, M.A. Horst3, M. Vilhena1, C. Santos1, A. Reis1, C. Vilhena1, R.S. Diaz1,4
1Centro de Genomas Laboratory, Sao Paulo, Brazil; 2Department of Nutrition, School of Public Health, University of Sao Paulo, Sao Paulo, Brazil; 3Nutrition Faculty, Federal University of Goias, Goiania, Brazil; 4Department of Medicine, Federal University of Sao Paulo, Sao Paulo, Brazil
E-mail: [email protected]; [email protected]
Objective: The purpose of present study was to investigate the frequency of thirteen single nucleotide polymorphism (SNP) associ-ated with susceptibility to obesity in a Brazilian population sample, evaluated by a predictive commercial test, named Nutrigenetica®.
Methods: In this work, 293 people aged 18–83 years partici-pated (187 women and 106 men). We selected patients who did the predictive commercial test since December, 2011 from February, 2015. Briefly, 1 ml of saliva samples was collected in an Oragene DNA (OG-510) saliva collection device and the allelic discrimination experiment was performed in the Taqman OpenArray® Genotyping system in Centro de Genomas® Laboratory, located in Sao Paulo, Brazil. We genotyped thirteen SNP (FTO rs8050136 and rs9939609; MC4R rs10871777, rs12970134 and rs17782313; ADBR2 rs1042714;
LEP rs7799039; TEMEM rs6548238; PLIN1 rs894160; SH2B1 rs7498665; GNPDA2 rs10938397; TCF7L2 rs7903146; PPARG rs1801282). We calculated de minor allele frequency (MAF) for each SNP and then compared with scientific literature. The reference used was the data from HapMap Project, based on CEU population (Utah residents with Northern and Western European Ancestry from the CEPH collection).
Results: All the genetics variants were in Hardy-Weinberg equi-librium. In relation to HapMap data, we observed similar MAF fre-quencies. For FTO (rs8050136 and rs9939609), was A = 0.43/0.46; for MC4R (rs10871777, rs12970134 and rs17782313), was G = 0.23/0.27, A = 0.23/ 0.28, A-0.23/0.26, for ADBR2 was G = 0.32/ 0.47; for LEP was A = 0.41/0.47; for TEMEM was T = 0.20/0.15; for PLIN1 was T = 0.38/0.32; for SH2B1 was T = 0.32/0.38; for GNPDA2 was G = 0.56/0.45; for TCF7L2 was T = 0.38/0.28 and for PPARG was G = 0.07/0.10 for our samples and HapMap data, respec-tively.
Discussion: In Brazil, scientific evidences have been shown the prevalence of overweight in men and women adults is on average 50%. Some lifestyle factors can influence the weight gain and we experimented the increase of saturated fat, sugar and sodium con-sumption, and the decrease of fruits and vegetables ingestion. We are also more sedentary now than years ago. Candidate genes related to obesity are associated to risk for increase body mass index (BMI), waist circumference and waist-to hip ratio, but the risk is different when we compare distinct ethnics groups. Based on Pena and col-leagues (2011) evidences that Brazilian people are more similar with European ancestry, at least, 60% in all Brazilian geographic regions, we compared our results with HapMap data. Since obesogenic envi-ronment can act as a trigger for those who have genetic variant and know the genomic variation can contribute to decrease the risk for development to obesity.
Conclusion: Genetic variants described in this study may con-tribute to future research in the obesity’s field, with the identification of candidate genes associated with this profile in Brazilian popula-tion. May also encourage further intervention studies with the appli-cation of personalized nutrition.
22
Toll Like Receptor 4 Polymorphism Interacts
with Plasma Polyunsaturated Fatty Acids to
Modulate the Risk for Systemic
Inflammation
T.M.M. Fujii1,2, M.M. Norde1, E. Oki1, R.M. Fisberg1, D.M.L. Marchioni1, J.M.P. Souza3, M.M. Rogero1 1Department of Nutrition, School of Public Health, University of São Paulo, São Paulo, SP, Brazil; 2Centro de Genomas Laboratory, São Paulo, SP, Brazil; 3Department of Public Health, School of Public Health of University of São Paulo, São Paulo, SP, Brazil
E-mail: [email protected]
Objective: This study aimed to investigate the interaction between Toll like receptor (TLR) 4 gene single nucleotide polymor-phism (SNP), rs11536889, and plasma polyunsaturated fatty acid
(PUFA) profile on the modulation of risk for systemic inflammation in a cross-sectional population based study.
Methods: Across the participants of Health Survey-São Paulo, we selected all adults (19–59 y), who matched the inclusion criteria to participate in the study (n = 262). Information on smoking habits was obtained using structured questionnaires. Anthropometric and blood-pressure measurements were accessed using standardized procedures. From the blood samples, we analyzed plasma inflammatory biomark-ers levels and FA profile and genotyped TLR4 SNP rs11536889. Hardy-Weinberg equilibrium was accessed using Chi-squared test with continuity correction. A multivariate cluster analysis (MCA), using K-means, was performed to group according to eleven plasma inflammatory biomarkers levels. Patients were separated into two Clusters, representing non-inflammatory (NINF; n = 169) and Inflammatory (INF; n = 93) clusters. Comparisons between clusters were conducted using the Student’s t test and adjusted OR obtained with multiple logistic regression.
Results: SNP rs11536889 was in Hardy-Weinberg equilibrium. Individuals in INF cluster had higher age, waist circumference, sys-tolic blood pressure, diassys-tolic blood pressure, triglyceride levels and smoking habits than subjects in NINF. Individuals with plasma AA and n-3 HUFA levels above the median had lower risk of being in the INF cluster (OR (95% CI) = 0.55 (0.32–0.95) and 0.50 (0.28–0.88), respectively). When stratified by genotypes, a lower risk of being in the INF cluster was found only for GG carriers in the upper percentile of plasma AA, DHA and total n-3 (OR (95% CI) = 0.48 (0.26–0.87), 0.52 (0.28–0.96) and 0.54 (0.29–0.99), respectively). A significant gene-diet interaction was found between rs11536889 and AA/EPA ratio (p = 0.034) and C allele carriers had higher risk of being in the INF cluster when their plasma AA/EPA ratio was above the median (OR (95% CI) = 5.96 (1.02–34.68)).
Conclusion: Inflammatory pattern was related to metabolic syn-drome components, such as hypertension, dyslipidemia and elevated waist circumference, what confirms the contribution of inflammation to metabolic disorders. Elevated plasma AA and n-3 HUFA levels were associated with protection against this inflammatory pattern and TLR4 SNP rs11536889 GG genotype was strongly associated with plasma AA, DHA and n-3 protection. This TLR-4 SNP interacts with plasma AA/EPA ratio to modulate the risk for systemic inflammation.
23
High Consumption of Saturated Fat Intake
Modulates the Effect of FTO on HOMA in
Chilean Population
P.D. Keenan1, C.A. Celis-Morales1–3, S. Abraham1, N.D. Willis1, J.M.R. Gill3, M.E.S. Bailey3, J.C. Mathers1 1Human Nutrition Research Centre, Institute of Cellular Medicine, Newcastle University, Newcastle-upon-Tyne, UK; 2Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK; 3School of Life Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK E-mail: [email protected]
Objective: The genetic and environmental factors contributing to obesity and insulin resistance are widely explored. Genetic varia-tion in the fat mass and obesity-associated gene, FTO, modulates obesity risk and also metabolic markers associated with type 2 diabe-tes mellitus (T2D). However, the effects of interplay between FTO variation and nutrient consumption on disease risk is not well under-stood. In this study we investigated how dietary macronutrients influ-ence the association between FTO genotype and T2D-related metabolic markers.
Methods: We investigated associations between the FTO SNPs, rs8050136 (A/C) and rs17817449 (G/T) and quantitative trait mea-sures in a cross-sectional population sample from Chile (n = 409, 56% females). These quantitative traits included anthropometric (BMI, waist circumference (WC), body fat), metabolic (glucose, insulin, HOMA-IR, lipid profile and liver function), social and life-style variables, including dietary intake and physical activity.
Results: Both FTO SNPs were in perfect linkage disequilibrium, therefore we report the results for the SNP rs17817449 (G/T) only. FTO genotype was significantly associated with body weight (β: 3.1 kg, SE: 0.8 per copy of the risk allele, p = 0.0002), BMI (β: 0.95 kg/m2, SE: 0.2, p = 0.0009), insulin (β: 3.46 pmol/l–1, SE: 0.5, p < 0.0001) and HOMA-IR (β: 0.84, SE:0.13, p = 0.0001) but not rated fat (β: 0.41 g, SE: 0.93, p = 0.657). We found an ‘FTO x satu-rated fat’ interaction effect on HOMA-IR (p = 0.003), such that the effect of FTO genotype on HOMA-IR was significantly greater for those individuals with a higher intake of saturated fat (β: 1.23, SE: 0.20, p < 0.0001) compared to those with lower intake (β: 0.45, SE: 0.20, p = 0.025). All results were adjusted for age, sex, environment, ethnicity, smoking and socio-economic status.
Conclusion: These findings provide evidence that the effects of FTO genotype on T2D risk markers are modulated by saturated fat intake and, specifically, that higher intakes of saturated fat exacerbate the effect of the FTO risk allele on HOMA-IR, a marker of insulin sensitivity.
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Nutrigenetic Factors Associated with Type 2
Diabetes and Obesity in Amerindian
Populations: A Systematic Review
D. Patrick Keenan, Carlos A. Celis-Morales, Ammar Ashor, Jose Lara-Gallegos, D. Naomi Willis, C. John Mathers Human Nutrition Research Centre, Institute of Cellular Medicine, Newcastle University, Campus for Ageing and Vitality, Newcastle-upon-Tyne, NE4 5PL, UK
E-mail: [email protected]
Objective: Western lifestyles have been linked with higher prev-alence of obesity-related non-communicable diseases including type 2 diabetes (T2D). For example, the cultural Westernization of Amerindian populations has been accompanied by increased preva-lence of cardiovascular disease and T2D. Since Amerindian ethnic groups appear to particularly susceptible to non-communicable dis-eases, it is important to investigate the nutritional, environmental and genetic factors which may confer enhanced risk in these populations. To address this gap, we undertook a systematic review of the evi-dence of prevalence of obesity and T2D and of related lifestyle fac-tors among Amerindians.
Methods: Four databases were searched, namely EMBASE, PubMed, Scopus and Cochrane, for relevant articles that could be included in the review. The core search equation was: [Amerindian] AND [type 2 diabetes OR obesity]. The search was specific for human studies. No limits were added to the search. A manual search of the reference lists of relevant articles was also conducted to find any additional papers that were not returned by the initial search. Reviewers scrutinized the returned articles and excluded any studies not resulting in the disease outcomes of interest or pertaining to Amerindian peoples.
Results: The database searches returned a total of 9,473 titles and abstracts. After removal of duplicates, 4,623 abstracts were left for content review. After further review, 145 articles met the inclusion criteria and are the focus of the review.
Conclusion: This systematic review, despite having no search limits, has shown a surprising lack of diversity in the Amerindian populations studied. This demonstrates the need for more research in a broader range of populations to fully understand the relationship between nutrigenetic factors, T2D and obesity in Amerindians and to strengthen the evidence base for clinical application.
