Time resolved characterization of the development of NAFLD and the role of the gut and white adipose tissue therein.

Project description of the PhD trajectory Eveline Gart Authors: R.Kleemann (mentor at TNO), M. Morrison (daily supervision) and I. Bobeldijk-Pastorova (project manager ProLiver) Date: October 30th 2017 Embedding of project: The PhD trajectory of Eveline Gart is aligned with the strategic scientific work on: a) the development of non-alcoholic fatty liver disease (NAFLD) in obesity at TNO Metabolic Health Research (Leiden) and b) the work that will be performed in a recently established private public partnership (PPS ProLiver) which has been launched by TNO and partners (from nutritional industry, pharmaceutical industry, assay development SMEs). The PhD trajectory is aligned with the roadmaps of TNO’s LIFE cluster and the focus areas of the TopSectors LSH and Agro&Food from which funding is obtained for the PPS ProLiver. Aims: TNO Metabolic Health Research aims at obtaining insight into the development of NAFLD in order to establish translational models for R&D and a first therapeutic treatment for this disease. Furthermore, PPS ProLiver aims at defining key mechanisms of NAFLD development and assesses the role of adipose-tissue (obesity) and the gut (microbiota) to NAFLD progression. Background: NAFLD is the most prevalent form of chronic liver disease worldwide and there are no therapeutic treatments available. NAFLD has an estimated incidence of >20-35% in the general population and a particularly high prevalence in children with overweight (~ 80%). Because there are no therapies available to stop disease progression, NAFLD is becoming one of the greatest global challenges (unmet medical need). The PhD will investigate the development of NAFLD in specific risk phenotypes (obesity; dyslipidemia; insulin resistance) over time using translational models and omics technology in order to identify key disease mechanisms, functional biomarkers and to define new therapeutic approaches to treat the disease using (combinations of) nutrition and pharmaceutical therapies. To achieve this, ProLiver will apply new biomedical technologies and methods, a part of which were developed at TNO (strategic research in TNO’s early research programs). The PhD will conduct her own research using the infrastructure and resources provided by TNO. The PhD will also intensively collaborate with leaders in the field (e.g. human pathologists/AMC) and ProLiver partners. The PhD trajectory of Eveline Gart is a logical continuation of the long-lasting collaboration research collaboration between TNO-Metabolic Health Research (Dr. Martine Morrison, Dr. Robert Kleemann) and our partner at Wageningen University (Prof Dr Jaap Keijer, Dept of Human and Animal Physiology; promotor of PhD). The two groups collaborate for ~10 years successfully and this has resulted in several research publications and research projects (e.g. at Top institute food and nutrition; FP6 program; Nutrigenomics Organisation NUGO). Main project phases of the PhD trajectory: The project has three main phases, each with subprojects with publications. In phase 1 (~18 months), we will investigate the role of the gut and the adipose tissue in the development of NAFLD including microbiota analysis and recently established gut permeability function tests, as well as comprehensive bile acid and short chain fatty acid profiling (metabolomics). In addition, the contribution of inflammatory mediators released from adipose tissue will be assessed on basis of previous work (e.g. Mulder et al. Int J Obes, 2016; Schoemaker et al. PLoS ONE 2017; Mulder et al., Sci.Rep., 2017). In phase 2 (~18 months), we will compare NAFLD in different risk populations (obese subjects versus lean subjects; children vs adults) using translational models to identify unifying mechanisms as well as risk-group specific mechanisms allowing more stratified or personalized medicine. The models are well-established and represent specific NAFLD phenotypes or allow study of NAFLD in early life conditions (e.g. Wielinga et al., Mol Nutr Food Res., 2012; Morrison et al. J Hepatol., 2015; Arnoldussen et al., Int J Obes., 2017; Zimmer et al., Hepatol. Commun. 2017). Prototype conceptual interventions with nutrition and drugs will be performed. In phase 3 (which starts in parallel with phase 2 and continues until the end of the PhD trajectory), the obtained results will be disseminated together with partners and organizations relevant for patients (e.g. Dutch dietician education center, Dutch medical training centers at academic hospitals, MLDS) and at conferences or peer-reviewed publications. Independent reviewing of project: The scientific quality and applied character of the project has been examined independently by external referees of five companies, an independent commission of TNO (IBC committee), an independent ethical commission (DEC; for sections involving test animals); and the plans have been approved by the TopSectors LSH and Agri&Food. Scientific quality: The scientists (daily supervisor Dr.Morrison, mentor Dr. Kleemann, project leader Dr. Bobeldijk-Pastorova) are employees of TNO-Metabolic Health Research and their group has published >100 studies in the area of metabolic disease, obesity, NAFLD/NASH and the analytical technologies required for the project. The used preclinical models and technologies are well-established in the scientific literature with more than 250 high impact publications. Recent publications (2016 and 2017) include: 1. CAT-2003: a novel SREBP inhibitor reduces steatohepatitis, lipids and atherosclerosis in APOE*3-Leiden mice. Zimmer et al., Hepatology Commun., 2017 (in press) 2. A Casein Hydrolysate Based Nutritional Formulation Attenuates Obesity and associated NAFLD and Atherosclerosis in LDLr-/-.Leiden Mice. M.H. Schoemaker. PLoS ONE. 2017, 3. The CCR2 inhibitor propagermanium attenuates diet-induced insulin resistance, adipose tissue inflammation and non-alcoholic steatohepatitis. P.Mulder et al., PLoS ONE, 2017 4. Prolonged high fat diet exposure induces dynamic and adipose-depot specific DNA methylation in adult mice. R.A.J. Zwamborn et al. Sci. Reports, 2017 5. Intervention in white adipose tissue inflammation with rosiglitazone attenuates development of non-alcoholic fatty liver disease in obese LDLr-/- mice. P.C.A. Mulder, Scientific Reports, 2016 6. RvE1 attenuates atherosclerosis in absence of cholesterol-lowering effects and on top of atorvastatin. K. Salic, M.C. Morrison, L.Verschuren, P.Y.Wielinga, L.Wu, R.Kleemann, P.Gjorstrup and T.Kooistra. Atherosclerosis, 2016. 7. Thesis M.Morrison: Metabolic Inflammation in hepatic and vascular disorders. Strategies to attenuate disease development. ISBN 978-90-367-8669-0 (2016) 8. Intervention with a caspase-1 inhibitor reduces obesity-associated hyperinsulinemia, NASH and hepatic fibrosis in LDLR-/-.Leiden mice. M.C. Morrison, Int.J.Obes (London), 2016. 9. Surgical removal of inflamed epididymal white adipose tissue attenuates the development of non-alcoholic steatohepatitis in obesity. P. Mulder, M.C et al. Int. J. Obesity, 2016.