Laura Calvillo, Cardiovascular condition like hypertension needs preclinical models able to discriminate among consequences of mechanical blood pressure increase (i.e. shear-stress) and biochemical action of ANG II, in order to test new therapies in a way that allows to distinguish more specifically their mechanisms of action, thus improving the evaluation of efficacy. Animal model is too complex and classic in-vitro cell culture too simple, therefore, advanced in vitro tools, able to better reproduce physiological conditions without the confounding effects of the various organs of the organism, are needed (Marchesi et al. Plos One 2020 15, 1–16.; Vozzi et al Biotechnol. Bioeng. 2011, 108, 2129–2140). The recent development of the IVTech Bioreactors-LivePa System, which will be employed in this project, provides for the first time an advanced model of hypertension without the need of animals. This system allows to compare the LivePa-dependent shear-stress effect with the biochemical action of ANG II on endothelial cells and to test potential protective effect of Imidazo-Pyrazolil Ureas (IPUs), (Selvatici et al European J. Pharmacology 718 (2013)428–434) (Brullo et al. Eur. J. Med. Chem., 47, (2012) 573-579). Therefore, the aim of this study is to stimulate HUVEC with LivePa or with ANG II, to separately mimic the two components of hypertensive stimuli, and then to treat them with the IPU selected among those able to inhibit hypertension-related inflammatory factors.

To this aim, HUVEC will be cultured in Bioreactor for 48 hours without flow to reach the confluence. Then, the flow will be started and cells will be treated with 1000 nM concentration of ANG II with or without IPU, vs LivePa-0.5 pressure increase with or without IPU, vs ANG II+LivePa with or without IPU. Appropriate positive and negative controls will be used. The optimal flow for HUVEC and the LivePa pressure parameters were set in our laboratory according to computational fluid dynamic model and device instructions. The following parameters will be assessed: morphological observation, viability test, p38 MAPK, NF-kB and hypertension-related cytokines production, which are factors playing an important role in hypertension (Calvillo et al. Nat Rev Cardiol 2019;16(8):476-490) (Dai et al. Sci Rep 
[Internet]. 2016 Sep 10;6(1):27600). If IPU will have protective effects on cells through our innovative in vitro model, it will be possible to hypothesize their employment in new therapeutic applications for hypertension. 

Mathieu Vinken, Several concerns have been raised in recent years regarding the safety of food additives. Among those are a number of colorants, sweeteners, anti-caking or glazing agents that have been shown to cause adverse effects in the liver of laboratory animals. The present project elucidates the mechanisms underlying these presumed hepatotoxic effects by incubating these food additives in tridimensional spheroid cultures of primary human hepatocytes and non-parenchymal liver cells. Through whole transcriptome templated oligo assay with sequencing read-out analysis and pathway analysis, a mechanistic scenario of the hepatotoxic effects induced by the food additives is established. This is substantiated by a series of translational and functional analyses of known triggers and key events in liver toxicity, in particular steatotic and cholestatic insults. Furthermore, the use of effects at the transcriptional level for setting limits for safe daily intake of food additives is explored. Overall, the outcome of the present project sheds more light onto the hepatotoxic potential of food additives and demonstrates the power of human-based in vitro experimentation for chemical risk assessment purposes.