Axis 2 | Mechanical constraints on hepatocyte phenotype and genotoxicity

Dynamics of MicroEnvironment and Cancer [Dymec]

3D cultures, inside biomaterials are getting more and more developed, allowing an analysis of phenotypic variations, which are depending on exogenous factors and environmental contaminants such as heterocyclic aromatic amines. In the team, developed and optimized 3D models of cultures of normal and transformed liver cells (rat, mouse, human) in a context of various tension forces. These cultures may allow many applications in regenerative medicine and tissue engineering.

·
  • Implementation and optimization of a model of spheroid and organoid of human liver cells in 3D culture, for the purpose of cellular engineering.

The maintenance of the differentiated as well as biotransformation of xenobiotics functions make 3D hepatic cultures a model of choice for applications in biotechnology. We confirm the advantage of these cultures for the maintenance of the xenobiotic metabolism enzymes, in long term period, transformed cells derived from hepatocarcinomas and primary human hepatocytes (CRB Santé, CHU-Rennes, SFR Biosit)
We construct complex 3D structures of liver cells in matrices mimicking the in vivo architecture, and perform 3D prints of parenchymal and non-parenchymal cells (3D co-cultures of organoids) using a 3D printer, by micro-extrusion. This technology allows to carry out precise cellular arrangements, as well as a robotization with a reproducibility and precision of a few microns in three dimensions.

·

 

  • Genotoxic effects analysis of environmental contaminants

It is increasingly recognized that some environmental contaminants strongly influence the incidence of liver damage in humans and are thought to play a role in the incidence of liver cancer. 80% of human cancers are suspected to be due, in part, to environmental and dietary contaminants. It is difficult to extrapolate the results obtained from rodents to humans because of the strong differences in expressions and affinities of the biotransformation enzymes. In normal and transformed human hepatocytes cultured in 3D, we determine the metabolic activation of contaminants and identify DNA adducts in order to determine in humans the genotoxic potential of these compounds with acute or chronic treatments.
In recent years, our approaches have allow to understand the genotoxicity of Heterocyclic Aromatic Amines (contaminants of concern) in human liver by such in vitro approaches but also in silico (axis 4).

·