Parte de la presentación de mi tesis doctoral (2010) con el titulo " Molecular mechanisms involved in the protective effect of the Mediterranean diet and olive oil consumption in humans".

1. 1
Designed by Júlia Muñoz Carreras

2. Molecular mechanisms involved in
the protective effect of the
Mediterranean diet and olive oil
consumption in humans
Valentini Konstantinidou, PhD
Thesis director: Dr. Maria Isabel Covas Planells
Cardiovascular Risk and Nutrition Group,
Institut Municipal d´Investigació Mèdica (IMIM-Hospital del Mar)
PhD in Biomedicine, Universitat Pompeu Fabra,
Department of Experimental and Health Sciences,
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Barcelona, Spain

3. Presentation Outline
 Introduction in Nutrigenomics
 Objective-Hypothesis
 Methods and Results
 Conclusions
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4. Introduction – Nutrigenomic era
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5. Introduction – Nutrigenomics
Principles
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Müller and Kersten, Nature Reviews 2003

6. Introduction – Nutritional Genomics
 Nutrigenomic studies have focused on investigating the
molecular mechanisms of action of several foods and nutrients,
particularly lipids, on cardiovascular risk factors and other
complex traits.
 Intervention studies, in which subjects receive a controlled
dietary intake, provide the best approach for conducting cause-
effect relationships between gene expression and diet.
Limitations:
- Small number of participants
- Brief intervention duration
- Lack of replication
Ordovas et al. Mol Nutr Food Res 2007
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7. Introduction – Nutrigenomic studies general
Goals
 To understand the molecular mechanisms of genes-diet interaction.
 To prevent diet-related diseases.
 To develop evidence-based nutrition.
 To contribute to public health.
State of the art
 Mechanisms by which dietary patterns, foods or food components elicit their
beneficial/harmful effects on human health are partially unknown.
 One of the mechanisms could be the modulation of disease-related genes by
dietary patterns, foods or food components.
 At present, the knowledge concerning the role of diet in modulating
atherosclerosis-related genes is limited.
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8. Introduction – Nutrigenomic era
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9. Introduction – Nutrigenomic studies II
In peripheral blood mononuclear cells (PBMNCs)
• Higher increase in TNFα expression after butter-rich breakfast vs olive oil.
• Higher increase in IL6 mRNA response after butter-rich breakfast vs walnut.
(Jimenez-Gomez et al. Atherosclerosis 2009)
• Postprandial activation of NFκ B after butter- and walnut-rich meals vs olive oil.
(Bellido et al. Am J Clin Nutr 2004)
In human adipocytes
• Anti-inflammatory gene expression profile after MUFA-diet vs SFA-diet.
(van Dijk et al. Am J Clin Nutr 2009)
• Higher TNFα expression after conjugated linoleic acid (CLA) vs olive oil.
• Lower expression in GLUT4 (glucose transporter 4), LEPTIN, LPL (lipoprotein
lipase) after CLA vs olive oil.
(Raff et al. J Nutr 2009)
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10. Hypothesis
 The consumption of a TMD, VOO and its
PC, can modify the human in vivo gene
expression.
 The gene expression changes will be
towards a protective mode for
cardiovascular disease development.
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11. Methods - Task 1: VOO intervention study-pilot (n=11)
Design : Linear Study
Wash out period
1-4 days: habitual diet 5-7 days: diet with very
controlling excess of low phenolic content,
antioxidants, sunflower sunflower oil for raw
oil for raw and cooking and cooking purposes
purposes
• Intervention day
Samples collection 0h 1h 6h
50ml of VOO
ingestion
• 3 weeks (Khymenets et al. OMICS 2009)
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12. Results - Task 1: VOO intervention study
Characterization of Human Gene Expression Mononuclear Transcriptome
Changes after Olive Oil Ingestion: an Response after Sustained Virgin
Exploratory Approach. Olive oil Consumption in humans
Konstantinidou, V. et al. (2009) Khymenets, O. et al. (2009)
Folia Biologica (Praha) 55, 85-91 OMICS 13, 7-19
7 insulin sensitivity-related genes were modulated by VOO ingestion
Task 1.2
Postprandial time course of changes in the expression of those genes
after VOO ingestion (qRT-PCR)
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13. Methods
Characterization of Human Gene Mononuclear Transcriptome Response
Expression Changes after Olive Oil after Sustained Virgin Olive oil
Ingestion: an Exploratory Approach. Consumption in humans.
Konstantinidou et al. Khymenets et al. OMICS 2009
Folia Biologica (Praha) 2009
Time Course of Changes in the Expression of
Insulin-Sensitivity Related Genes after an Acute
Load of Virgin Olive Oil.
Konstantinidou, V. et al.
OMICS 2009
Literature Selection of 47
review atherosclerosis-related genes
Task 2: TMD intervention
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14. Methods - Task 2: TMD intervention study
Study Flow diagram
Invited to be screened
(n=99)
Ineligible (n=9)
Did not meet inclusion criteria (n=7)
Declined to participate (n=2)
Randomly assigned
(n=90)
Declined to follow
up (n=1)
Control Group (n=29) TMD+WOO (n=30) TMD + VOO (n=30)
(n=20 in gene expression) (n=16 in gene expression) (n=20 in gene expression)
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15. Results - Task 2: TMD intervention study, Three-group analyses
Gene expression changes after TMD+VOO intervention
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p<0.05 for linear trend in all cases * p<0.05 vs. control

16. Results - Task 2: TMD intervention study
Volunteer’s dietary compliance
• Volunteers compliance was good as it is reflected in urinary T and HT levels
† †
p=0.007 for quadratic trend
Hydroxytyrosol (ng/ml)
Tyrosol (ng/ml)
*
Control group TMD+WOO TMD+VOO Control group TMD+WOO TMD+VOO
* p<0.05 vs control; † p<0.05 vs TMD+WOO 16

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18. Conclusions
Strengths
 To work in real life conditions in all interventions
 To use both the whole dietary pattern and the single component approach
 To work against our hypothesis by using a Spanish dietary pattern in the control group
Limitations
- A lack of control group for the VOO intervention
- The inability to assess potential interactions
- As expected, we observed modest changes in gene expression
- Unknown effects over longer intervention periods
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19. Conslucions
IFNγ ADAM17
OGT
IL7R ALOX5AP
ARHGAP15
LIAS
ADRB2
CD36
PPARBP
POLK
Inflammation Oxidative stress Insulin resistance
The gene expression changes were observed in a protective
mode for counteracting these situations.
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20. Discussion
These results provide, for the first time, evidence on:
 An in vivo human nutrigenomic effects of the TMD, in healthy
volunteers.
 An in vivo human nutrigenomic effect of olive oil phenolic
compounds down-regulating atherosclerosis-related genes.
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21. Thank you for your attention
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