Curriculum vitae

2002-2005 Assistant Researcher

Project "Response in calcitonin therapy and association with CTR polymorphisms in osteoporotic postmenopausal women". " Development of a system for the management and analysis of biological samples and "-omics" data related to VEGF", 7000 euros, 1 year duration

Medical

List of Publications

Original publications 1.

Ligthart S, Vaez A, Võsa U, Stathopoulou MG, ... , Alizadeh BZ. Genome-wide association analyses of >200,000 individuals identify 58 genetic loci for chronic inflammation and highlights pathways that link inflammation and complex disorders. Am J Hum Genet.

2018 Nov 1;103 (5):691-706.

Organisation of conferences

1) The 9th Santorini Conference, Santorini, Greece, 2018.

2) 8th Santorini Conference, Santorini, Greece, 2016.

3) Third ESPT Conference: Integration of pharmacogenomics in clinical decision support, Budapest, Hungary, 2015.

4) 7th Santorini Conference Biologie Prospective, Santorini, Greece, 2014.

Résultats attendus

Les principaux résultats attendus sont:

• Déterminants «-omiques» du VEGF à utiliser pour la détermination moléculaire des MCV, de l'AD et de l'ostéoporose.

• Nouveaux modèles de prédiction du risque basés sur les «-omiques» du VEGF -algorithmes pour les MCV, l'AD et l'ostéoporose.

Ces résultats auront un impact important sur la connaissance de la biologie du VEGF et de son implication dans les maladies évaluées, mais démontreront également son utilité en tant que biomarqueur de la médecine personnalisée grâce à la prédiction précoce du risque.

Les connaissances générées par le projet seront mises à la disposition de la communauté scientifique et du public à la suite de plusieurs publications dans des revues internationales à facteur d'impact élevé dans les domaines médical, génétique et épidémiologique et lors de conférences scientifiques.

Résumé des activités de valorisation et de transfert et visibilité scientifique en francais

Nombre total de publications (revues à comité de lecture): 66 heritability estimated to be 50-80%) 11 and, as most chronic diseases, it is characterized by polygenic regulation 12 . Both candidate genes and genome-wide associations studies (GWASs) have been successful in the identification of genetic variants associated with BMD and they have also provided novel biological pathways which led to new therapeutic agents 13 .

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The aim of my master and doctoral research was the assessment of genetic variants associated with lumbar spine and femoral neck BMD, taking into account possible interactions with nutrients known to affect the traits, especially calcium 14 . Selected candidate genes implicated in classical and novel osteoporosis pathways were tested including the vitamin D receptor (VDR), the calcitonin receptor (CTR) and the low-density lipoprotein receptor-related protein 5 (LRP5).

The first study was a cross-sectional study conducted on a sample of 301 healthy young men aiming to assess the genetic effects on peak bone mass. The population aged 18-25 years was selected during gradational examinations for the Greek Armed forces. Distal bone mineral content (dBMC), distal BMD (dBMD) and ultradistal BMD (UdBMD) at the forearm were measured by single X-ray absorptiometry (Osteometer DTX-100, Denmark). Additionally, there were assessments of the corresponding geometrical areas, such as radial area (ArRad), ulnar area (ArUlna) and ultradistal area (ArUd). The polymorphisms rs1801197 of the CTR gene and the rs1544410 of the VDR gene were genotyped. Direct and epistatic interactions between the two polymorphisms were assessed. We found no significant direct association or any epistatic effect between forearm bone mass variables and geometrical areas and the two polymorphisms. This study indicated that these two polymorphisms of known candidate genes for osteoporosis are not determining the peak bone mass in Greek males (original publication 42) 15 . In this work, I was involved in the study design, DNA samples preparation and genotyping, databases preparation and statistical analysis and manuscript revision. The second study was a cross-sectional study on 600 postmenopausal women aiming to assess the genetic effects on bone deterioration during menopause. The subjects were unselected postmenopausal Caucasian women of Greek origin who were asked to voluntarily participate in the study through advertisements in four randomly selected Centers of Open Protection for the Elderly in the Athens region (community centers that aimed for primary health prevention and social support of elderly persons). The BMD of the lumbar spine (L2-L4) and hip was measured using dual-energy X-ray absorptiometry with the Lunar DPX-MD device. The genotyping was performed for the polymorphisms rs1784235, rs491347, rs4988321, and rs4988330 of the LRP5 gene and the rs1544410 (Bsm-I), rs11568820 (Cdx-2), and rs731236

(Taq-I) polymorphisms of the VDR gene. For the polymorphisms of the LRP5 gene, we found that rs4988321 was associated with lumbar spine BMD (P-value=0.002). We also showed a significant interaction of the rs4988321 polymorphism with calcium intake (P-value=0.016),

where the carriers of the A allele had lower lumbar spine BMD compared to GG homozygotes (P-value=0.001) only in the lowest calcium intake group (<680mg/day), while in the highest calcium intake group no differences were found in BMD between genotypes (figure 1). For and between Cdx-2 polymorphism and lumbar spine BMD (P=.025) (figure 2) only in the group of low calcium intake (<680 mg/day). In the higher calcium intake group, the polymorphisms of VDR gene were not associated with the assessed traits. b Percentiles derived using the median value of calcium intake (680 mg/day) as a cutoff point.

(Calcium intake was assessed using two 24-hour recalls and values were averaged).

*** P-value=0.001 for the comparison of mean adjusted spine bone mineral density between rs4988321 genotypes in the <680 mg/day calcium intake percentile (univariate analysis of variance using age, weight, height as covariates).

Figure 2. The effect of Cdx-2 genotypes on spine BMD according to calcium intake.

These findings demonstrated that the polymorphisms of both candidate genes have an important effect on BMD only when calcium intake is low, while higher calcium intake has a protective effect on BMD against genetic determination. These results also highlighted the importance of taking into account the nutritional intake and interactions with genetic polymorphisms when assessing the risk of chronic multifactorial diseases like osteoporosis.

The related results were published in two original publications (original publications 34 and 35) 16,17 . In this work, I was involved in the study design, recruitment of the population, data collection, BMD measurements, DNA samples preparation, dietary analysis, databases preparation, statistical analysis and drafting of the manuscripts. They were presented in 5 conferences (conferences 24, 25, 26, 27, 28)

Project 1 : Study of vascular endothelial growth factors (VEGF) and its involvement in cardiovascular diseases (CVD)

The VEGF-A (most commonly VEGF) is one of the most important factors associated with angiogenesis and is considered a key regulatory molecule for both physiological and pathological angiogenesis 19,20 . Physiological angiogenesis occurs during fetal development.

In the adult, angiogenesis is involved in skeletal growth, pregnancy, wound healing and also in inflammation, tissue graft and ischemia 21 . VEGF, through its two receptors VEGFR-1 and VEGFR-2 (vascular endothelial growth factor receptor 1 and 2) and related signaling pathways, exerts a variety of functions. These include the activation, migration and proliferation of endothelial cells (ECs), the survival of ECs (through enthothelial nitric oxide (NO) production) and the mediation of vascular permeability, thus resulting in angiogenesis and other hemodynamic effects 22,23 (figure 3). Blood vessels are beneficial for normal tissue growth and regeneration but deviations in angiogenesis are connected to various disease.

Insufficient vessel growth results in CVD (stroke, myocardial infarction), ulcerative disorders and neurodegeneration, while increased angiogenesis has been observed in cancer, and inflammatory disease 24 . Especially for cancer, angiogenesis is a fundamental requirement for tumor growth as oxygen and nutrients supply is required for tumor progression 25 . It has been shown that up to 60% of human tumors express the VEGF gene 26 . Due to its important role in angiogenesis, VEGF has been extensively studied and has been consequently associated with a variety of diseases such as those mentioned above. An important application of VEGF biology studies is the production of VEGF inhibitors as therapeutic approach in cancer.

However, non-responsiveness in some type of cancers poses questions concerning VEGF mode of action. Furthermore, significant side effects have been observed with anti-VEGF treatment including severe cardiological effects such as hypertension, cardiac ischemia, arterial thromboembolic events (e.g cerebral and myocardial infarction) and left ventricular dysfunction. Even if the mechanisms for the cardiological side effects of these therapies are not yet well-defined, this observation further supports the connection of VEGF with CVD 21 . VEGF therapies have been tested in randomized clinical trials, particularly in patients with coronary artery disease, with low clinical effects 28 . Higher levels of VEGF have been observed in patients with vascular diseases; nevertheless, it is not clear whether the up-regulation of VEGF is an adaptation to ischemia or if it is rather causal in the onset of these diseases [29][30][31][32][33][34][35][36] . Also, the involvement of VEGF in atherosclerotic process is conflicting. A possible neo-vascularization and proinflammatory effect of VEGF has been identified in animal models and in vitro studies, which leads to progression of atherosclerosis and plaque instability [37][38][39][40][41][42] . In contrary, clinical trials using VEGF in humans 28 and gene therapy in animal studies 43,44 do not support a positive effect of VEGF on atherosclerosis progression. It seems that the relationship between VEGF and CVD is complicated due to the complexity of these multifactorial diseases but also due to pleiotropic effects of VEGF.

VEGF gene gives rise to more than 14 isoforms grouped in two major families due to differential pre-mRNA splicing (figure 4). The VEGFxxx family results from splicing in exons 6, 7 and the proximal splice site in exon 8 (exon 8a), while the VEGFxxxb family isoforms differ in the selection of a distal splice site at exon 8 (exon8b) located 66bp downstream of the proximal splice site. Thus, the two families have the same number of amino-acids, but differ in amino-acid sequence of 6 amino-acids at the C-terminus [START_REF] Nowak | Regulation of vascular endothelial growth factor (vegf) splicing from pro-angiogenic to anti-angiogenic isoforms: A novel therapeutic strategy for angiogenesis[END_REF][START_REF] Woolard | Molecular diversity of vegf-a as a regulator of its biological activity[END_REF] . This variation results in significant differentiation in structure, receptor interaction and function [START_REF] Harper | Vegf-a splicing: The key to anti-angiogenic therapeutics?[END_REF] .

As a consequence, while the VEGFxxx isoforms have pro-angiogenic actions (promotion of angiogenesis), the VEGFxxxb isoforms are anti-angiogenic [START_REF] Nowak | Regulation of vascular endothelial growth factor (vegf) splicing from pro-angiogenic to anti-angiogenic isoforms: A novel therapeutic strategy for angiogenesis[END_REF][START_REF] Harper | Vegf-a splicing: The key to anti-angiogenic therapeutics?[END_REF] . In normal conditions in adults the anti-angiogenic isoforms are expressed in non-angiogenic tissues and represent >50% of total VEGF protein 25 . In contrary, many cancers are characterized by a switch from the anti-angiogenic to pro-angiogenic isoforms which leads to neo-vascularization and consequently to tumor growth 25,[START_REF] Woolard | Molecular diversity of vegf-a as a regulator of its biological activity[END_REF][START_REF] Harper | Vegf-a splicing: The key to anti-angiogenic therapeutics?[END_REF] . Also, it has been suggested that the isoform profile of different tumors may affect the responsiveness to anti-VEGF therapies [START_REF] Woolard | Molecular diversity of vegf-a as a regulator of its biological activity[END_REF][START_REF] Harper | Vegf-a splicing: The key to anti-angiogenic therapeutics?[END_REF] . Another interesting point is that the alternative splicing leading to the different isoforms can be dependent on environmental factors. For example, it is known that VEGF gene is regulated by inflammation molecules (e.g. IGF-1, TNF-α, TGF-β) but there are no data concerning the type of isoforms produced 25 . The understanding of the differential and tissue-specific functions of VEGF related to the different isoforms production in normal and pathological tissues is expected to allow the development of personalized therapies [START_REF] Ng | Vegf function in vascular pathogenesis[END_REF] . Given the complexity of VEGF biology and its importance to the development and therapy of complex diseases including CVD and cancer, this molecule has been investigated by the IGE-PCV; U1122 unit by a detailed integrative approach. In fact, it has designed and implemented a strategy for the study of VEGF and its links with CVD, which uses genetic and transcriptomics methodologies. The additive genetic heritability of VEGF plasma levels has been estimated in a sample of families from the STANISLAS Family Study (SFS) to be 60.5% of the total variance of the trait [START_REF] Berrahmoune | Heritability for plasma vegf concentration in the stanislas family study[END_REF] . Basic biological determinant and reference values have also been established, indicating the age, the platelet count and the alanine aminotransferase activity as important factors in children and adults [START_REF] Berrahmoune | Biological determinants of and reference values for plasma interleukin-8, monocyte chemoattractant protein-1, epidermal growth factor, and vascular endothelial growth factor: Results from the stanislas cohort[END_REF] . Furthermore, a study on pre-analytical variations influence on VEGF blood levels and gene expression has been performed in order to specify the optimal conditions and biological samples for the investigation of VEGF biology in blood [START_REF] Azimi-Nezhad | Influence of pre-analytical variables on vefga gene expression and circulating protein concentrations[END_REF] . The following step included a large GWAS, where a sample of families from the SFS participated for the identification of genetic determinants of VEGF blood levels. Four single nucleotide polymorphisms (SNPs) in 3 chromosomes were significantly associated with VEGF concentrations and explained ~ 50% of the heritability of this phenotype [START_REF] Debette | Identification of cis-and trans-acting genetic variants explaining up to half the variation in circulating vascular endothelial growth factor levels[END_REF] . In the same study, a transcriptomics approach performed by IGE-PCV;

U1122 unit with peripheral blood mononuclear cells (PBMCs) of the SFS individuals identified also six SNPs significantly associated with VEGF mRNA _121 levels, while SNPs annotation and a biological pathway analysis software produced a putative gene network that describes the functional relationships between VEGF gene and the identified SNPs [START_REF] Debette | Identification of cis-and trans-acting genetic variants explaining up to half the variation in circulating vascular endothelial growth factor levels[END_REF] .

Given the important proportion of explained VEGF heritability by the above identified SNPs, in my post-doctoral project we have selected these polymorphisms for the in-depth investigation of possible molecular links between VEGF and other CVD endophenotypes in order to examine physiological pathways that jointly regulate CVD development. I was the principal researcher of these projects.

In the first study, we have assessed the direct effects, gene × gene and gene × environment interactions of the 4 GWAS-identified SNPs on blood lipid levels (apolipoprotein E (apoE), triglycerides, total cholesterol, low-and high-density lipoprotein cholesterol levels (LDL and HDL)]. The study was performed on a discovery population (n=1006) from SFS and a replication population (n=1145) of European origin that makes part of the BRC IGE-PCV (BB-0033-00051). Meta-analysis of the 2 populations was performed using a weighted inverse normal method with a fixed effect using the "META" R package. We have demonstrated that the SNP rs6921438 was associated with HDL (beta = -0.08, Poverall = 1.2 × 10 -7 ) and LDL (beta = 0.13, Poverall = 1.5 × 10 -4 ). We also identified a significant association between the interaction rs4416670×hypertension and apoE levels (Poverall = 1.7 × 10 -5 ). The SNP rs6921438 has been previously associated negatively with levels of VEGF and is the variation that explains 41% of VEGF levels [START_REF] Debette | Identification of cis-and trans-acting genetic variants explaining up to half the variation in circulating vascular endothelial growth factor levels[END_REF] . Therefore, we proposed that the rs6921438 could negatively affect the cardiovascular system through a decrease in HDL-C levels, an increase in LDL-C levels and a decrease in VEGF levels. These results were presented in one original publication [START_REF] Stathopoulou | A common variant highly associated with plasma vegfa levels also contributes to the variation of both ldlc and hdl-c[END_REF] (original publication 28).

In a second study, we have assessed the associations between levels of VEGF and cellular adhesion molecules (CAMs) and inflammation markers, between their expression levels in mRNA (beta = 0.353, P = 0.021), L-selectin mRNA (beta = 0.611, P = 0.001) and TNF-α mRNA (beta = 0.616, P = 0.001). Furthermore, significant epistatic interactions were identified for plasma VEGF, E-selectin, TNF-α, ICAM-1 and IL6 levels, while the SNP rs4416670 was associated with mRNA levels of L-selectin (beta = 0.196, P = 0.039). These results supported the hypothesis of complex and multilevel relationships between VEGF and CAM and inflammation molecules that are present even in healthy states. This work was presented in one original publication [START_REF] Azimi-Nezhad | Associations of vascular endothelial growth factor (vegf) with adhesion and inflammation molecules in a healthy population[END_REF] (original publication 29).

My contribution in these studies involved the studies design, the statistical analysis, the interpretation of the results and the drafting of the original research papers. These two studies were part of the doctoral thesis of Mohsen Azimi-Nezhad, which I co-supervised with Dr Siest (doctoral student 6) and of Marc Rancier (doctoral student 4) and Abdelsalam Saleh (doctoral student 5).

In a third study, the associations of the rs6921438 and rs10738760 VEGF-related SNPs with the risk of type 2 diabetes, nephropathy and retinopathy were assessed. Two case-control populations for type 2 diabetes were used and one case-control population for nephropathy and one for retinopathy were also assessed. No significant associations were identified thus indicating that the links between diabetes and its comorbidities with VEGF are mediated through other mechanisms. This work is presented in one original publication 55 (original publication 26).

Two more doctoral theses co-supervised by me and Dr Siest were initiated during my postdoctoral period on VEGF projects (doctoral students 4 and 5). Their results will be developed in the next section (Research Engineer Inserm).

These results and the developed strategies were at the origins of the "Vascular endothelial growth factor's European Genomic Federation (VEGF)" project that was submitted in 2014 under the HORIZON 2020 PHC-05-2014 «Health promotion and disease prevention:

translating "omics" into stratified approaches» call. The project was coordinated by Dr Siest and I was the project manager. The VEGF Project included 12 participants coming from 6

European Universities and one from the USA. This project aimed to develop and assess VEGF-based personalised prevention programs for CVD integrating "-omics" profiles of individuals, complemented by environmental and lifestyle factors. The project was selected for a second phase evaluation but did not finally receive a funding.

Project 2: Assessment of gene × gene and gene × environment interactions for the regulation of blood pressure

It is well-accepted that hypertension is a major risk factor for CVD with a significant worldwide burden 56 . Although large studies have tried to identify the most significant genetic components of blood pressure, only a very small variance of the relative traits is explained 57 .

The IGE-PCV; U1122 unit has set a strategy for the assessment of blood pressure genetic variants using a combination of candidate genes and GWASs methodologies, also taking into account gene × gene and gene × environment interactions with the application of functional approaches through transcriptomic analyses.

In one study, we have assessed the epistatic interactions between several candidate genes for blood pressure in a discovery population of 3600 individuals not treated with hypertensive therapies and a replication cohort of 4620 subjects. We have identified that the interaction between rs5355 of SELE gene and rs6046 of F7 gene were significantly associated with systolic blood pressure levels in both populations (P= 0.047 and P= 0.03 respectively). An investigation of the underlying mechanisms for these associations was performed through transcriptomic analyses in PBMCs of 90 individuals. The rs6046A allele was associated with increased NAMPT gene mRNA levels (P≤9.2×10 -5 ). NAMPT gene mRNA levels were also positively correlated with ICAM-1, SELL, FPR1, DEFA1-3 and LL-37 genes expression (P≤5×10 -3 ), while DEFA1-3 and LL-37 gene expression levels were associated with systolic blood pressure (P = 3×10 -3 and P = 0.01 respectively. These results suggest an indirect link between NAMPT gene expression and blood pressure through the expression of adhesion and innate immune system molecules, where the identified epistasis plays a role. The findings are presented in one publication 58 (original publication 32).

In a second study, we assessed the epistatic interactions between 100 SNPs 65 candidate genes for blood pressure in a discovery population of 1912 French unrelated adults and an independent replication population of 1755 unrelated European adults. Nine SNPs in 8 genes were significantly associated with blood pressure: LPL and LDLR (lipid metabolism), F7

(coagulation factor), AGT1R (BP regulation), CSF2, IL1B, TGFB1 and VDR (inflammatory pathways) genes. Two significant epistatic interactions were identified for both systolic and diastolic blood pressure in the discovery set: VCAM1 (rs1041163) × APOB (rs1367117) and SCGB1A1 (rs3741240) × LPL (rs1800590) (P≤0.04 and P≤0.03 respectively), while only the effects on diastolic blood pressure were significant in the replication population (P=0.045 and P=0.05 respectively). This study provided novel original results based on epistasis between known candidate genes for blood pressure regulation. These results were presented in one publication 59 (original publication 27) and in the 7th Santorini Conference Biologie Prospective as a poster presentation (conference 11).

My participation for these projects was on the design of the study, statistical analysis and the interpretation of the results as well as the critical correction of the manuscript.

Project 3: A genome-wide association study for the genetic determinants of circulating haptoglobin levels

Haptoglobin is an acute phase inflammatory molecule, whose physiological role is the scavenging of circulation haemoglobin and its degradation in the liver, and it serves as a reverse transporter of cholesterol to the liver. Its levels as well as its polymorphisms have been associated with CVD, other non-communicable diseases and infectious diseases.

In a GWAS study we assessed the genetic variants influencing the levels of haptoglobin in unrelated children. A discovery population of 631 French children from the SFS and 3 replication cohorts (total n=4391) were used. The polymorphism rs2000999 on HP gene was strongly associated with haptoglobin levels in the meta-analysis (P= 8.09×10 -59 ). This SNP was significantly associated also with total cholesterol (β = -0.011, P =0.002) and LDL levels (β = -0.017, P = 0.0008) in 5 independent children cohorts (total n=8789), thus linking haptoglobin levels with CVD risk. These results were published as an original article [START_REF] Froguel | A genome-wide association study identifies rs2000999 as a strong genetic determinant of circulating haptoglobin levels[END_REF] (original publication 31) and in the 6th Santorini Conference Biologie Prospective as a poster presentation (conference 21). My participation in this project was focused on data interpretation and review of the manuscript and it was an excellent occasion for further training in the GWAS methodology and analysis.

Project 4: Telomere length genetic determinants in adults and children

Telomeres are structures made of tandem repeats of hexanucleotide units with numerous telomeric-interacting proteins that are present at the ends of chromosome. They are responsible for the protection of genomic integrity. The loss of several base pairs from telomeres during each mitotic division leads to telomere shortening which, after a critical limit, results to telomere dysfunction that triggers cell cycle arrest, apoptosis and/or cellular senescence [START_REF] Armanios | Telomeres and age-related disease: How telomere biology informs clinical paradigms[END_REF][START_REF] De Jesus | Assessing cell and organ senescence biomarkers[END_REF] . The telomere length has been linked with a variety of human diseases, ranging from rare monogenic disorders to age-related complex diseases such as cancer, CVD and CVD risk factors such as obesity, hypertension and diabetes [START_REF] Armanios | The telomere syndromes[END_REF][START_REF] Meyer | Telomere length and cardiovascular aging: The means to the ends?[END_REF] . An important number of studies have identified genetic variants associated with leukocyte telomere length (LTL) 65- 71 . These studies are focused on adult populations, even though childhood is an extremely crucial period for the determination of LTL and the assessment of age-specific genetic determinants could be of great importance.

I have designed and analysed the data analysis in a study focused on genetic determinants of LTL in children. The effect of 19 previously identified polymorphisms of LTL in adults in GWAS studies was assessed in a sample of 322 unrelated normal weight children from the SFS. None of these variants were significant in our paediatric population, thus a discovery only GWAS approach was used and 6 novel variants, specific for children, reached the levels of suggestive association (P ≤ 5 ×10 -5 ). This study pointed out the importance of studying the genetic background of LTL in children and the preliminary results were presented in one publication [START_REF] Stathopoulou | Genetic determinants of leucocyte telomere length in children: A neglected and challenging field[END_REF] (original publication 22) and in 2 international conferences as poster presentations (conferences 13 and 10). This work was included in the master thesis of Alexandros Petrelis, which I co-supervised (master student 1).

Furthermore, apart from the observed relationships with CVD endophenotypes (e.g. inflammation markers, glucose metabolism, oxidative stress) telomere length has been identified as an independent predictor of heart disease related events [START_REF] Brouilette | Telomere length, risk of coronary heart disease, and statin treatment in the west of scotland primary prevention study: A nested case-control study[END_REF] and thus suggesting its possible use as new biomarker for CVD [START_REF] Meyer | Telomere length and cardiovascular aging: The means to the ends?[END_REF] . It is accepted that telomere length reflects the accumulative burden of inflammation [START_REF] Aviv | Human telomere biology: Pitfalls of moving from the laboratory to epidemiology[END_REF] . Since 2013, I have participated in the "ANR Programme de Recherche Translationnelle en Santé", entitled "TELARTA (Telomeres and Arterial Aging)", where the IGE-PCV; U1122 unit was responsible for the task 5:

"Characterization of inflammation markers in blood samples" for the assessment of plasma inflammation markers' relationships with telomere length of muscular cells, white blood cells and endothelial progenitor cells from CVD patients and healthy subjects. Among the inflammation markers, CRP, TNF-α and IL6 are those most significantly associated with telomere length [START_REF] Bekaert | Telomere length and cardiovascular risk factors in a middle-aged population free of overt cardiovascular disease[END_REF] . However, a wider range of pro-and anti-inflammatory markers have been quantified including VEGF, interleukins 1b, 2, 4, 8, 10 and 12, interferon-γ (IFN-γ), monocyte chemotactic protein-1 (MCP-1), ICAM-1 and vascular adhesion molecule-1 (VCAM-1). My participation included the selection of inflammation molecules and the interpretation of the results, which are ongoing. The assessment of these relationships will increase our knowledge on inflammation and VEGF role in telomere dynamics, as the selected tissues reflect them.

Project 5: Pharmagenomic projects of clopidogrel

Another subject that gathers great attention in the application of genomics in diseases is the field of pharmacogenomics, which studies the effect of genetic variations to drug response [START_REF] Wang | Genomics and drug response[END_REF] .

This field, more than any other, is aiming to personalised medicine. Concerning cardiovascular medications, warfarin and clopidogrel [START_REF] Hayden | Cardiovascular disease gets personal[END_REF] as well as statins [START_REF] Postmus | Pharmacogenetics of statins: Achievements, whole-genome analyses and future perspectives[END_REF] are the most commonly studied medications for possible gene × drugs interactions. The obtained results of pharmacogenomics in CVD have led to clinical considerations. For example, the Food and Drug Administration (FDA) officially suggests that genotypes for CYP2C9 and VKORC1 and for CYP2C19 genes should be taken into account when prescribing warfarin and clopidogrel respectively [START_REF] Wang | Genomics and drug response[END_REF] . Although the progress is slow, the application of genomics to clinical medicine is encouraging [START_REF] Wang | Genomics and drug response[END_REF] . Furthermore, the implementation of "-omics" technologies is expected to provide further important information concerning the activity of implicated mechanisms and pathways and the responses to the environment [START_REF] Voora | Clinical application of cardiovascular pharmacogenetics[END_REF][START_REF] Dorn | Decoding the cardiac message: The 2011 thomas w. Smith memorial lecture[END_REF] .

I have realised a pharmacological study dealing with the effect of clopidogrel on blood pressure levels, its interaction with calcium-channel blockers (CCB) and the effect of CYP2C19 gene polymorphisms on these interactions in a population of myocardial infarction patients coming from Coimbra University hospital in Portugal (n=2322). Clopidogrel use was

shown to be significantly associated with a lower diastolic blood pressure (beta = -1.47, P = 0.034). Following stratification of the population based on CCB use, the association of clopidogrel use with low diastolic blood pressure was observed only in the CCB users group.

Thus we have demonstrated that clopidogrel seems to improve the diastolic blood pressure lowering effect of CCB. In a population of healthy unrelated individuals from SFS (n=2155),

we have assessed the association of the rs4244285 polymorphism of CYP2C19 on blood pressure and significant associations were observed for both systolic and diastolic blood pressure (beta: -1.69, P = 0.008 and beta: -1.28, P = 0.003 respectively). This loss-of function polymorphism of CYP2C19 gene could be implicated in aldosterone regulation and thus in blood pressure levels and could be mediating the relation between clopidogrel and blood pressure. This work was presented in one original publication [START_REF] Stathopoulou | Newly identified synergy between clopidogrel and calcium-channel blockers for blood pressure regulation possibly involves cyp2c19 rs4244285[END_REF] (original publication 25), in 2 international conferences as oral presentations (conferences 18 and 20) and in one international conference as poster presentation (conference 19).

In another study, we have assessed whether interactions between CYP2C19 polymorphisms (loss-of-function (CYP2C19*2) and gain-of-function (CYP2C19*17)) and smoking are associated with the levels of platelet activation and CRP level in a post-PCI (percutaneous coronary intervention) population from the Centre Hospitalier Universitaire Timone in Marseille, after one month treatment with aspirin and either clopidogrel or prasugrel (n=1128). We have identified significant interactions between CYP2C19*2 polymorphism and CRP levels (P=0.005) and between metabolising status and smoking for CRP in the way that among smokers the highest CRP levels were observed in poor metabolisers. We have thus demonstrated that the interaction between smoking and CYP2C19 poor metabolising status is associated with higher CRP levels in post-PCI patients using thienopyridines. This work was presented in one original publication [START_REF] Shahabi | Genetic determined low response to thienopyridines is associated with higher systemic inflammation in smokers[END_REF] (original publication 21) and in two international conferences as poster presentation (conferences 9 and 14). In this study I have participated in data analysis and interpretation and reviewing of the manuscript.

Other projects

During the post-doctoral period I have participated also in other projects linked with the thematic of the IGE-PCV; U1122 unit.

In one nutritional study, we have assessed the associations of alcohol consumption and alcoholic-beverage preferences with dietary patterns in a sample of SFS (n=423 unrelated middle-aged males). We demonstrated a negative association between increased alcohol intakes and consumption of milk, yogurt and fresh/uncured cheese, sugar and confectionery, pastries and cookies, vegetables and fruit (P=0.002), and a positive relationship with cheese, meat and organs, pork-butcher's meat, and potatoes (P=0.011). Thus, we have verified that the associations of poorer dietary practice with alcohol consumption found in different populations around the world were observed also in a French male population. This work was published in one original publication [START_REF] Herbeth | Alcohol consumption, beverage preference, and diet in middle-aged men from the stanislas study[END_REF] (original publication 33).

My working experience in the research field of genetic epidemiology of CVD in the IGE-PCV; U1122 unit has resulted also in the drafting of several scientific reviews focused on this field. One review was focused on the challenges of GWAS in CVD [START_REF] Ndiaye | Cardiovascular diseases and genome-wide association studies[END_REF] (scientific review 9). A second review was related to an update on genetics of adverse cardiovascular events [START_REF] Visvikis-Siest | Common mutations and polymorphisms predicting adverse cardiovascular events: Current view[END_REF] (scientific review 6).

Participation in research contracts

As The aim of this thesis was the investigation of the genetic determinants of VEGF levels along with the pre-analytical and analytical variation factors. The identified polymorphisms that determine VEGF levels were tested against metabolic syndrome, blood lipids, adhesion and inflammation molecules, while associations with VEGF gene expression isoforms were also assessed.

The publications related to this thesis are: Following up the projects that were done during the post-doctoral period on VEGF, we aimed to continue our studies on the links between VEGF and CVD intermediate phenotypes, but also to further assess the role of VEGF on disease risk and its usefulness as a biomarker for personalised medicine.

In one study, we have assessed possible genetic connections between different angiogenesisrelated genes, including NOS3, CD14, MMP3, IL4R and IL4 with VEGF gene expression and plasma levels. The study was performed in 403 unrelated adults of SFS, where the variants rs2569190, rs2243250, rs2010963, rs3918226, rs1799983, rs1800779, rs3025058, rs1805015 and rs1801275 of the NOS3, CD14, MMP3, IL4R, and IL4 genes were selected to be genotyped on the basis of their functionality on the related genes. Furthermore, the 4 VEGFrelated SNPs were also assessed (rs6921438, rs10738760, rs6993770, rs4416670). Four mRNA isoforms of VEGF gene (VEGF-121, VEGF-145, VEGF-165, VEGF-189) were quantified in PBMCs of 65 individuals. We found that the SNP rs1799983 of NOS3 gene was associated with VEGF-145 mRNA levels (β = 0.17, P = 0.033). Five variants in NOS3, CD14, IL4R, MMP3 genes and one VEGF-related SNP were associated with VEGF plasma levels through epistatic interactions (P<0.05). Furthermore,; significant gene × environment interactions were identified for the rs1800779 SNP of NOS3 with levels of HDL, triglycerides and with obesity for VEGF plasma levels (P<0.05). This study demonstrated interactions between SNPs of angiogenesis-related genes with VEGF plasma levels and gene expression, especially for NOS3 gene, thus supporting the hypothesis that VEGF plays a central role in linking different angiogenetic mechanisms related to chronic diseases. These results were presented in one original publication [START_REF] Saleh | Angiogenesis related genes nos3, cd14, mmp3 and il4r are associated to vegf gene expression and circulating levels in healthy adults[END_REF] (original publication 19) and as a poster presentation in the 7th Santorini Conference Biologie Prospective (conference 12). This was the work of Abdelsalam Saleh thesis (doctoral student 5), which I co-supervised. For this project, I performed the study design, the analysis and interpretation of data and the drafting of the manuscript.

In the next step, we worked on a project that aimed to assess the links between VEGF and auto-immune thyroid diseases (AITD). A case control study was performed with 389 patients with AITD (297 with Hashimoto thyroiditis (HT) and 92 with Graves' disease (GD)) and 364

healthy controls recruited at the University Hospital Fattouma Bourguiba, Monastir, Tunisia.

The 4 VEGF-related SNPs were genotyped (rs6921438, rs10738760, rs6993770, rs4416670), and five VEGF mRNA expression isoforms were quantified in 71 patients and 86 controls (VEGF-121, VEGF-165, VEGF-145, VEGF-189 and VEGF165b) after 6 months of treatment. VEGF plasma levels were measured in 110 patients after 6 months of treatment.

We demonstrated that the rs10738760 was significantly associated with an increased risk of GD compared with HT (β = 0.54, P = 0.003) and controls (β = 0.46, p = 0.006). A significant association between rs6921438 and risk of GD compared with controls was also observed (β = 0.324, P = 0.05). The rs4416670 polymorphism was associated with an increased risk of hyperthyroidism in 6 months treated patients (β = 1.82, P = 0.049), independently of their initial diagnosis, while a significant positive association between the rs10738760 and plasma levels of FT3 (β = 0.716, P = 0.024) after 6 months treatment was observed. No significant differences were found between the groups for plasma VEGF levels. After 6 months of treatment VEGF-165 mRNA levels were higher in GD (P = 0.040), while the VEGF-189 was lower in GD (P = 10 -3 ), in HT (P = 10 -3 ) and in all individuals with AITD compared to controls (P = 10 -3 ). However, VEGF-165b was higher in HT compared to GD (p = 0.017).

VEGF-165b was more increased in individuals with hypothyroidism compared to euthyroid subjects (p = 0.043) after treatment. This study demonstrated for the first time associations between specific VEGF isoforms and different AITD after treatment and a common genetic link between VEGF and AITD is supported. These results were presented in 2 original publications [START_REF] Rancier | Pro-and anti-angiogenic vegf mrnas in autoimmune thyroid diseases[END_REF][START_REF] Zaaber | Plasma vegf-related polymorphisms are implied in autoimmune thyroid diseases[END_REF] (original publications 13 and 18). This was the work of Marc Rancier thesis (doctoral student 4) and of Jerome Chatelin thesis (doctoral student 2), which I co-supervise.

For this project, I performed the study design, the analysis and interpretation of data and the reviewing of the manuscript.

The next study on VEGF was focused on a new GWAS study aiming to identify novel polymorphisms that explain more of the individual variability of VEGF levels. A greater sample was used in this effort with 6 discovery cohorts (n= 13312) and in silico and de novo replication in n=2800 individuals. Six novel variants were identified that together with the 4 previously found (figure 5) explain 52% of the variability of VEGF levels. The novels variants are linked with known and novel pathways associated with VEGF actions. This is the biggest GWAS study on VEGF levels today. These results are presented in one original publication [START_REF] Choi | Six novel loci associated with circulating vegf levels identified by a meta-analysis of genome-wide association studies[END_REF] (original publications 17) and in one conference as a poster presentation (conference 15). In this work I have performed the design of the replication, the analysis of SFS data, the interpretation of the results and the reviewing of the manuscript.

In the following study, we aimed to assess the genetic effect of the 4 VEGF-related polymorphisms (rs6921438, rs10738760, rs6993770, rs4416670) on the risk of depression. A case-control population of 437 patients diagnosed with depressive disorders and 477 healthy controls was recruited in the Mental Health Centre of Extremadura University Hospital, Badajoz, Spain. We have identified a direct effect of rs4416670 on depression risk (OR: 1.60, P=0.010). This study demonstrated genetic links between VEGF and depression for the first time. As we have previously associated the rs4416670 with gene expression of L-selectin [START_REF] Azimi-Nezhad | Associations of vascular endothelial growth factor (vegf) with adhesion and inflammation molecules in a healthy population[END_REF] , we hypothesised that this SNP association with depression could be mediated by VEGF and L-selectin. These results are presented in an original publication [START_REF] Xie | Vegf-related polymorphisms identified by gwas and risk for major depression[END_REF] (original publication 11).

This work was part of the thesis of Ting Xie (doctoral student 3), which I co-supervised. For this work, I have performed the study design, the interpretation of the results and the drafting of the manuscript.

In a collaborative study, we have assessed the effect of the 4 VEGF-related polymorphisms on the risk of metabolic syndrome in an Iranian population (n=336) (rs6921438, rs10738760, rs6993770, rs4416670). We have demonstrated that rs6993770 was associated with an increased risk for metabolic syndrome (P = 0.007). This work was published in one original publication [START_REF] Azimi-Nezhad | The relationship between vascular endothelial growth factor cis-and trans-acting genetic variants and metabolic syndrome[END_REF] The disturbances of lipid homeostasis in peripheral tissues, named dyslipidemias, have been clearly shown to be significant risk factors for atherosclerosis, the underlying cause of CVD [START_REF] Kopin | Dyslipidemia[END_REF] . The apolipoprotein E (ApoE) plays an important role in lipid homeostasis by acting as a ligand for lipoprotein receptors. It is involved in particular in the hepatic clearance of lipoproteins rich in triglycerides [START_REF] Upadhyay | Emerging risk biomarkers in cardiovascular diseases and disorders[END_REF] . The APOE gene is located on chromosome 19q13.1, a locus related to hyperlipidemia as indicated by GWAS 96 . The APOE gene has a wellcharacterised three-allele polymorphism (ε2, ε3 and ε4) that determines the variability of an individual's lipid profile. The ε3 allele is the most common allele and is considered the wildtype allele. The ε4 variant is linked to high levels of plasma lipids. In addition, the common polymorphism of the APOE gene is related to type III hyperlipidemia (type III HLP, OMIM 107741), a familial dyslipidemia characterised by the accumulation of TG-rich lipoprotein residues, leading to an increased risk of atherosclerosis. [START_REF] Mahley | Type iii hyperlipoproteinemia (dysbetalipoproteinemia): The role of apolipoprotein e in normal and abnormal lipoprotein metabolism[END_REF] . Carriers of the APOE ε2 / 2 genotype show the highest probability of having a type III HLP. On the contrary, as indicated previously [START_REF] Salah | Combined effects of lipoprotein lipase and apolipoprotein e polymorphisms on lipid and lipoprotein levels in the stanislas cohort[END_REF][START_REF] Pallaud | Genetic influences on lipid metabolism trait variability within the stanislas cohort[END_REF] , we found significant associations of the APOE ε2 allele with reduced levels of triglycerides, cholesterol and LDL and increased levels of ApoE in adults LSR represents a potential link between dyslipidemia and atherosclerosis. Although the absence of both LSR alleles is lethal at the embryonic stage, studies in the LSR +/heterozygous mice revealed that reduced LSR levels led to high postprandial lipemia, moderate hypercholesterolemia, and hypertriglyceridemia. 108 . LSR +/-heterozygous mice placed on a cholesterol-rich "Western" diet show an accumulation of atherogenic lipoproteins such as LDL and triglycerides-rich lipoproteins, which is accompanied by an increase in lipid deposition in the aorta 108 . Hepatic LSR knockout results in both hypertriglyceridemia and hypercholesterolemia in mice, confirming that LSR contributes significantly to the regulation of lipid status 110 . Cell culture studies (fibroblasts, hepatocytes) have demonstrated the ability of lipid particles containing ApoE to bind to LSR, leading to their internalization and degradation in cells 111,112 . In addition, the binding affinity seems different according to the APOE variants: the very low density lipoproteins (VLDL) of a patient ε2 / 2 with HLP type III are unable to bind to the LSR 112 . The low affinity of LSR for VLDL of a patient ε2 / 2 could contribute to a decrease in hepatic clearance of triglycerides-rich lipoproteins, leading to an increase in plasma triglycerides observed in this particular class of hyperlipidemia.

Moreover, dyslipidemic LSR+/-mice display age-dependent modified brain cholesterol trafficking, increased levels of membrane cholesterol 113 along with increased susceptibility to amyloid stress 114 . LSR is important for blood brain barrier (BBB) integrity 115,116 , which may explain why hypertriglyceridemia has been associated with BBB impairment 117,118 often observed in AD. Taken together, these results suggest that LSR pathway may represent an important molecular link between dyslipidemias and risk for cognitive decline and AD.

The LSR gene that encodes LSR is also located on chromosome 19q13.1 just upstream of APOE. Little information is currently available on the role of LSR polymorphisms in the regulation of lipid levels. SNP rs916147 (A / G), located in a splice junction between intron 5 and exon 6, was associated with lipid levels in a small population of obese adults 119 . In addition, this SNP is a quantitative expression locus (eQTL) for LSR and can alter the expression of LSR in different tissues (subcutaneous and visceral adipose tissue, tibial artery, coronary artery and skeletal muscle), thereby indicating its potential functionality (www.gtexportal.org). The SNP rs34259399 is located on exon 6 and preliminary unpublished data suggest that it is involved in the regulation of LSR expression and activity.

The role of these two functional SNPs on AD risk is unknown. We have thus hypothesized that APOE variants could interact with LSR polymorphisms to modify the effect of APOE on blood lipid regulation and AD risk.

In the first study, we have used a discovery population of healthy adults and children coming from the SFS (n=432 unrelated adults and n=328 children (<18 years old)) and a replication population of n = 1744 unrelated adults to test the association of the direct and epistatic effects of LSR and APOE variants on blood lipids levels (cholesterol, triglycerides, HDL, LDL, ApoB and ApoE). We have found significant epistatic interactions between the LSR rs916147 and APOE ε2 allele in adults for cholesterol (b=0.114, P=0.810 × 10 -8 ), ApoE (b=0.125, P=0.625 × 10 -3 ) and LDL levels (b=0.059, P=0.120 × 10 -3 ). These results replicated for cholesterol and ApoE. The interaction of the LSR rs916147 with APOE ε2 reversed the direct effect of APOE ε2 on cholesterol, ApoE and LDL levels. We could therefore suggest that for APOE ε2 subjects, the interaction with the rs916147 could lead to an increased risk for APOE ε2/2 subjects to develop type III HLP. These results were presented in one original publication 120 (original publication 6) and were presented as a poster presentations in 2 international conferences (conference 1 and 8).

In a second publication, we investigated the effects of the LSR and APOE polymorphisms on AD risk. A case-control population of 142 patients diagnosed with AD and a control group of 63 healthy volunteers of European origin was used. We identified significant epistatic interactions between the 2 functional LSR SNPs and APOE ε4 allele for AD: rs34259399 × APOE ε4 (beta= -0.95, P=0.00002) and rs916147 × APOE ε4 (beta=-0.83, P=0.0068). In more details, all non-ε4 allele carriers had increased risk of AD when combined with the with the LSR polymorphisms. These results were published in one original publication 121 (original publication 3).

These two studies were parts of the thesis of Ting Xie (doctoral student 3), whom I cosupervised. My involvement included the study conception and design, data analyses and interpretation and drafting of the manuscripts.

Project 3: Triggering Receptors Expressed on Myeloid cells (TREM) and inflammation

Triggering Receptor Expressed on Myeloid cells-1 (TREM-1) is a member of a recently discovered immunoglobulin family 122,123 . It is implicated in an increase of the inflammatory responses of neutrophils and monocytes 124 and is related to the production of proinflammatory cytokines (TNF-α and IL1b) 125 . TREM-1 apart from its role during the pathophysiology of septic shock 122 , it is also involved in other acute and chronic inflammatory diseases of non-infectious etiology, such as rheumatoid arthritis 126 , atherosclerosis 127 , acute myocardial infarction 128 and critical limb ischemia 129 . The mechanisms that explain this action are not yet clearly described. Adhesion molecules like selectins have an important role during this process [130][131][132] . In the one study, we have investigated the genetic links between variants within or near the TREM-1 gene and soluble TREM-1 (sTREM-1 and TREM-1sv) serum levels and with the expression levels of two TREM-1 splicing isoforms in PBMCs, the membrane form (mbTREM-1) and the soluble form (TREM-1sv). Further, we have tested the associations between the significant these SNPs and levels of soluble E-selectin, L-selectin and P-selectin, which are involved in the early stages of atherosclerotic processes. A discovery cohort with 351 individuals from SFS and an independent replication population of 80 subjects was used. We have included 10

SNPs in the analysis from which the rs2234246 was associated with increased sTREM-1 levels in the discovery population (b = 0.3, P = 0.003), and with increased levels of mRNA of mbTREM-1 isoform (b= 0.49, P = 0.007). The same SNP was associated with increased soluble L-selectin levels (b = 0.05, P = 0.011). The associations of the SNP with sTREM-1 and L-selectin levels were confirmed in the replication population (b = 0.13, P = 0.0007 and b= 0.03, P = 0.018 respectively). In this study, we identified one polymorphism affecting sTREM-1 levels and mbTREM-1 isoform in PBMCs, which is also associated with L-selectin levels suggesting that TREM-1 could act in the trans-epithelial migration process of leukocytes and more specifically of monocytes through expression of L-selectin. This work was presented in one publication 133 (original publication 10) and was presented in one international conference as a poster presentation (conference 3).

Triggering Receptor Expressed on Myeloid cells-2 (TREM-2) is expressed on the cell surface of macrophages and dendritic cells derived from monocytes, as well as in microglia and osteoclasts. TREM-2 plays a role in a series of tyrosine phosphorylation reactions that regulate various inflammatory responses 125,134 . It exerts anti-inflammatory properties during the immune response 135,136 , including stimulation of phagocytosis and suppression of cytokine production like the TNF-α 137, 138 and it is thus linked with atherosclerosis. We have therefore assessed the effect of TREM-2 polymorphisms on TNF-α levels and intima media thickness of the femoral artery in 809 children and adults from the SFS and in 916 unrelated adults of an independent replication cohort. We have demonstrated significant associations between the SNP rs6918289 with plasma levels of TNF-α in discovery and replication populations (b=0.13, P=0.0026, and b=0.20, P=0.023, respectively), and with femoral artery thickness in the discovery cohort (b= 0.02, P=0.026). Thus, this SNP could be a risk factor for inflammatory diseases and atherosclerosis. These results are presented in one publication 139 (original publication 4) and in 2 international conferences as poster presentations (conference 4 and 6). This work was part of the thesis of Vesna Gorenjak, which I co-supervise (doctoral student 1).

For both these projects I participated in the data interpretation and the reviewing of the manuscripts.

Project 4: Telomere length as a biomarker for personalised medicine

Continuing the research activities on TL genetics and its use as a biomarker in personalised medicine, we have designed different projects, which make part of the thesis of Vesna Gorenjak, which I co-supervise (doctoral student 1). A scientific review underlining the importance of telomere length in personalised medicine has been published with my participation in the data collection and drafting of the manuscript 140 (scientific review 2).

In a candidate gene study, we are assessing the effect of previously identified genetic determinants of LTL and of novel candidate genes including the 10 VEGF-related SNPs with muscle TL (MTL) in the case-control population TELARTA (ANR Programme de Recherche Translationnelle en Santé) (n= 451). MTL is reflecting the TL at the time of birth, as muscle cells do not undergo cell divisions. Thus, the study will identify whether the candidate genes assessed affect the TL at birth or they are implicated in TL attrition. Statistical analysis is ongoing.

Other projects on TL include new GWAS and EWAS efforts including novel bioinformatics approaches in collaboration with international consortia such as the CHARGE consortium Aging Working Group, which are under development. All these projects and the preliminary results were presented as poster presentation in the 9th Santorini Conference in 2018 (conference 2).

Project 5: Novel genetic determinants of inflammation molecules

Inflammation is now considered a basic biological process implicated in atherosclerosis as its role is central to all stages of atherosclerosis. As a result, inflammation biomarkers have been extensively studied in relation to CVD traits, especially CRP, which has also been included in recent risk prediction algorithms 141 . Inflammation represents one the basis axes of research of the IGE-PCV; U1122 unit.

In one recent study, we have investigated the associations between IL6R polymorphisms with CRP and lipids levels in a healthy population. We have used 368 unrelated adults from SFS as discovery cohort and 995 adults as an independent replication population. The variants rs4845625 and rs4537545 of IL6R were selected based on their contradictory role on IL6 and blood lipids levels. Both polymorphisms were significantly associated with CRP levels (rs4845625 beta =0.182, P=0.011 and for s4537545 beta =-0.189, P=0.009). The rs4845625 was associated with LDL in both populations (beta =0.052, P=0.007, and beta =0.024, P=0.035). Haplotypes analyses confirmed these results. In silico analysis revealed that these SNPs are linked with genes involved in the regulation of the JAK-STAT pathway cascade that affects lipid metabolism. This work was presented in one original publication 142 (original publication 9)

Aiming to increase the impact of our studies in the inflammation field, the unit has developed collaboration with the CHARGE (Cohorts for Heart and Aging Research in Genomic Epidemiology) consortium Inflammation Working Group (CIWG), which I lead as a principal investigator for SFS.

I have participated in the data interpretation and the drafting of the manuscript (member of the writing group) of the most recent and largest GWAS on CRP 143 (original publication 1). The study included 88 cohorts with 204402 European individuals. We have identified 58 distinct variants significantly associated with levels of CRP, including 16 previously identified CRP loci. The associations are explained by 55 genes that are involved in the biology of the immune system and liver. Mendelian randomisation analyses reported causal associations between genetically increased CRP and a protective effect on schizophrenia and increased risk of bipolar disorders.

Two more projects on GWAS meta-analysis for IL6 and TNF-α levels are being completed within the CIWG. For these projects; I have performed the data preparation and analysis for SFS and we are currently in the phase of reviewing the related manuscripts.

cohorts were analysed including SFS. The first study was a GWAS meta-analysis of 339224 individuals from 125 studies that resulted in identification of 56 novel BMI-associated loci, implicated in pathways related to synaptic function, glutamate signalling, insulin secretion/action, energy metabolism, lipid biology and adipogenesis 166 . The second study was GWAS meta-analysis of 224459 individuals that identified 33 novel waist/hip circumference associated variants and 19 novel SNPs associated with waist and hip circumference measures involved in pathways of adipogenesis, angiogenesis, transcriptional regulation, and insulin resistance. In these studies I was the analyst of the SFS study data and reviewed the related manuscripts. The third study assessed the genome-wide burden of runs of homozygosity on 16 complex traits in 354224 subjects from 102 studies and demonstrated the existence of directional dominance on four complex traits (stature, lung function, cognitive ability and educational attainment). In this study my contribution was on databases preparation and sharing.

Furthermore, I have participated as the analyst of the SFS in multiple projects of the Global Burden of Metabolic Risk Group Factors of Chronic Disease Collaborating Group (NCD Risk Factor Collaboration -NCD-RisC). I have performed descriptive analysis on height, weight, BMI, blood pressure, blood lipids levels and insulin levels in SFS as indicated by the analysis plan of each study shared within the consortium. The 6 produced publications are presenting the trends of these traits in very large samples across the globe 169-174 (original publications 5, 8, 12, 14, 15, 16).

Moreover, I have contributed in the data collection and drafting of 3 scientific reviews: one on personalised medicine major achievements, one on pharmacogenomic challenges in CVD and one on genetics beyond GWAS, based on the experience and expertise of the IGE-PCV;

U1122 Unit on the field of genetics of CVD [175][176][177] (scientific reviews 1, 3 and 5). Two of them are parts of the thesis of Jerome Chatelin and Vesna Gorenjak, which I co-supervise (doctoral students 2 and 1).  Inflammation project on childhood, which is an ongoing project where I am the leader and principal investigator within the CHARGE consortium (details on Project "Novel genetic determinants of inflammation molecules" above).

Participation in international consortia

Global Burden of Metabolic

Supervision of students and participation in evaluation committees

Since my nomination as a research engineer in Inserm in 2015, I have co-supervised 3 doctoral students having an official authorization (ACT) from the doctoral school BioSE.

These were: This thesis is aiming to study the effect of genetic variants related to VEGF and its expression isoforms with the risk factors and the risk of developing chronic diseases including CVD, osteoporosis, and thyroid diseases in order to demonstrate their clinical utility in personalised medicine.

The so far accepted publications related with this thesis are: This thesis aims to study the telomere length and the VEGF as biomarkers for personalised medicine. Modern approaches will be used like mendelian randomisation and the study of epigenetic modifications and will be combined with classical approaches of GWAS and candidate genes in order to clarify the determinants of telomere length, VEGF and associated molecules and their association with intermediate phenotypes of CVD.

The so far accepted publications related with this thesis are: I am also member of the emergency standby personnel for the security of biological material stored in the freezers of the BRC IGE-PCV. I have the responsibility of a mobile phone that informs of the slightest problem with the temperature of the freezers and I must intervene on the spot in case of need on a plan of 24h / 24h and 7 days / 7 days.

Administrator of the "Omics_BioBook" platform

As mentioned before, this platform is a Biostatistical / Bioinformatics platform, developed in with the aim of linking clinical data, biological data, "-omics" data and sample data for BRC IGE-PCV populations. This interface will facilitate the search for populations according to project-specific selection criteria and the creation of the databases that will be used for statistical analysis. An SFTP server has also be installed to ensure the safe transfer of big data between the unit and our collaborators and consortia.

Administrative activities

I am a member of the laboratory council of the unit IGE-PCV; U1122. I participate in the annual meetings that have as aim to:

1. Give an opinion on the main scientific policy decisions of the laboratory, changes in its structure, human resources management, research contract policy, technology transfer policy, training program, budget and finances 2. To make reflections on the main strategic directions of the unit on the medium and long term.

Introduction

My scientific interest and my research experience on the field of VEGF as a biomarker useful in personalised medicine has led to the development of this research project that will be developed during the following years within the frames of the IGE-PCV ; U1122 unit and the VEGF Consortium.

According to the World Health Organization (WHO), non-communicable diseases are responsible for >71% of all deaths worldwide. Among them, CVD (like heart attacks and stroke), cancers, chronic respiratory diseases (such as chronic obstructive pulmonary disease and asthma) and diabetes account for over 80% of all premature non-communicable diseases deaths 178 . CVD, including all the diseases of the heart and circulation, represent the primary cause of death worldwide 179 . CVD have high prevalence and economic impact: overall cost of CVD to the EU economy is €210 billion a year 180 . They require tailored management strategies, thus personalised early risk prediction is an urgent need for European citizens and healthcare systems.

Angiogenesis is a complex process promoting the development and growth of new vascular structures in contact with pre-existing vasculature. As mentioned before, it plays a significant role in physiological conditions but it is also involved in a wide-range of pathological conditions including vascular diseases and cancer. Induction of angiogenesis is regulated by a variety of molecules such as the VEGF family, TNF-α, interleukins, chemokines, and growth factors 181 . However, VEGF is the most potent factor associated with angiogenesis and is considered a key regulatory molecule for both physiological and pathological angiogenesis 19 .

Atherosclerosis is a chronic inflammatory condition characterized by lipid-containing inflammatory lesions of large and medium-sized arteries. These lesions cause a focal thickening of the intima resulting in local hypoxia, which is enhanced by the inflammatory cell content of the lesions and the produced inflammatory factors 182 . Hypoxia activates a metabolic pathway mediated mostly by the transcriptional factor HIF-1 alpha (hypoxiainducible factor 1-alpha), which up-regulates the expression of various genes including nitrous-oxide synthetase (NOS) and VEGF. The effect of VEGF actions in the vasculature can have a positive and a negative impact. The production of NO and prostacyclin (PGI2) from VEGF activation has an important role in maintenance of endothelium integrity, in the lowering of vascular tonus and in the inhibition of platelet aggregation and thrombosis. It also induces collateral angiogenesis and thus restores blood circulation in infarcted areas 27 .

However, the neovessels formed in the atherogenic plaques express adhesion molecules such as VCAM-1, ICAM-1 and E-selectin, which allow the leukocyte infiltration in atherosclerotic plaque (figure 6). The recruited leukocytes produce inflammation molecules and various metalloproteinases (MMPs), which contribute to the instability of the plaque. Furthermore, VEGF cell activation leads to increased production of inflammation factors such as TNFa and

IL1b. The increased vascular permeability and the VEGF-induced attraction result in increased macrophages infiltration. These inflammation mechanisms further enhance the VEGF expression and persistence of angiogenic stimuli, resulting in pathological angiogenesis. Moreover, the vascular permeability causes the extravasation of erythrocytes, which in combination with the fact that newly developed vessels are dysmorphic and immature, results in intraplaque hemorrhage 22,23,27 . These complex procedures finally result in plaque instability which is the major cause of most severe clinical events, as plaque rupture can occur and cause sudden thrombosis of the artery 182 . with an elevated risk of side-effects such as hypotension, while gene therapy was better tolerated 183 . Moreover, as mentioned before, the anti-VEGF factors used in cancer lead to a range of cardiovascular side-effects including hypertension, cardiac ischemia, arterial thromboembolic events (e.g. cerebral and myocardial infarction) and left ventricular dysfunction 27 . Furthermore, it seems that the relationship between VEGF and CVD is complicated also due to pleiotropic effects of VEGF. Apart from its angiogenic functions, VEGF exerts a variety of actions in other organs, through its complicated signaling pathways in ECs and other cells, which explains its implication in a wide range of conditions such as inflammation, vascular diseases, hypertension, cardiac, liver and kidney function 184 .

Angiogenesis and VEGF are considered as a common pathway linking different noncommunicable diseases that also represent a huge burden for public health.

Neurodegenerative diseases such as AD and osteoporosis are common non-communicable diseases with a high burden for the society and healthcare systems. Neurodegenerative diseases are major challenge for public health, are mostly incurable and have a high prevalence and cost in Europe: dementias alone are estimated to affect 7 million people and their cost is €130 billion a year 185 . Osteoporosis is also a disease with great prevalence and importance in public health. In Europe 200 million women worldwide are affected.

Nevertheless, it is a disease poorly identified and treated (80% of individuals at risk are not identified or treated) 186 .

VEGF seems to play a complex role in the central nervous system, and in dementia and AD development. VEGF is produced and secreted by astrocytes and through its receptors it activates pathways of cell survival, angiogenesis, and vascular cell permeability in the central nervous system 187 . Increased VEGF production after an ischemic stroke leads to cerebral angiogenesis and increased BBB permeability 188 . In brain tissue of AD patients, reduced VEGF gene expression has been detected, an indication of pathological angiogenesis 189,190 .

Increased serum VEGF levels in AD patients with cerebral microbleeds 191 , while lower VEGF levels have been observed in AD patients in another study 192 . Recently, VEGF levels in cerebrospinal fluid of AD patients show no differences compared to healthy controls 193 .

However, other studies demonstrate increased intrathecal levels of VEGF in AD patients 194 , and higher VEGF expression in microvessels isolated from AD patients brain tissues compared to controls 195 , thus indicating a complex role of VEGF in AD pathophysiology.

Genetic studies have also given controversial results concerning the role of VEGF polymorphisms and risk for AD [196][197][198] .

Concerning the link between VEGF and osteoporosis, this is also well documented.

Vasculature is important for osteogenesis 199 , the development of bones and for bone remodeling and fracture healing, as it is important for the transfer of precursors of osteoclasts They have originally identified the high heritable component of VEGF in SFS, which is more that 60% [START_REF] Berrahmoune | Heritability for plasma vegf concentration in the stanislas family study[END_REF] . This finding has been also verified by two partners of the VEGF Consortium 206,207 . This is an extreme genetic component and this makes VEGF unique among other biomarkers of chronic diseases. Up to half of this heritability is explained by 4 very common variations (frequency between 35-50%) identified by a GWAS, where the unit participated with SFS population [START_REF] Debette | Identification of cis-and trans-acting genetic variants explaining up to half the variation in circulating vascular endothelial growth factor levels[END_REF] , while 6 more variants have been identified in a more recent VEGF GWAS [START_REF] Choi | Six novel loci associated with circulating vegf levels identified by a meta-analysis of genome-wide association studies[END_REF] . This is also an exception in the field of genetic studies of complex diseases, as for most markers, the variability explained by identified variants does not reach levels higher than 5-10%. It is therefore a biomarker ideal for population stratification. Significant associations have been identified between these polymorphisms and risk factors of chronic diseases (cholesterol levels, blood pressure and inflammation molecules) [START_REF] Stathopoulou | A common variant highly associated with plasma vegfa levels also contributes to the variation of both ldlc and hdl-c[END_REF][START_REF] Azimi-Nezhad | Associations of vascular endothelial growth factor (vegf) with adhesion and inflammation molecules in a healthy population[END_REF] , with angiogenesis molecules [START_REF] Saleh | Angiogenesis related genes nos3, cd14, mmp3 and il4r are associated to vegf gene expression and circulating levels in healthy adults[END_REF] , with risk for depression [START_REF] Xie | Vegf-related polymorphisms identified by gwas and risk for major depression[END_REF] and with thyroid hormones levels in patients with autoimmune thyroid diseases [START_REF] Rancier | Pro-and anti-angiogenic vegf mrnas in autoimmune thyroid diseases[END_REF][START_REF] Zaaber | Plasma vegf-related polymorphisms are implied in autoimmune thyroid diseases[END_REF] . Furthermore, as discussed before, VEGF is implicated in the pathophysiology of CVD and other chronic diseases, while the existence a large number of transcription isoforms (and related protein products) from the VEGF gene can allow the study of VEGF specificity for CVD and other diseases. These arguments demonstrate that VEGF represents an ideal biomarker for the application of "-omics" methodologies in personalised medicine of chronic diseases. Therefore, the in-depth investigation of VEGF and its clinical use in chronic diseases risk prediction can have an important impact in public health.

In line with this and using my past experience and expertise on VEGF and public health research field, the purpose of the proposed project is the development of VEGF-based CVD risk prediction model based on the validity and utility of its molecular determinants as CVD biomarkers. The most recent "-omics" methodologies will be applied for the identification of molecular determinants of VEGF that explain the largest part of its inter-individual variability.

Objectives

The general aim of the project is to develop risk prediction algorithms for the stratified risk prediction of CVD and other chronic diseases based on VEGF "-omics" determinants. Big data from "-omics" determinants (genetics, transcriptomics and epigenetic) of VEGF and VEGF-related molecules will be combined with environmental factors and epistatic interactions with other molecules involved in pathophysiological processes common to the assessed diseases (angiogenesis, inflammation, lipid metabolism). The project's specific objectives are:

1. The identification of VEGF "-omics" determinants and their interactions with environmental/lifestyle factors in health through robust and valid multi-disciplinary "omics" assays (including genetics, epigenetics, and transcriptomics) in large familial, longitudinal populations of healthy individuals of European ancestry.

2. The identification of VEGF "-omics" determinants that affect CVD, AD and osteoporosis risk factors, as well as their interactions with environmental/lifestyle factors in healthy populations.

3. The identification of VEGF "-omics" determinants that affect CVD, AD and osteoporosis risk in case-controls and prospective studies. This study will allow to test the specificity of different "-omics" determinants on different diseases.

4. The development of VEGF "-omics"-based computational risk prediction models and algorithms. Their predictive value will be assessed in prospective studies aiming to identify at-risk groups among individuals throughout their lives. Their utility and stratification efficacy will be compared to conventional risk prediction algorithms, taking into account both age and sex.

The project will use the resources available in the IGE-PCV; U1122 unit and the VEGF Consortium that is coordinated by the unit. Resources in population include recognised cohorts (SFS, Framingham Heart Study (FHS); Cilento study, LLs, ORIGIN), case-controls populations, large populations from biobanks (BRC IGE-PCV, BBMRI-ERIC a pan European infrastructure that regroups more than 250 biobanks and UK Biobank) and collaborations with other consortia such as CHARGE, METASTROKE and MEGASTROKE for which the desired "-omics" data are available. The "Omics_BioBook" platform will be used for data storage and analyses.

Implementation of the project

The project will be based on 4 axes, one for each specific objective.

Axe 1: Identification of VEGF molecular determinants in health

The main objective of axe 1 is to identify the genetic, transcriptomic and epigenetic determinants of VEGF biology in health with the use of robust "-omics" assays validated in large familial, longitudinal cohorts of healthy individuals.

Genetic determinants of VEGF levels

The results of the recent GWAS study on VEGF levels will be expanded with the addition of new populations that are available through CHARGE consortium and the ORIGIN Study and new GWAS meta-analyses. The aim is to gather more than 20000 individuals thus increasing the power of the study and the potential for identification of novel SNPs that will increase the percentage of variability of VEGF levels explained. A list of GWAS-level significant independent SNPs will be developed, while SNPs reaching a level of significance of 0.05 by false discovery rate correction (less strict criterion compared to classical Bonferroni correction applied in GWAS) can also be added in the list of candidate genetic determinants of VEGF. Novel discoveries will also be included by gene × environment interactions studies, where environmental factors known to affect the risk of CVD, AD and osteoporosis will be selected: BMI, levels of blood lipids, inflammation molecules, blood pressure etc. Large scale replications will be performed in UK Biobank. A genetic score of all the independent markers will be developed.

Transcriptomic determinants of VEGF levels

RNA sequencing through TrueSeq (Illumina) and sequencing on HiSeq2500 instruments (75 bp paired end reads) has been performed for 220 individuals from 70 families of SFS. These data and other RNA sequencing and qPCR methods data available in the healthy populations participating in the VEGF Consortium will be combined so as to quantify the expression of pro-and anti-angiogenic isoforms of VEGF gene in PBMCs. Following their quantification, association analysis and meta-analyses will be performed in order to identify the isoforms that affect more the levels of VEGF.

Epigenetics determinants of VEGF levels

DNA methylation profiling with the Infinium Methylation450K beadchip (Illumina) that can interrogate 450,000 CpG sites and quantify DNA methylation levels is available for 220 individuals from SFS and the other healthy populations participating in the VEGF Consortium. EWAS meta-analyses will be performed to identify methylation probes that are significantly associated with VEGF levels.

Axe 2: Identification of VEGF molecular determinants associated with chronic diseases risk factors

The main objective of the axe 2 is to establish and quantify the effect of VEGF "-omics" determinants on common to the assessed chronic diseases risk factors and their interactions with environmental/lifestyle factors. These disease risk factors include blood lipids, blood pressure, glucose metabolism, obesity and inflammation traits such as systolic blood pressure, diastolic blood pressure, mean arterial blood pressure, CRP, haptoglobin, IL-6, TNF-α, fasting blood glucose, fasting insulin levels, BMI, waist circumference, waist/hip ratio, HDL, LDL, ApoE, triglycerides, lipid accumulation product index (LAP) and visceral adiposity index (VAI). Ten-year risk of a first fatal atherosclerotic event, such as heart attack, stroke, aneurysm of the aorta, or other calculated by SCORE system will also be used as a phenotype of interest 208,209 . These phenotypes are available in SFS, LLs, Cilento, FHS but also in many studies of the BRC IGE-PCV, of CHARGE consortium and of UK Biobank, gathering a powerful sample size. Direct, gene × gene and gene × environment interactions will be tested for the identified "-omics" determinants of the axe 1. This axe will identify specific VEGF "omics" determinants that affect specific pathways and diseases risk factors.

Axe 3: Identification of VEGF molecular determinants associated with chronic diseases risk

The main objective of the axe 3 is to identify the disease-specific VEGF "-omics" determinants that could be used for disease risk prediction. Case-controls populations with CVD (coronary artery disease and stroke), AD and osteoporosis will be used. These populations are available in the BRC IGE-PCV, The VEGF, CHARGE, METASTROKE and MEGASTROKE consortia, the BBMRI-ERIC and the UK biobank. In these populations, the identified from axe 1 and axe 2 VEGF "-omics" determinants will be tested for associations with increased risk for the assessed diseases. We expect to identify specific "-omics" determinants for each disease and lead to the selection of the strongest biomarkers to be added in the risk prediction models of the next axe.

Axe 4: Development and assessment of predictive value of personalised risk prediction algorithms based on VEGF "-omics" determinants

The main objective of the axe 4 is to develop and demonstrate the independent and additive predictive value of specific to CVD, AD and osteoporosis VEGF "-omics" determinants, identified in the previous axes, in computational prediction algorithms for the assessment of disease risk. The predictive value will be assessed in large prospective cohorts of European ancestry with the aim to identify at-risk groups among individuals throughout their lives. The incremental predictive utility and stratification efficacy of the new algorithms will be compared to conventional risk prediction algorithms such as the SCORE system 208 , the Framingham General Cardiovascular Risk Profile (FGCVDRP) 209 for CVD and the FRAX WHO Fracture Risk Assessment Tool for osteoporosis 210 .

The steps that will be followed for the development and the testing of the new personalised "omics" algorithms are 211-213 :

1. Population and end-points: prospective cohorts like FHS, LLs, ORIGIN, and others available in CHARGE consortium and the BBMRI-ERIC infrastructure. End points will include severe events of CVD, fractures and AD diagnosis.

2. Predictors: Defined in axe 2 and 3.

3. Model estimation: Sex-specific Cox proportional hazards regression is most commonly used for time-to-event data, such as long-term mortality (following testing of proportionality of hazards). The estimation of the regression coefficients for Cox regression models will be performed with maximum likelihood methods. To limit overfitting of models to the available data, techniques such as statistical shrinkage techniques, penalised maximum likelihood estimation, and the least absolute shrinkage and selection operator will be used.

4. Assessment of the model performance and validity: internal validation (reproducibility) will be performed using methods of cross-validation. External validation (transportability between the two cohorts) will also be performed. Calibration of the models (agreement between observed outcomes and predicted) will be assessed with goodness of fit methods and predicted-to-observed ratios. Discrimination is commonly quantified with a concordance (c) statistic. For a binary endpoint, c is identical to the area under the receiver operating characteristic (ROC) curve, which plots the sensitivity (true-positive rate) against specificity (false-positive rate) for consecutive cut-offs for the predicted risk. For a time-to-event endpoint, such as survival, the calculation of c may be affected by the amount of incomplete follow-up (censoring). We note that the c statistic is insensitive to systematic errors in calibration, and considers the rather artificial situation of classification in a pair of patients with and without the endpoint. The clinical usefulness will be assessed by the net benefit. This measure is consistent with the use of an optimal decision threshold to classify patients. The relative weight of harms and benefits is used to define the threshold, and is used to calculate a weighted sum of trueminus false-positive classifications. For the comparison of additive value and the performance between the novel and the conventional algorithms, methods such as continuous net reclassification improvement (NRI) and integrated discrimination improvement (IDI) will be used.

  School of Athens, National and Kapodistrian University of Athens, Harokopio University of Athens, Greece. Fund by Pharmaserve Lilly S.A.C.I.Research contracts as Principal Investigator (PI)2018AAP-BMS interne "Projets incitatifs -Crédits CS-UL 2018", Project :

2 .

 2 Azimi-Nezhad M, Mirhafez SR, Stathopoulou MG, Murray H, Ndiaye NC, Bahrami A, Varasteh A, Avan A, Bonnefond A, Rancier M, Mehrad-Majd H, Herbeth B, Lamont J, Fitzgerald P, Ferns GA, Visvikis-Siest S, Ghayour-Mobarhan M. The Relationship Between Vascular Endothelial Growth Factor Cis-and Trans-Acting Genetic Variants and Metabolic Syndrome. Am J Med Sci. 2018 Jun;355(6):559-565.3. Xie T*, Akbar S*, Stathopoulou MG*, Oster T, Masson C, Yen FT, Visvikis-Siest S. Epistatic interaction of apolipoprotein E and lipolysis-stimulated lipoprotein receptor genetic variants is associated with Alzheimer's disease. Neurobiol Aging. 2018 May 3. pii: S0197-4580(18)30147-7. * equal first authors 4. Gorenjak V*, Aldasoro Arguinano AA*, Dadé S, Stathopoulou MG, Vance DR, Masson C, Visvikis-Siest S. The polymorphism rs6918289 located in the downstream region of the TREM2 gene is associated with TNF-α levels and IMT-F. Sci Rep. 2018 May 8;8(1):7160. * equal first authors Supervision of students and participation in evaluation committees Co-direction (50%) of doctoral students with official autorisation (Autorisation à Codiriger une Thèse, ACT) 1. Vesna Gorenjak: « Outils biostatistiques /bioinformatiques pour une recherche de médecine personnalisée : télomères, facteur de croissance de l'endothélium vasculaire (VEGF) et molécules associées » Codirected with Dr Sofia Siest Date of ACT: 11/12/2017 Doctoral School BioSE, University of Lorraine Co-financed by the « Région Lorraine », the project « FEDER-FSE Lorraine et Massif des Vosges 2014-2020 » and the project "Agence Nationale de la Recherche, programme d'Investissements d'avenir", grant number ANR-15RHU-0004 Thesis defense: End of 2019 I will participate in the evaluation committee of this thesis. 2. Jérôme Chatelin: « Interactions gène × gène, gène × environnement et Médecine Prédictive, Préventive, Personnalisée et Participative (4P) dans l'inflammation » Codirected with Dr Sofia Siest Date of ACT: 16/09/2015 Doctoral School BioSE, University of Lorraine Thesis defense: February 2019 I will participate in the evaluation committee of this thesis. 3. Ting Xie: « Interactions épistatiques pour le développement des algorithmes de prédiction des maladies cardiovasculaires: De l'approche gène-candidat au séquençage de prochaine génération » Codirected with Dr Sofia Siest Date of ACT: 16/09/2015 Doctoral School BioSE, University of Lorraine Co-financed by the « Région Lorraine », the project « FEDER-FSE Lorraine et Massif des Vosges 2014-2020 » and the project "Agence Nationale de la Recherche, programme d'Investissements d'avenir", grant number ANR-15RHU-0004 Thesis defense: December 2017 I have participated in the evaluation committee of this thesis. Co-direction (50%) of doctoral students without official autorisation 4. Marc Rancier : « Intérêt de la génomique du VEGF en endocrinologie clinique » Codirected with Dr Sofia Siest Doctoral School BioSE, University of Lorraine Thesis defense: March 2015 I have participated in the evaluation committee of this thesis. 5. Abdelsalam Saleh : « VEGF: Un biomarqueur potentiel dans la physiopathologie cardiovasculaire » Codirected with Dr Sofia Siest Doctoral School BioSE, University of Lorraine Financed by Libyan funding Thesis defense: March 2015 I have participated in the evaluation committee of this thesis.

  I have presented these results as oral presentations (conferences 23 and 31) which received the award for Best Research Study in 2007 and 2010 from the Hellenic Society for Research of Metabolic Bone Diseases (awards 5 and 2).

  the polymorphisms of VDR gene, we have identified significant associations between Taq-I and Bsm-I with the presence of osteoporosis [odds ratio (OR)=2.141, 95% confidence interval (95% CI)=1.098-4.174, P=.025, and OR=2.216, 95% CI=1.141-4.306, P=.019, respectively]

Figure 1 .

 1 Figure 1. Adjusted spine bone mineral density (mean95% confidence interval) by

  and I have received the award for Best Research Study in 2009 from the Hellenic Society for Research of Metabolic Bone Diseases (award 4). My doctoral studies were funded by a scholarship from the State Scholarships Foundation in Greece (award 7) following written examinations that ranked me 2nd in a national competition in 2005 and 2 research findings from the Hellenic Society for Research of Metabolic Bone Diseases (award 6 and 9) on 2007 and 2004. Furthermore, the results of the bibliographic research that I had prepared during my doctoral studies were presented in two reviews focused on nutritional aspects of skeletal health, genetics and gene × environment interactions (reviews 7 and 8). During this period, I participated in the educational programme of the doctoral seminaries of the Laboratory for the Research of the Musculoskeletal System (LRMS) of the Medical School of National and Kapodistrian University of Athens. I had given lectures about the links between nutrition, genetics, their interactions and skeletal health (teaching experience 2 and 3). Furthermore, I have participated in the post-graduate course « Nutrition and development » of the Faculty of Dietetics-Nutrition of the Harokopio University of Athens 2010-2011 with a lecture on « Genetic and skeletal health and development » (teaching experience 1).The recruitment of the post-menopausal population used in my doctoral project resulted in the establishment of the "OSTEOS-Osteoporosis: SNPs To Environment Study". This population participated in the GEFOS/GENOMOS consortium which was a European Union (EU) 7thFramework Package funded project. The originality of this study, in which I was the principal researcher, included the large sample size (among the largest populations of Greek origin) and the assessment of interactions with nutrition, which were limited up to that time. With this population, I have participated in another International Consortium, the ReproGen consortium, where I acted as an analyst and author of one meta-analysis concerning the genetic regulation of menopause age (original publication 30)18 . I was also among the initiators of the OSTEOS II study and population in 2010, which received a funding from the Hellenic Society for Research of Metabolic Bone Diseases (award 3) and the initial results were presented as poster presentations in 2 international conferences (conferences 16 and 17).Moreover, as part of my research training, I participated for 1 year in the recruitment, data collection and analysis of the FP6 Project "ZINCAGE. Nutritional zinc, oxidative stress and immune system: biochemical, genetics and life style factors in healthy aging" which was funded by the EU. Although I was not the principle investigator, my participation offered me the experience of working in a multicentre research program on nutritional and genetic epidemiology.In conclusion, this thesis brought together clinician and genetic research scientists and allowed the establishment of strong collaborations in national and international levels. It enforced my skills in laboratory work, genetic epidemiology of human chronic diseases, assessment of genetic interactions with environment, linking of genetics with clinical research as well as efficient group working tasks. It also enforced my skills in genetic epidemiology study designs, statistical analysis, drafting of manuscripts and participating in multicenter projects and international collaborations and consortia. Post-doctoral researcher (2010-2014) Université de Lorraine, Inserm, IGE-PCV « Interactions Gène-Environnement en Physiopathologie Cardio-Vasculaire» U1122 (previously EA 4373 « Génétique Cardiovasculaire »).

Figure 3 :

 3 Figure 3: VEGF, its receptors and their roles. (modified by Vuorio et al, 2012 27 )

Figure 4 .

 4 Figure 4. VEGF gene, isoforms and protein structure 47

  PBMCs and possible genetic links (direct effects and gene × gene interactions) between the 4 VEGF-related SNPs and CAM and inflammation molecules levels in a discovery population of 403 unrelated individuals from SFS. The assessed CAM and inflammation molecules included interleukin 6 (IL6), tumor necrosis factor α (TNF-α), E-selectin, L-selectin, Pselectin, c-reactive protein (CRP) and intercellular adhesion molecule-1 (ICAM-1). Four mRNA isoforms of VEGF gene were quantified (VEGF-121, -145, -165 and -189) and tested for associations with mRNA levels of IL6, TNF-α, L-selectin, P-selectin and ICAM-1. VEGF levels were associated with ICAM-1 (beta = 0.043, P = 0.023) and with E-selectin (beta = 0.085, P = 0.039). The mRNA levels of VEGF-145 isoform were associated with ICAM-1

  a post-doctoral research in the IGE-PCV; U1122 unit, I have participated in numerous research contracts of the unit: 1. "Les Interactions Gène-Environnement dans l'era des analyses pangénomiques (genomewide scan, GWS)". The project was a fellowship for my post-doctoral research funded by the Lorraine Region. My work in the previously detailed projects was included in this contract (2012-2013). 2. "BioIntelligence" project. This was a national French project coordinated by Dassault-Systèmes, with a great number of research and industrial partners: EPI Orpailleur, Sobios, Aureus pharma, Ipsen, Pierre Fabre, Sanofi-Aventis, Servier, Bayer CropScience, INSERM, Genopole Evry. It aimed to the design, development and implementation of clinically useful softwares for life science field and for multifactorial diseases. The research projects of VEGF and gene × gene and gene × environment interactions were used in this project to facilitate the development of metabolic pathways software. Apart from my participation in the research work, I also participated and represented the unit in the frequent meetings of the project group and drafted the annual reports of the team (2010-2014).3. "Gene-Environment Interactions in Cardio-Vascular Physiopathology" project which was funded by the infrastructure BIOBANQUES Inserm under the call « Thematic networks for biobanks ». This project aimed at the development of an informatics platform named "Omics_BioBook" that would be able to manage, store and transfer with safety "-omics" data coming from the BRC IGE-PCV through the collaborating network of the unit IGE-PCV; U1122. I was among the principal researchers participating in this project and I have contributed in the development of this platform with the collaboration of the infrastructure BIOBANQUES and the Inserm Grand Est (2013-2015). 4. ""Omics" data of SFS" project. This project was based on contract between the unit IGE-PCV; U1122 and the Wellcome Trust Sanger Institute in UK that included the generation of GWAS data, epigenome-wide data (whole genome DNA methylation assay) and RNA sequencing data for a selected familial sample of SFS. I was involved in this project in the selection of the individuals, in transfer of biological material and data and in data analysis and interpretation (2013-2014). The results of this collaboration will be developed in the next section (Research Engineer Inserm). 5. "TELARTA (Telomeres and Arterial Aging)" project. As mentioned before, this was an "ANR Programme de Recherche Translationnelle en Santé", where the unit IGE-PCV; U1122 was responsible for a work package. My participation was detailed above in the project "Telomere length genetic determinants in adults and children" (2014-2017).Supervision of students and participation in evaluation committeesI have co-supervised the thesis of Mohsen Azimi Nezhad: « VEGF-A et phénotypes intermédiaires des maladies cardiovasculaires : une approche de génomique fonctionnelle » (doctoral student 6), whose results were detailed above in the Project 1 : Study of VEGF and its involvement in CVD. The thesis defense was on Septembre 2012.

1 .

 1 Debette S*, Visvikis-Siest S*, Chen MH, Ndiaye NC, Song C, DeStefano AL, Safa R, Azimi-Nezhad M, Sawyer D, Marteau JB, Xanthakis V, Siest G, Sullivan L, Pfister M, Smith H, Choi SH, Lamont J, Lind L, Yang Q, Fitzgerald P, Ingelsson E, Vasan RS**, Seshadri S**. Identification of cis and trans acting genetic variants explaining up to half the variation in circulating VEGF levels. Circ Res. 2011; 109(5): 554-63. * equal first authors, ** equal last authors. 2. Azimi-Nezhad M, Lambert D, Ottone C, Perrin C, Chapel C, Gaillard G, Pfister M, Masson C, Tabone E, Betsou F, Meyronet D, Ungeheuer MN, Visvikis-Siest S. Influence of pre-analytical variables on VEGF gene expression and circulating protein concentrations. Biopreservation and Biobanking 2012; 10(5):454-61.

  (original publication 2), and my work was focused on study design, data interpretation and reviewing of the manuscript. Studies on the VEGF-related SNPs in French populations have been performed and replications are ongoing, where I am implicated in the study design, the data analysis and interpretation. The first results indicate an association of the rs10738760 with decreased risk for metabolic syndrome (OR=0.47, P=0.010). These results have not been published yet and represent a part of the work of the theses of Mohsen Azimi-Nezhad, Abdelsalam Saleh, Marc Rancier and Jerome Chatelin (doctoral students 2, 4, 5 and 6), which I co-supervised. Furthermore, this subject was the focus of a scientific review, where we have participated in the reviewing of the manuscript 92 (scientific review 4). genes reached EWAS significance (P = 1.08 x 10 -07 to P = 5.64 x 10 -15

1 .

 1 VEGF Consortium: I am the project manager of the consortium and member of the Steering Committee. I am participating in the conception and design of multiple projects (details and analyses of publications in the Project "Study on VEGF" above). Since 2015, I have drafted two H2020 proposals within the frames of the calls H2020-SC1-PM-02-2017: New concepts in patient stratification in 2016 and H2020-SC1-BHC-25-2019: Demonstration pilots for implementation of personalised medicine in healthcare in 2018. 2. CHARGE Consortium: I am the main analyst and contact person for the IGE-PCV; U1122 unit. I have participated in 4 projects:  Meta-GWAS CRP project, where apart from analyses, I was member of the writing committee and I participated in the design of methodology, coordination of communication and data centralization of> 100 cohorts participating (details on publication of Project "Novel genetic determinants of inflammation molecules" above).  Meta-GWAS IL6 project, where I have realised the statistical replication analyses in SFS and other populations of the BRC IGE-PCV, and I have participated in the drafting and reviewing of the manuscript.  Meta-GWAS TNF-α project, where I have realised the statistical replication analyses in SFS and other populations of the BRC IGE-PCV, and I have participated in the drafting and reviewing of the manuscript.

  Risk Group Factors of Chronic Disease Collaborating Group (NCD Risk Factor Collaboration -NCD-RisC): I am the main analyst and contact person for the IGE-PCV; U1122 unit. I have participated in data analysis included in 6 publications (details on publications on Other projects above). Participation in research contracts and contracts as PI Since 2015, I have participated in the following contracts of the IGE-PCV; U1122 unit: 1."Mastic treatment for Healthy obese with NAFLD diagnosis-MAST4HEALTH", which is funded by the European Community within the frames of the project H2020-MSCA-RISE-2015: Marie Sklodowska-Curie Actions Research and Innovation Staff Exchange. This European projects is a multicenter double blind randomised clinical trial, where a Mastiha or a placebo treatment is used in patients with non-alcoholic fatty liver diseases (NAFLD).Mastiha is a natural product of Greece, produced in Chios island, with antioxidant/antiinflammatory and lipid lowering properties. The aim of the intervention is to test the effectiveness of Mastiha supplement as a new non-pharmacologic strategy for NAFLD. The investigation of Mastiha effect will be performed through testing of gene-diet interactions, correlation of genetic and epigenetic markers with metabolomic and intestinal microbiota profiles pre-and post-intervention, while the effectiveness of the intervention will be evaluated via clinical and laboratory markers. The consortium of this European project consists of 13 academic and industrial partners from 7 European countries. I have drafted the participation of the IGE-PCV; U1122 unit in the proposal document and reviewed all related parts. The unit is leader of the work package 4: "Effects of Mastiha on inflammation and adhesion biomarkers and on oxidative stress status indexes". I am responsible for the tasks of our team that includes the development of databases with IL6, IL1A, TNF-α and VEGF gene expressions data pre and post intervention. Within the frames of this project, I have worked as secondee for 9 months in the Chios Mastic Growers Association, in Greece, where I worked on coordinating the randomisation of treatments for the clinical trial and the design and development of the trial and 3 months in BIOTECH VANA, in Spain, where I have worked on data preparation for the development of a software for integration of metadata and "-omics" profiles for indexing, management and interpretation. I have participated in the training of 4 secondees that have worked in the IGE-PCV; U1122 unit as detailed in the co-supervision of students section above (2016-2020).2. "Fight Heart Failure, FIGHT-HF", which is funded by ANR within the frames of the projects "Hospital-University health research" (Recherche Hospitalo-Universitaire en santé, RHU), ANR-15RHU-0004 (2015-2020). This project aims to generate new heart failure classifiers, and to improve patient outcomes with the use of a strategic approach that combines knowledge based on i) underlying mechanisms, ii) "-omics" and imaging bioprofiling, iii) co-morbidities and iv) holistic disease management to define mechanistically relevant and clinically actionable bioprofiles. The consortium is consisted of 8 academic and industrial partners. The IGE-PCV; U1122 unit is responsible for the task 2.3: "TREM-1: inflammation in HF post-myocardial infarction" of work package 2 and more specifically the study of genetic links between TREM-1 with inflammation molecules and for the task 3.1: "-Omics biomarker prediction of transition to heart failure and mechanistic profiling task" of work package 3 and most specifically the study of genetics of TL as a marker of vascular aging. My participation on this contract is detailed in the project "TREM and inflammation" and "Telomere length as a biomarker for personalised medicine" above (2015 -2020).3. "TELARTA" project, which is funded by ANR within the frames of the project "ANR Programme de Recherche Translationnelle en Santé". Details on this contract have been given in the previous section (Post-doctoral research) but novel actions were taken as described in the project "Telomere length as a biomarker for personalised medicine" above (2014 -2017).4. "GEENAGE project (Functional Genomic, Epigenomic and ENvironment interplay toIMPACT management of healthy and pathological AGEing)" funded within the frames of the LUE project (Lorraine University of Excellence). The aim of this contract is to propose new strategies of diagnosis and management of healthy and pathological ageing by targeting the functional consequences of the interplay between genes, epigenome and environment.The IGE-PCV; U112 unit is involved in the scientific research axe 3 : "Regulation of telomere length and impact on the accelerated cardiovascular ageing and relevant comorbidities", which aims to study the genetic and environmental factors that are important in the regulation of telomeres early in life. The work on TL genetics described in the project "Telomere length as a biomarker for personalised medicine" above is my participation in this contract (2016 -2026).5." Development of a system for the management and analysis of biological samples and "omics" data related to VEGF" project which is funded by Pôle BMS « Biologie, Médecine, Santé » within the frames of the project AAP-BMS interne "Projets incitatifs -Crédits CS-UL 2018". For this project I am the Principal Investigator. This scientific project aims, through a network composed by 1 biological resource center (BRC IGE-PCV), 1 Research Unit (IGE-PCV; U1122) and 1 International Consortium (VEGF Consortium), the development of a system for management and analysis of biological samples and associated "-omics" data to elucidate factors that are linked with VEGF and which are involved in the development of cardiovascular pathologies. The project is linked with "Omics_BioBook" platform, which is a biostatistical/bioinformatics platform where associated data are stored and managed. Management software for samples and data are under development in order to facilitate and amplify the interconnections and exchanges between network partners (2018-2019).

2 .

 2 Jérôme Chatelin: « Interactions gène x gène, gène x environnement et Médecine Prédictive, Préventive, Personnalisée et Participative (4P) dans l'inflammation ». The defense will take place at the beginning of 2019 and I will participate in the evaluation committee (doctoral student 2).

1 .

 1 Stathopoulou MG, Xie T, Ruggiero D, Chatelin J, Rancier M, Weryha G, Kurth MJ, Aldasoro Arguinano AA, Gorenjak V, Petrelis AM, Dagher G, Dedoussis G, Deloukas P, Lamont J, Marc J, Simmaco M, Van Schaik RHN, Innocenti F, Merlin J-L, Schneider J, Alizadeh BZ, Ciullo M, Seshadri S, Visvikis-Siest S -The VEGF Consortium. A transnational collaborative network dedicated to the study and applications of the Vascular Endothelial Growth Factor-A in medical practice: The VEGF Consortium. Clin Chem Lab Med. 2018 Mar 28;56(4):83-86. 2. Chatelin J*, Stathopoulou MG*, Aldasoro Arguinano AA*, Xie T*, Visvikis-Siest S*. Pharmacogenomic challenges in cardiovascular diseases: examples of drugs and considerations for future integration in clinical practice. Curr Pharm Biotechnol. 2017;18(3):231-241. * equal first authors 3. Rancier M*, Zaaber I*, Stathopoulou MG, Chatelin J, Saleh A, Marmouch H, El Shamieh S, Masson C, Murray H, Lamont J, Fitzgerald P, Mahjoub S, Said K, Tensaout BB, Mestiri S, Visvikis-Siest S. Pro-and anti-angiogenic VEGF mRNAs in autoimmune thyroid diseases. Autoimmunity. 2016 Aug 5:1-7. * equal first authors 3. Vesna Gorenjak: « Outils biostatistiques /bioinformatiques pour une recherche de médecine personnalisée : télomères, facteur de croissance de l'endothélium vasculaire (VEGF) et molécules associées ». The defense is planned for the end of 2019 I will participate in the evaluation committee (doctoral student 1).

4 .Other activities linked to research 1 .

 41 Gorenjak V, Akbar S, Stathopoulou MG, Visvikis-Siest S. The future of telomere length in personalized medicine. Front Biosci (Landmark Ed). 2018 Mar 1;23:1628-1654. Furthermore, I was the professional tutor of the trainee of License Professionel of Chloé Hatrisse : « Certification d'un Centre de Ressources Biologiques » (trainee 2). The aim of this internship was the support of the procedures for the development and the quality certification of the Quality Management System of the BRC IGE-PCV. Also, during the project « Mastiha treatment for obese with NAFLD diagnosis -MAST4HEALTH », within the framework of the HORIZON 2020 Marie Skłodowska-Curie actions -Research and Innovation Staff Exchange (MSCA RISE), I was the responsible for the training of 4 secondees in the fields of genetic epidemiology and related statistical analyses: Dwaine Vance, Randox Laboratories, 6 months, Lauren Doherty, Randox Laboratories, 3months, Eirini Chroni, Chios Mastiha Growers Association, 5 months and Georgia Maurikou, Chios Mastiha Growers Association, 5 months (trainnees 1). Organisation of conferences I have participated in the organisation of the following conferences as member of the organising committee:  The 9th Santorini Conference, Santorini, Greece, 2018.  8th Santorini Conference, Santorini, Greece, 2016.  Third ESPT Conference: Integration of pharmacogenomics in clinical decision support, Budapest, Hungary, 2015. My tasks included the communication of the conferences, the organisation of poster sessions, on site organisation and the drafting of the post-conference scientific manuscripts. In the 8th Santorini Conference I was also member of the Poster Committee that was responsible for the evaluation of posters and selection of award winners. I was President of this committee in the 9th Santorini Conference. BRC IGE-PCV I am member of the Steering Committee of the BRC IGE-PCV (BB-0033-00051) located in the IGE-PCV; U1122 unit. I am Manager and Coordinator of Collaborations and Responsible for the transfert of data and biological material. I ensure all the processes concerning the inputs and outputs, the transfer and the quality of the samples and stored data. I coordinate the release and the entry of data and the development and management of the related databases. I am also Quality advisor and I participate in the implementation of the quality approach for the quality certification according to the standard NF S 96-900. I participate in the development of the Quality Management System, in the meetings and the audits related to the quality certification.

  collaboration with the Inserm Grand Est, the University of Lorraine and the infrastructure BIOBANQUES. The platform is installed in the Interuniversity Center of Informatics Resources of Lorraine -C.I.R.I.L (Centre interuniversitaire de ressources informatiques de Lorraine). It is a platform of high-performance with high storage capacity (60 TB for storage and 80 To in the automatic replication server) that is designed so as to meet the methodological and technological advances of "-omics" data storage and analysis, to ensure the security of the data of the BRC IGE-PCV, and to ensure the speed and performance of the large-scale analyses of the international consortia in which the IGE-PCV; U1122 unit participates. I have participated very actively in its development and I am the platform administrator concerning the management of user sessions. I participate in the development of a customized version of the software TumoroteK® which is used for the management of biological samples in the "Omics_BioBook" platform. An interface is also being developed

Figure 6 :

 6 Figure 6: Role of VEGFs in the normal vessel wall and atherosclerotic lesions.

  and osteoblasts, nutrients (calcium, phosphorus, protein), vitamin D, growth and differentiation factors200, 201 . VEGF receptors on ECs are involved with the activation of expression of cell-adhesion molecules, release of cytokines and chemokines resulting in degradation of the vessel's basement membrane, allowing cells to migrate to the extracellular matrix202 . VEGF receptors are present on primary osteoblasts 203 and VEGF promotes bone formation, acting directly on survival, chemotactic migration 204 and the activity 205 of osteoblasts. Mice that do not express VEGF in bone progenitor cells present an osteoporosis phenotype with decreased trabecular bone, thickening of bone cortices and increased adipocytes associated with trabecular bone[START_REF] Mangino | Genome-wide meta-analysis points to ctc1 and znf676 as genes regulating telomere homeostasis in humans[END_REF] . All of these observations provide evidence of the major role VEGF-A plays in bone development and remodeling.The IGE-PCV; U1122 unit's main objective is to determine the respective roles of genetic and environmental factors (and their interactions) involved in the development of and predisposition to CVD. In order to identify early, specific biomarkers, the unit has set up a scientific strategy based on an intermediate phenotype-genotype approach and used family, cross-sectional and longitudinal study designs. This methodological strategy is unique in integrating genomics, transcriptomics and proteomics (systems biology), thus facilitating the identification of functional polymorphisms with significant use as predictive and/or preventive markers. VEGF is one of the major axes of the research work of the unit. The majority of the work completed so far on VEGF by the unit, including the studies developed in the section above where I have participated, are summarized in the figure7.

Figure 7 .

 7 Figure 7. Realisations of the IGE-PCV; U1122 unit with my contribution on VEGF.

  Member of the emergency standby personnel for the security of biological material stored in the freezers of the BRC IGE-PCV.

	Period : 01/04/2014 -30/09/2014
	I have participated in the evaluation committee of this thesis.
	Trainees (stagiaires)
	1. Responsible for training of 4 secondees participating in the project « Mastiha treatment for Awards and Fellowships
	obese with NAFLD diagnosis -MAST4HEALTH », within the framework of the HORIZON
	2020 Marie Skłodowska-Curie actions -Research and Innovation Staff Exchange (MSCA 1. 2012 Fellowship for post-doctoral research entitled: "Les Interactions Gène-
	RISE):	Environnement dans l'era des analyses pangénomiques (genome-wide scan, GWS)",
		 Dr Dwaine Vance, Randox Laboratories, period: 02/10/2017 -02/04/2018 Région Lorraine, France.
	2.	 Lauren Doherty, Randox Laboratories, period: 02/10/2017 -02/01/2018 2010 Award for Best Research Study: "Association of VDR polymorphisms with
		 Eirini Chroni, Chios Mastiha Growers Association, period: 01/04/2018 -31/08/2018 peak bone mass in Greek men", Hellenic Society for Research of Metabolic Bone
		 Georgia Maurikou, Chios Mastiha Growers Association, period: 01/04/2018 -Diseases, Greece.
	3.		31/08/2018
	Codirected with Dr Sofia Siest
	Doctoral School BioSE, University of Lorraine
	Financed by Iranian funding
	Thesis defense: September 2012
	7.		2005	Fellowship for doctoral studies from the State Scholarships Foundation,
	Co-direction (50%) of master students Greece
	8.		2004	Award for Best Research Study: "VDR polymorphism is related to an
	1. Alexandros Petrelis : « Investigations biostatistiques/bioinformatiques des interactions gène increased risk of vertebral fractures in postmenopausal osteoporotic women",
	× gène impliquées dans le risque cardiovasculaire -la longueur des télomères leucocytaires Hellenic Society for Research of Metabolic Bone Diseases, Greece.
	chez les enfants » 9. 2004 Funding for the research program entitled: "Interaction of nutritional and
	Codirected with Dr Sofia Siest
	Master II Ingénierie Mathématique et Outils Informatiques, IMOI, Faculty of Sciences and
	Technologies, University of Lorraine

6. Mohsen Azimi Nezhad : « VEGF-A et phénotypes intermédiaires des maladies cardiovasculaires : une approche de génomique fonctionnelle » 2. Professional tutor (50% with Christine Masson) of Chloé Hatrisse : « Certification d'un Centre de Ressources Biologiques » License Professionnel « Biotechnologies appliquées à la santé », University of Reims Champagne-Ardenne) Period : 30 January 2017 -28 July 2017 Other activities linked to research 1. Member of the Steering Committee of the Biological Resources Centre (BRC) IGE-PCV (BB-0033-00051) of the Unit IGE-PCV; U1122, Manager and Coordinator of Collaborations, Responsible for the transfert of data and biological material, and Quality advisor 2. Administrator of the "Omics_BioBook" Biostatistics / Bioinfomatics platform Administrative activities Member of the laboratory council of the unit IGE-PCV; U1122 Other activities 2010 Funding for the research program entitled: "Vitamin D levels, bone markers and genetic factors in Greek sample of men and women", Hellenic Society for Research of Metabolic Bone Diseases, Greece. 4. 2009 Award for Best Research Study "The effect of common polymorphisms of LRP5 gene on bone mineral density of Greek postmenopausal women", Hellenic Society for Research of Metabolic Bone Diseases, Greece 5. 2007 Award for Best Research Study: "Interaction of nutrition and environmental factors with genetic polymorphisms of the VDR and CTR gene on peak bone mass in Greek men", Hellenic Society for Research of Metabolic Bone Diseases, Greece. 6. 2007 Funding for the research program entitled: "Association study of genetic polymorphisms with bone mineral density and risk factors for osteoporosis", Hellenic Society for Research of Metabolic Bone Diseases, Greece. environmental factors with genetic polymorphisms of the VDR and CTR gene on peak bone mass in Greek men", Hellenic Society for Research of Metabolic Bone Diseases, Greece.

Résumé des travaux de recherche, d'enseignement et d'administration en français

  Pendant mes études (Maîtrise, bac+4) en nutrition clinique et en diététique à l'Université Harokopio d'Athènes, en Grèce, j'ai été attiré par le domaine de la génétique et des interactions avec la nutrition. Mon projet de thèse a été réalisé au laboratoire de recherche sur le système musculo-squelettique (LRMS) de la Faculté de Médecine de l'Université Nationale et Kapodistrienne d'Athènes (Grèce) et s'intitulait «Réponse au traitement par la calcitonine et association au polymorphisme de la CTR chez la femme ménopausée ostéoporotique». Après mes études et en parallèle à mon doctorat, j'ai été recrutée comme chercheur contractuel à l'Université Harokopio d'Athènes, en Grèce, et j'ai travaillé dans 9 projets de recherche concernant les effets de la nutrition sur la santé et les maladies chroniques. Ces projets comprenaient 4 essais cliniques randomisés, 1 étude d'intervention et 4 études observationnelles. Ils ont abouti à 8 articles originaux (publications 36, 37, 38, 39, 40, 41, 43, 44) et ils ont été présentés dans 3 conférences (conférences 22, 29, 30). . L'identification des déterminants «-omiques» du VEGF qui affectent le risque de MCV, de AD et d'ostéoporose dans les populations cas-témoins et les études prospectives. Cette approche permettra de tester la spécificité de différents déterminants «-omiques» sur

	appliquée dans GWAS) peuvent également être ajoutés à la liste des gènes candidats risque calculé par le système SCORE sur dix ans d'un premier événement MCV fatal, telle
	déterminants du VEGF. De nouvelles découvertes seront également prévues par les études qu'une crise cardiaque, un accident vasculaire cérébral, un anévrisme de l'aorte ou un autre
	travail d'interaction gène × environnement. Des réplications seront réalisées au UK Biobank. Un événement sera également utilisé comme phénotype d'intérêt. Ces phénotypes sont
	différentes maladies. score génétique de tous les marqueurs indépendants sera développé. disponibles dans les cohortes du Consortium VEGF, CHARGE et la UK Biobank, réunissant
	J'ai co-dirigé la thèse de Mohsen Azimi Nezhad: «VEGF-A et phénotypes intermédiaires des molécules associées». La soutenance est prévue pour fin 2019 et je participerais au comité 4. Le développement de modèles et d'algorithmes de prédiction du risque basés sur les VEGF un échantillon puissant. Les interactions directes, gène × gène et gène × environnement seront
	Pendant la période de post-doctorant à l'UMR INSERM; IGE-PCV «Interactions Gène maladies cardiovasculaires: une approche de la génomique fonctionnelle». La soutenance de d'évaluation. Jusqu'à présent, 4 publications ont été produites par cette thèse. «-omiques». Leur valeur prédictive sera évaluée dans le cadre d'études prospectives visant à Déterminants transcriptomiques des taux de VEGF testées. Cet axe identifiera des déterminants spécifiques «-omiques» du VEGF qui affectent
	Environnement-Physio-Cardio-Vasculaire» j'ai travaillé dans 5 thématiques de recherche: la thèse a eu lieu en septembre 2012 et a donné lieu à 6 publications. Je suis membre du conseil de laboratoire de l'Unité IGE-PCV; U1122. Je participe aux identifier les groupes à risque parmi les individus tout au long de leur vie. Leur utilité et Le séquençage de l'ARN par TrueSeq (Illumina) sur des instruments HiSeq2500 (75 pb paired des voies spécifiques et des facteurs de risque de maladies.
	5) Second ESPT Conference Pharmacogenomics: From Cell to Clinic, Lisbon, Portugal, • Projet 1: Etude du facteur de croissance de l'endothélium vasculaire (VEGF) et de son J'ai co-dirigé la thèse de Marc Rancier : «Intérêt de la génomique du VEGF en endocrinologie J'étais le tuteur professionnel du stagiaire de la Licence Professionnel Chloé Hatrisse: réunions annuelles qui ont pour objectif de planifier les décisions stratégiques pour l'avenir de l'efficacité de la stratification seront comparées aux algorithmes conventionnels de prévision end reads) a été effectué pour 220 personnes appartenant à 70 familles de la cohorte SFS. Ces
	2013. implication dans les maladies cardiovasculaires (MCV) clinique» et Abdelsalam Saleh: «VEGF: Un biomarqueur potentiel dans la physiopathologie «Certification d'un Centre de Ressources Biologiques». Dans le cadre du projet H2020 MSCA l'unité. du risque, en tenant compte à la fois de l'âge et du sexe. données et d'autres données transcriptomiques par la méthode de séquençage d'ARN ou de Axe 3: Identification des déterminants moléculaires du VEGF associés au risque de maladies
	6) 6th Santorini Conference Biologie Prospective, Santorini, Greece, 2012. • Projet 2: Évaluation des interactions gène × gène et gène × environnement pour la cardiovasculaire» qui ont été présentées en mars 2015 et qui ont donné lieu respectivement à 4 RISE 2015, j'étais responsable pour la formation de 4 personnes dans le cadre du projet Le projet utilisera les ressources disponibles dans l'IGE-PCV; U1122 et le consortium VEGF qPCR disponibles dans les populations saines participant au consortium VEGF seront chroniques
	régulation de la pression artérielle et 3 publications. J'ai participé au comité d'évaluation de ces deux thèses. «MAST4HEALTH» pour les domaines de l'épidémiologie génétique et des analyses coordonné par l'Unité. Les ressources en population incluent des cohortes reconnues (SFS, combinées afin de quantifier l'expression des isoformes pro-et anti-angiogéniques du gène du L'objectif principal de l'axe 3 est d'identifier les déterminants « -omiques » du VEGF qui
	VEGF dans les cellules sanguines mononucléaires périphériques. Après leur quantification, pourraient être utilisés pour prédire le risque de maladie. On utilisera des populations cas-
	Member of Scientific Societies une analyse d'association et des méta-analyses seront effectués afin d'identifier les isoformes témoins atteintes de MCV, d'AD et d'ostéoporose. Ces populations sont disponibles dans le
	Hellenic Osteoporosis Foundation qui affectent davantage les niveaux de VEGF. CRB IGE-PCV, les consortia VEGF, CHARGE, METASTROKE et MEGASTROKE, et les
	European Society of Pharmagenomics and Personalised Therapy biobanques BBMRI-ERIC et UK Biobank. Dans ces populations, les déterminants «-
	European Society of Predictive Medicine Déterminants épigénétiques des taux de VEGF omiques» du VEGF identifiés dans les axes 1 et 2 seront testés pour déterminer les
	Le profil de méthylation de l'ADN à l'aide de la puce Infinium Methylation450K (Illumina) associations avec un risque accru de maladies évaluées. Nous nous attendons à identifier des
	capable d'interroger 450000 sites CpG et de quantifier les niveaux de méthylation de l'ADN déterminants «-omiques» spécifiques pour chaque maladie et à conduire à la sélection des
	est disponible pour 220 personnes de la cohorte SFS et des autres populations en bonne santé biomarqueurs les plus puissants à ajouter aux modèles de prévision du risque du prochain axe.
	participant au consortium VEGF. Des méta-analyses seront effectuées pour identifier les
	sondes de méthylation associées de manière significative aux niveaux de VEGF. Axe 4: Développement et évaluation de la valeur prédictive d'algorithmes personnalisés de
	prédiction du risque basés sur les déterminants «-omiques» du VEGF
	Axe 2: Identification des déterminants « -omiques » du VEGF associés aux facteurs de risque L'objectif principal de l'axe 4 est de développer et de démontrer la valeur prédictive
	U1122. Jusqu'à aujourd'hui, je participe à 5 projets: • Projet 1: L'étude sur le VEGF • Projet 2: Effets des polymorphismes du LSR sur la lipidémie et la maladie d'Alzheimer • Projet 3: Récepteurs déclencheurs exprimés sur les cellules myéloïdes (TREM) et l'inflammation Déterminants génétiques des taux de VEGF Nouvelles méta-analyses pangénomiques (GWAS) seront utilisées avec l'ajout de nouvelles populations disponibles via le consortium CHARGE et l'étude ORIGIN. L'objectif est de rassembler plus de 20000 personnes, augmentant ainsi la puissance de l'étude et le potentiel d'identification de nouveaux variants qui augmenteront le pourcentage de variabilité des correction FDR (critère moins strict comparé à la correction classique de Bonferroni indice de produit d'accumulation de lipides (LAP) et indice d'adiposité viscérale (VAI). Le développée, tandis que des polymorphismes atteignant un niveau de signification de 0,05 par taille / hanche, les lipoprotéines de haute et bas densité, l'apolipoprotéine E, les triglycérides, niveaux de VEGF expliqués. Une liste de polymorphismes indépendants significatifs sera indépendante et additive de déterminants du VEGF spécifiques aux MCV, à l'AD et à de maladies chroniques l'ostéoporose, identifiés dans les axes précédents pour l'évaluation du risque de maladie par L'objectif principal de l'axe 2 est d'établir et de quantifier l'effet des déterminants «-omiques» des algorithmes de prédiction computationnels. La valeur prédictive sera évaluée dans de du VEGF sur les facteurs de risque des maladies chroniques et leurs interactions avec les grandes cohortes prospectives dans le but d'identifier les groupes à risque parmi les individus facteurs environnementaux. Ces facteurs de risque comprennent les lipides sanguins, la tout au long de leur vie. L'utilité prédictive incrémentielle et l'efficacité de stratification des pression artérielle, le métabolisme du glucose, l'obésité et les traits d'inflammation : la nouveaux algorithmes seront comparées aux algorithmes classiques de prédiction du risque pression artérielle systolique, la pression artérielle diastolique, la pression artérielle moyenne, tels que le système SCORE, le risque cardiovasculaire de Framingham (FGCVDRP) pour les la protéine C réactive, l'haptoglobine, l'interleukine 6, le facteur de nécrose tumorale α, la glycémie à jeun, les taux d'insuline, l'indice de masse corporelle, la tour de taille, le rapport MCV et l'outil d'évaluation du risque de fracture FRAX WHO pour l'ostéoporose.

Mon introduction dans le domaine de la recherche en épidémiologie génétique des maladies chroniques a été efficace pendant mon doctorat grâce à mes projets de recherche centrés sur l'ostéoporose au Laboratoire LRMS de la Faculté de Médecine de l'Université Nationale et Kapodistrienne d'Athènes en Grèce. Les phénotypes liés à l'ostéoporose sont hautement héritables (par exemple, l'héritabilité de la densité minérale osseuse (DMO) est estimée à 50-80%) et, comme la plupart des maladies chroniques, elle est caractérisée par une régulation polygénique. Les études pangénomiques (GWAS) et gènes-candidats ont réussi à identifier des variants génétiques associés à la DMO et de nouvelles voies biologiques menant à de nouveaux agents thérapeutiques. L'objectif de ma recherche de master II et de doctorat était d'évaluer les variants génétiques associées à la DMO, en tenant compte les interactions possibles avec apport alimentaire en calcium. Trois gènes-candidats ont été sélectionnés à cause de leur implication dans des voies pathophysiologiques classiques et nouvelles d'ostéoporose, le récepteur de la vitamine D (VDR), le récepteur de la calcitonine (CTR) et la protéine 5 liée aux récepteurs de lipoprotéines de basse densité (LRP5). J'ai démontré que les polymorphismes de gènes CTR, VDR et LRP5 ne déterminent pas la masse osseuse chez les hommes jeunes (publication originale 42) et qu'ils ont un effet important sur la DMO uniquement lorsque la consommation de calcium est faible chez les femmes ménopausées. Par contre la consommation élevée de calcium a un effet protecteur sur la DMO contre la détermination génétique. Nous avons également souligné l'importance de prendre en compte l'apport nutritionnel et les interactions de la nutrition avec les polymorphismes génétiques lors de l'évaluation du risque de maladies multifactorielles chroniques telles que l'ostéoporose. Les résultats correspondants ont été publiés dans deux publications originales (publications originales 34 et 35). • Projet 3: Etude d'association pangénomique sur les déterminants génétiques des taux d'haptoglobine • Projet 4: Déterminants génétiques de la longueur des télomères chez l'adulte et l'enfant • Projet 5: Projets pharmagénomiques du clopidogrel J'ai participé à de nombreux contrats de recherche de l'Unité: 1. «Les interactions génétiques dans l'analyse des études pangénomiques» en bénéficiant d'une bourse par la Région Lorraine pour ma recherche post-doctorale 2. Projet «BioIntelligence». Il s'agissait d'un projet national coordonné par Dassault-Systèmes et regroupant de nombreux partenaires de recherche et industriels: EPI Orpailleur, Sobios, Aureus Pharma, Ipsen, Pierre Fabre, Sanofi-Aventis, Servier, Bayer CropScience, INSERM, Génopole Evry. Il visait à concevoir, développer et mettre en oeuvre des logiciels utiles sur le plan clinique pour les sciences de la vie et les maladies multifactorielles. Les projets de recherche sur le VEGF et les interactions gène × gène et gène × environnement ont été utilisés dans le cadre de ce projet pour faciliter le développement de logiciels de voies métaboliques 3. Projet «Interactions gènes-environnement en physiopathologie cardio-vasculaire» financé par l'infrastructure BIOBANQUES Inserm dans le cadre de l'appel «Réseaux thématiques de biobanques». Ce projet visait à développer une plate-forme informatique appelée «Omics_BioBook», capable de gérer, stocker et transférer avec la «sécurité» les données provenant du BRC IGE-PCV via le réseau collaborateur de l'unité IGE-PCV; U1122 4. Projet «Omics» de STANISLAS Family Study. Ce projet reposait sur un contrat entre l'unité IGE-PCV; U1122 et le Wellcome Trust Sanger Institute au Royaume-Uni 5. Projet «TELARTA (Télomères et vieillissement artériel)». Il s'agissait d'un programme de recherche financé par l'ANR, où l'unité IGE-PCV; U1122 est responsable d'un package de J'ai co-supervisé le master II d'Alexandros Petrelis: «Investigations biostatistiques / bioinformatiques des interactions entre gènes impliquées dans le risque cardiovasculairelongueur des télomères leucocytaires chez les enfants». La défense était en septembre 2014 et a abouti à une publication originale. J'ai participé à l'organisation des conférences suivantes en tant que membres du comité d'organisation: • 7ème Conférence Biologie Prospective de Santorin, Santorin, Grèce, 2014. • 2ème Conférence ESPT Pharmacogénomique: De la cellule à la clinique, Lisbonne, Portugal, 2013. • 6ème Conférence Biologie Prospective de Santorin, Santorin, Grèce, 2012. Depuis 2015, j'ai été recrutée comme ingénieur de recherche à l'INSERM au sein de l'UMR • Projet 4: La longueur des télomères en tant que biomarqueur pour la médecine personnalisée • Projet 5: Nouveaux déterminants génétiques des molécules d'inflammation publications à ce jour. La soutenance aura lieu en 2019 et je participerais au comité d'évaluation. 3. Vesna Gorenjak: «Outils biostatistiques /bioinformatiques pour une recherche de médecine personnalisée : télomères, facteur de croissance de l'endothélium vasculaire (VEGF) et statistiques: Dwaine Vance, Laboratoires Randox, 6 mois, Lauren Doherty, Laboratoires Randox, 3 mois, Eirini Chroni, Association des producteurs de Chios Mastiha, 5 mois et Georgia Maurikou, Association des producteurs de Chios Mastiha, 5 mois. J'ai participé à l'organisation des conférences suivantes en tant que membre du comité d'organisation: • 9ème Conférence de Santorin, Santorin, Grèce, 2018. • 8ème Conférence de Santorin, Santorin, Grèce, 2016. • Troisième conférence ESPT: Intégration de la pharmacogénomique dans l'aide à la décision clinique, Budapest, Hongrie, 2015. Lors de la 8ème Conférence de Santorin, j'étais également membre du comité des posters, chargé de l'évaluation des posters et de la sélection des lauréats pour les prix. J'étais président de ce comité lors de la 9ème Conférence de Santorin. Je suis également membre du comité de direction du Centre de Ressources Biologiques (CRB) IGE-PCV (BB-0033-00051) situé dans l'Unité IGE-PCV; U1122. Je suis responsable et coordinatrice des collaborations et responsable du transfert des données et du matériel biologique. Je suis également référent qualité et je participe à la mise en oeuvre de la démarche qualité de la certification selon la norme NF S 96-900. Je suis également l'administrateur de la plate-forme «Omics_BioBook». Cette plateforme est une plateforme biostatistique / bioinformatique, développée en collaboration avec l'Inserm Grand Est, l'Université de Lorraine et l'infrastructure BIOBANQUES. Il s'agit d'une plateforme de haute performance dotée d'une capacité de stockage élevée (60 To pour le stockage et 80 To pour le serveur de réplication automatique), conçue pour répondre aux avancées méthodologiques et technologiques du stockage et de l'analyse de données «-omiques», afin de garantir la sécurité des données du CRB IGE-PCV, et d'assurer la rapidité et la performance des analyses à grande échelle des consortiums internationaux dans lesquels l'IGE-PCV; U1122 participe. Résume du projet de recherche en français L'étude des déterminants « -omiques » du VEGF dans la médecine personnalisée des maladies chroniques Objectifs L'objectif général du projet est de développer des algorithmes de prédiction du risque pour la prédiction personnalisée du risque des MCV et d'autres maladies chroniques sur la base des déterminants «-omiques» du VEGF. Des données volumineuses provenant de déterminants «omiques» (génétique, transcriptomique, protéomique et épigénétique) du VEGF et des molécules associées seront combinées avec les facteurs environnementaux et avec des interactions épistatiques avec d'autres molécules impliquées dans les processus physiopathologiques communs aux maladies chroniques (angiogenèse, inflammation, métabolisme des lipides). Les objectifs spécifiques du projet sont: 1. L'identification des déterminants «-omiques» du VEGF et de leurs interactions avec les facteurs environnementaux en santé par le biais de méthodes multidisciplinaires «-omiques» robustes et valables (y compris la génétique, l'épigénétique et la transcriptomique) dans de grandes populations longitudinales et familiales saines d'ascendance européenne. 2. Identification des déterminants «-omiques» du VEGF qui affectent les facteurs de risque de MCV, de la maladie d'Alzheimer (AD) et de l'ostéoporose, ainsi que leurs interactions avec les facteurs environnementaux chez des populations saines.

3Framingham Heart Study (FHS); étude Cilento, LLs, ORIGIN), des populations cas-témoins, des populations issues des biobanques (BRC IGE-PCV, BBMRI-ERIC, une infrastructure paneuropéenne, et UK Biobank) et des collaborations avec d'autres consortiums tels que CHARGE, METASTROKE et MEGASTROKE pour lesquels les données «-omiques» souhaitées sont disponibles. La plate-forme «Omics_BioBook» sera utilisée pour le stockage et l'analyse des données. Mise en oeuvre du projet Le projet sera basé sur 4 axes, un pour chaque objectif spécifique. Axe 1: Identification des déterminants «-omiques» du VEGF en santé L'objectif principal de l'axe 1 est d'identifier les déterminants génétiques, transcriptomiques et épigénétiques de la biologie du VEGF en santé en utilisant de méthodes «-omiques» robustes et validés dans de grandes cohortes familiales et longitudinales d'individus en bonne santé.

Research, teaching and administrative activities Assistant Researcher (2002-2005) in the project: Response in calcitonin therapy and association with CTR polymorphisms in osteoporotic postmenopausal women. Medical School of Athens, National and Kapodistrian University of Athens, Harokopio University of Athens, Greece

	During my bachelor studies in clinical nutrition and dietetics (4 years university studies) in
	Harokopio University, Athens, Greece, I first came in contact and was attracted by the field of
	genetics and interactions with nutrition. My bachelor dissertation project was performed in the
	Laboratory for the Research of the Musculoskeletal System (LRMS), Medical School,
	National and Kapodistrian University of Athens, Greece and was entitled "Response in
	calcitonin therapy and association with CTR polymorphisms in osteoporotic postmenopausal
	women".
	The study was performed in osteoporotic postmenopausal women treated with nasal calcitonin
	and alphacalcidol combined therapy (n=160). Selected candidate genes implicated in classical
	and novel osteoporosis pathways were tested including the vitamin D receptor (VDR)
	Nombre de publications originales (revues avec comité de lecture): 44 (rs1544410) and the calcitonin receptor (CTR) (rs1801197).  Nombre de revues scientifiques (revues à comité de lecture): 9 The study indicated the VDR polymorphism was associated with an increased risk of vertebral  Nombre de publications des conférences (revues à comité de lecture): 13 fracture. Furthermore, none of the polymorphisms affected the response to calcitonin therapy.
	Conférences-colloques: 34 These results were presented in 3 conferences (conferences 32, 33 and 34) and received one
	award for Best Research Study in 2004 from the Hellenic Society for Research of Metabolic
	Sur les 53 publications originales et revues scientifiques, 48 ont été trouvées dans la base de Bone Diseases (award 8).
	données Web Of Science en novembre 2018. Les analyses de ces 48 publications ont révélé This study was funded by Pharmaserve Lilly S.A.C.I.
	ce qui suit: My participation included the DNA isolation and genotyping and databases preparation. This
	Somme des citations: 3528 work introduced me to genetic epidemiology, clinical research and human chronic diseases
	Somme des citations excluant les auto-citations: 3482 fields.
	Moyenne de citations par article: 73.5
	Indice h: 16

Contractual researcher (2004-2010) Harokopio University of Athens, Greece

	Following my bachelor studies and in parallel with my doctorate, I was recruited as a
	contractual researcher in Harokopio University of Athens, Greece, in 9 research projects
	concerning the effect of nutrition in health and disease. These projects included 4 randomized
	clinical trials, 1 intervention study and 4 observational studies. They have resulted in 8
	original papers (publications 36, 37, 38, 39, 40, 41, 43, 44) 1-8 and they have been presented in
	3 conferences (conferences 22, 29, 30).
	My contribution in these studies included patients' recruitment, data collection, analysis and
	interpretation of data and drafting of the manuscripts. As these projects are not directly linked
	with my research interests on genetic epidemiology are not presented in details in this report.
	However, their input in my research experience was significant as they enhanced my
	experience in multidisciplinary fields, including different study design, statistical analysis and
	data interpretation and they increased my skills in clinical research.

Doctoral researcher (2004-2010): Assessment of genetic variants associated with bone mineral density in Greek populations and gene × environment interactions Laboratory for the Research of the Musculoskeletal System (LRMS), Medical School, National and Kapodistrian University of Athens, Greece

	My introduction in the field of genetics epidemiology research was effective through my
	master and doctoral research projects focused on osteoporosis. Osteoporosis is a chronic
	disease characterized by deterioration of bone mass and increased susceptibility to fragility
	fractures 9 . It causes an extreme health and economic burden. It is responsible for more than
	8.9 million fractures annually and an osteoporotic fracture happens every 3 seconds.
	Osteoporotic fractures are linked with decrease mobility, functionality and quality of life and
	with increased mortality and morbidity. Also, given that by 2050, the worldwide incidence of
	hip fracture is projected to increase by 310% in men and 240% in women, it is clear that it
	represents a major public health issue problem 10 (http://www.iofbonehealth.org/facts-
	statistics). Osteoporosis related traits are highly heritable (e.g. bone mineral density (BMD)

Research Engineer Inserm, IR2 (2015 -today) Université de Lorraine, Inserm, IGE-PCV; U1122 Project 1 : The study on VEGF

  The defense was on September 2014 and I have participated in the evaluation committee.

	5. Azimi-Nezhad M*, Stathopoulou MG*, Bonnefond A, Rancier M, Saleh A, Lamont J, Mestiri S, Visvikis-Siest S. Pro-and anti-angiogenic VEGF mRNAs in autoimmune
	Fitzgerald P, Ndiaye NC, Visvikis-Siest S. Associations of vascular endothelial growth thyroid diseases. Autoimmunity. 2016 Aug 5:1-7. * equal first authors
	factor (VEGF) with adhesion and inflammation molecules in a healthy population.
	Cytokine. 2013. 61: 602-607. *equal first authors The aim of the thesis of Abdelsalam Saleh was to investigate the associations between the 6. Ndiaye NC*, Azimi Nehzad M*, El Shamieh S*, Stathopoulou MG*, Visvikis-Siest S*. VEGF related variants and its expression isoforms with its plasma levels and with cell Cardiovascular diseases and genome-wide association studies. Clin Chim Acta. 2011. adhesion molecules, inflammatory molecules, plasma lipids, candidate genes (NOS3, CD14, 412(19-20): 1697-701. *equal contribution MMP3 and IL-4) and with cardiovascular risk factors (obesity and blood pressure) in healthy
	individuals. In the same period, I co-directed the thesis of Marc Rancier : « Intérêt de la génomique du The publications related to this thesis are: VEGF en endocrinologie clinique » and Abdelsalam Saleh : « VEGF: Un biomarqueur potentiel dans la physiopathologie cardiovasculaire » which were presented on March 2015 1. Stathopoulou MG*, Bonnefond A*, Ndiaye NC*, Azimi-Nezhad M, El Shamieh S, Saleh
	(doctoral students 4 and 5). Their results and my implication will be detailed in the next
	section (Research Engineer Inserm).
	3. Azimi-Nezhad M, Herbeth B, Siest G, Dade S, Ndiaye NC, Esmaily H, Hosseini
	SGhayour-Mobarhan M, Visvikis-Siest S. High Prevalence of Metabolic Syndrome in Iran in Comparison with France: What Are the Components That Explain This ? Metab Syndr Relat Disord. 2012. Jan 27; 10(3): 181-8 4. Stathopoulou MG*, Bonnefond A*, Ndiaye NC*, Azimi-Nezhad M, El Shamieh S, Saleh A, Rancier M, Siest G, Lamont J, Fitzgerald P, Visvikis-Siest S. A common variant The aim of this report was to assess the epistatic interactions that are implicated in the Cytokine. 2013. 61: 602-607. *equal first authors determination of telomere length of children through GWAS and candidate gene approaches. 3. Zaaber I*, Rancier M*, Stathopoulou MG, Saleh A, Marmouch H, Masson C, Murray H, One publication was produced by this work: Kurth MJ, Lamont J, Fitzgerald P, Mahjoub S, Said K, Bel Hadj Jrad Tensaout B, Mestiri S, Visvikis-Siest S. Plasma VEGF-related polymorphisms are implied in autoimmune 1. Stathopoulou MG, Petrelis, AM, Buxton JL, Froguel P, Blakemore AIF, Visvikis-Siest S.
	highly associated with plasma VEGFA levels also contributes to the variation	of
	both LDL-C and HDL-C. J Lipid Res. 2013; 54(2): 535-41. *equal first authors

The aim of the thesis of Marc Rancier was to identify the VEGF related polymorphisms and the VEGF expression isoforms that could be useful for personalised medicine. Thus, we have studied the effect of the identified VEGF determinants on diseases intermediate phenotypes such as adhesion, and inflammation molecules, metabolic syndrome and lipid, while studies in auto-immune thyroid diseases were performed aiming to investigate links with thyroid hormones and thyroiditis.

The publications related to this thesis are: 1. Stathopoulou MG*, Bonnefond A*, Ndiaye NC*, Azimi-Nezhad M, El Shamieh S, Saleh A, Rancier M, Siest G, Lamont J, Fitzgerald P, Visvikis-Siest S. A common variant highly associated with plasma VEGFA levels also contributes to the variation of both LDL-C and HDL-C. J Lipid Res. 2013; 54(2): 535-41. *equal first authors 2. Azimi-Nezhad M*, Stathopoulou MG*, Bonnefond A, Rancier M, Saleh A, Lamont J, Fitzgerald P, Ndiaye NC, Visvikis-Siest S. Associations of vascular endothelial growth factor (VEGF) with adhesion and inflammation molecules in a healthy population. thyroid diseases. Autoimmunity 2016 Jun;49(4):229-35. * equal first authors 4. Rancier M*, Zaaber I*, Stathopoulou MG, Chatelin J, Saleh A, Marmouch H, El Shamieh S, Masson C, Murray H, Lamont J, Fitzgerald P, Mahjoub S, Said K, Tensaout BB, A, Rancier M, Siest G, Lamont J, Fitzgerald P, Visvikis-Siest S. A common variant highly associated with plasma VEGFA levels also contributes to the variation of both LDL-C and HDL-C. J Lipid Res. 2013; 54(2): 535-41. *equal first authors 2. Azimi-Nezhad M*, Stathopoulou MG*, Bonnefond A, Rancier M, Saleh A, Lamont J, Fitzgerald P, Ndiaye NC, Visvikis-Siest S. Associations of vascular endothelial growth factor (VEGF) with adhesion and inflammation molecules in a healthy population. Cytokine. 2013. 61: 602-607. *equal first authors 3. Saleh A*, Stathopoulou MG*, Dadé S, Ndiaye NC, Azimi-Nezhad M, Murray H, Masson C, Lamont J, Fitzgerald P, Visvikis-Siest S. Angiogenesis related genes NOS3, CD14, MMP3 and IL4R are associated to VEGF gene expression and circulating levels in healthy adults. BMC Med Genet. 2015;16:90. * equal first authors Also, I co-supervised the master II thesis of Alexandros Petrelis : « Investigations biostatistiques/bioinformatiques des interactions gène x gène impliquées dans le risque cardiovasculaire -la longueur des télomères leucocytaires chez les enfants » (master student 1). Genetic Determinants of Leucocyte Telomere Length in Children: a Neglected and Challenging Field. Paediatr Perinat Epidemiol 2015;29(2):146-150

Effects of LSR Polymorphisms on lipidemia and Alzheimer's disease (AD)

  ). Methylation detected in promoter regions of TPX2 and HAS-1 genes could explain previously reported associations with VEGF-A gene. In the second study, we have assessed associations between VEGF levels My involvement included the conception and design of the studies, the analysis and interpretation of the results and the reviewing of the manuscript. To implement the research results into clinical practice and establish the role of VEGF as a

	and inflammation molecules (IL1α, IL1β, IL2, IL4, IL6, IL8, IL10, INF-γ, TNF-α, MCP-1
	and epidermal growth factor (EGF)) levels measured in PBMCs extracts. Associations with
	VEGF gene expression isoforms (VEGF-121, VEGF-145, VEGF-165, VEGF-189) were also
	tested. We have identified significant associations between VEGF levels and levels of IL4
	(β=0.028, P=0.013), MCP-1 (β=0.015, P<0.0001) and EGF (β=0.017, P<0.0001).
	Furthermore, mRNA isoform VEGF-165 was associated with MCP-1 and IL1α (P=0.002 and
	P=0.008, respectively); and mRNA isoform VEGF-189 was associated with IL4 and IL6
	(P=0.019 and P=0.034, respectively) levels. This study provided insights of mechanisms
	implicated in VEGF regulation in PBMCs. The related manuscripts have been submitted for
	publication. predictive, preventive, diagnostic and prognostic biomarker
	 To provide information on the effect of VEGF "-omics" profiling in side effects and
	response to therapy through pharmacogenomics studies
	 To propose implementation strategies and European guidelines involving VEGF "-omics"
	profiling for the management of non-communicable diseases

This extended research on VEGF has led in the development of the Vascular endothelial growth factor European Genomic Federation Consortium (VEGF Consortium) by Sofia Siest as its leader and by me as a project manager (www.vegfconsortium.org) in 2015. The consortium aims to develop a transnational collaborative network dedicated to large integrative and multidisciplinary genomic studies of the VEGF in order to generate applicable knowledge for medical practice. Its objectives are:  To combine data from multiple cohorts in order to identify VEGF "-omics" profiling in health and non-communicable diseases  To elucidate the pivotal role of VEGF in the pathophysiology of non-communicable diseases  To demonstrate the patients' stratification potential of VEGF "-omics" profiling   To share methodologies, data and knowledge in the field of "-omics" management and innovative statistics Project 2 :

  Studies have focused particularly on ApoE's role in β-amyloid peptide (Aβ) deposition and clearance in the brain 106 , but little information is available on peripheral effects of ApoE isoforms on cognitive decline and AD. The ability of plasma ApoE to restore synaptic function despite lack of ApoE expression in the brain was recently reported 107 , suggesting a potential link between ApoE-dependent regulation of peripheral lipid homeostasis, cognitive function and AD. The lipolysis stimulated lipoprotein receptor (LSR) is involved in the regulation of lipid distribution between different tissues. It recognizes ApoB and ApoE and plays an important role in the elimination of triglycerides-rich lipoproteins during the postprandial phase 108, 109 .

	not been conclusive, or have even shown contradictory results 104 . This may be due to the
	complexity of dyslipidemias which result from perturbations in lipid homeostasis resulting in
	hypercholesterolemia, hypertriglyceridemia, or both (mixed hyperlipidemia) with changes in
	lipoprotein profiles that are not taken into account in the analyses. Studies evaluating efficacy
	of cholesterol lowering treatments such as statins to treat AD have proven inconclusive,
	suggesting that hypercholesterolemia is not the only risk factor involved 105 . It is therefore
	necessary to identify the key pathways regulating peripheral lipid homeostasis that influence

100, 101

. This paradox is further complicated by the fact that even if the clearance of dietary lipids in type III HLP patients is less effective in individuals with the APOE ε2 / 2 genotype, only a few patients ε2 / 2 actually develop the disease 102 , indicating that environmental factors and other genetic factors must contribute to Type III HLP 103 . Furthermore, in vivo studies have pointed towards potential links between dyslipidemias, agerelated cognitive decline and Alzheimer's disease (AD)

104

. However, results from epidemiological studies comparing lipid levels in controls and AD or dementia patients have risk for cognitive decline and AD. ApoE is the strongest risk factor for the sporadic form of AD, which represents > 95% of AD cases. The ε4 variant is a strong AD risk factor 101 and ε2 may lower the risk of AD and age of onset 106 , but the involved mechanisms remain as yet unclear.

Acknowledgments

I would like to express my deepest gratitude to Dr Sofia Siest for her support and guidance throughout my research career and for honoring me with her trust.

Next, I would like to thank the members of the committee that have accepted to evaluate this work for their time and their comments. I need to express my warm feelings towards my current and past colleagues in the IGE-PCV; U1122 unit, who have been my companions throughout the last 8 years of my life in France: Pr. Gerard Siest, Brigitte, Christine, Georges, Sebastien, Patricia, Alexandros, Vesna, Ting, Alan, Said, Abdel, Mohsen, Marc, Jerome, Payman, Coumba, Daniel, Bernard, and Michèle for helping me build my professional ambitions and for bringing joy every day at work. I also want to thank the supporting stuff and services of this unit, including the personnel of the University of Lorraine of the Lionnois site and the Inserm Grand Est, for ameliorating my work conditions. I would not forget of course to thank Prof. George Dedoussis, who was my first academic mentor, for his support and collaboration since 2003.

I would also like to acknowledge the collaborators of the different research projects that the IGE-PCV; U1122 unit and I personally am implicated, in regional, national and international level, for their trust, support and opportunities, including all the members of the VEGF Consortium.

Finally, my gratitude goes to my family: my husband Alexandros, my parents and my siblings for always believing in me, for their constant support, their patience and their unconditional love, but also to my friends in Greece and in France for always being there for me.

Organisation of conferences

I have participated in the organisation of the following conferences as members of the organising committee:  7th Santorini Conference Biologie Prospective, Santorini, Greece, 2014.

 Second ESPT Conference Pharmacogenomics: From Cell to Clinic, Lisbon, Portugal,

2013.

 6th Santorini Conference Biologie Prospective, Santorini, Greece, 2012.

My tasks included the communication of the conferences, the organisation of poster sessions, on site organisation and the drafting of the post-conference scientific manuscripts. In the 7th Santorini Conference Biologie Prospective, I was also member of the Poster Committee that was responsible for the evaluation of posters and selection of award winners.

Two more studies on VEGF "-omics" determinants are ongoing and are part of the thesis of Vesna Gorenjak, which I co-supervise (doctoral student 1). The first study aims to assess the epigenetic determinants of VEGF levels in 201 healthy individuals of the SFS through a discovery only epigenome-wide association analysis (EWAS). Twenty CpG sites within 13 I have drafted the manuscript that describes the development and the objectives of the Consortium [START_REF] Stathopoulou | A transnational collaborative network dedicated to the study and applications of the vascular endothelial growth factor-a in medical practice: The vegf consortium[END_REF] (original publication 7). This work was also presented in 2 international conferences as poster presentations (conferences 5 and 7). Three of the doctoral students cosupervised by me are involved in the development and the projects of the VEGF Consortium: Ting Xie, Jerome Chatelin and Vesna Gorenjak (doctoral students 3, 2 and 1 respectively).

I have also drafted two proposals entitled "The VEGF Project" on behalf of the VEGF Consortium that were submitted under the calls H2020-SC1-PM-02-2017: New concepts in patient stratification in 2016 and H2020-SC1-BHC-25-2019: Demonstration pilots for implementation of personalised medicine in healthcare in 2018. The first project did not receive the funding, while the second project is under revision by the European Commission.

Within the frames of the consortium, we have designed two projects that are ongoing and make part of the thesis of Jerome Chatelin, whose thesis I co-supervise (doctoral student 2).

One project is focused on the assessment of the role of the 4 VEGF-related SNPs (rs6921438, rs10738760, rs6993770, rs4416670) with BMD, age of menopause and risk of osteoporosis in a sample of 3 populations of post-menopausal women (OPG n= 52, SBCE n= 91, ARTEOS n= 97). We have identified a significant interaction between rs6921438 x rs6993770 and increased femoral neck BMD (b= 0.478, P= 1.52x10 -7 ).

In a second study we are assessing the effect of the 10 VEGF-related SNPs with levels of thyroid hormones and risk of hyperthyroidism and hypothyroidism in LifeLines cohort (LLs) (n= 4826), in Cilento Study (n= 2100), in CHRIS study (n= 3850) and in MICROS study (n= 873). Analysis results are currently being completed for the last two studies and a metaanalysis will reveal the significant results.

In these 2 studies, I am involved in the conception and design, in the selection of population and data preparation, in the analysis and interpretation of the results.

My participation in these projects has led to the design of a project on childhood inflammation within the frames of the CIWG consortium. Inflammation in childhood has been mostly linked with adiposity, obesity and obesity-related CVD 144 . In children, CRP levels have been associated with CVD risk factors, such as adiposity, obesity, and blood pressure, even in very young ages and these associations are most probably mediated by abdominal obesity (measured with waist circumference) 145,146 . Furthermore, there are indications that CRP is independently associated with advanced atherosclerosis in youth 147 and that CRP levels in childhood predict their values in adulthood 148 . Apart from CRP, TNF-α and interleukin 6 (IL6) have been associated to adiposity, visceral fat, insulin resistance and other CVD risk factors. 149 The identification of genetic determinants of inflammation markers in adults and in children could contribute significantly in the discovery of new genetic biomarkers of CVD risk and also in the identification of new biological pathways implicated in these associations.

The published GWAS are mainly focused on CRP while TNF-α and IL6 are not well represented [150][151][152][153][154][155][156][157][158][159][160][161][162][163][164][165] . Furthermore, the studies have been performed in adult populations (> 18 years old), thus excluding the investigation of children-specific genetic determinants.

Additionally, given the strong correlations between inflammation markers and adiposity, the use of this factor as a covariate in the regression models and the study of genes x adiposity interactions are likely to reveal genetic determinants that their effect could be masked by these interactions and which have not been previously discovered in GWAS.

I have proposed to lead a new project focused on inflammation in childhood within the CIWG. This project aims to identify novel childhood-specific genetic variants for CRP, TNFα, IL6 and other traits (such as white blood cells count) and to assess their interactions with abdominal obesity. GWAS on discovery and replication cohorts of healthy children (<18 years old), followed by meta-analysis will be performed. The proposal has been discussed and approved by the partners of the consortium, the analysis plan design has been circulated and the discovery populations' selection is ongoing.

Other projects

My participation in the project "Omics" data of SFS Project (described in the previous section Post-doctoral researcher), in the databases development and statistical analysis of the produced data has resulted in my inclusion in 3 original publications [166][167][168] (original publications 23, 24 and 20). These were large collaborative studies, where a large number of

Projet de Recherche: L'étude des déterminants « -omiques » du VEGF dans la médecine personnalisée des maladies chroniques

Scientific Project: The study of « -omics » determinants of VEGF in the personalised medicine of chronic diseases

Expected results

The major expected results are:

•

VEGF "-omics" determinants to be used for molecular make-up of CVD, AD and osteoporosis.

• Novel VEGF "-omics"-based risk prediction modelsalgorithms for CVD, AD and osteoporosis.

These results will have an important impact in the knowledge of VEGF biology and its implication in the assessed diseases, but also, will demonstrate its utility as a biomarker for personalised medicine through early risk prediction.

The knowledge generated by the project will be made available to the scientific community and the public following several publications in high impact factor international peerreviewed journals in the medical, genetics and epidemiology fields and with presentations in scientific conferences.

The VEGF consortium website will be used for the communication of the results within the consortium through a private area and with the general public through a public interface.

Summary of valorisation and transfer activities and scientific visibility

Total number of publications (peer-review journals): 66 The figure 8 demonstrates the numbers of publications by year.