Determination of the genotoxic potencies of natural toxins using the γH2AX assay

Stage

Determination of the genotoxic potencies of natural toxins using the γH2AX assay

Recently, RIKILT has implemented the γH2AX assay for simultaneous detection of genotoxicity and cytotoxicity in the human liver cell line HepaRG. The test is based on the quantification of phosphorylation of histone H2AX (γH2AX), a process which is induced by double strand DNA-breaks, and is used to assess the genotoxic potential of compounds.

HepaRG is since it is known to be a highly metabolically active cell line and many natural toxins, particularly plant toxins, require metabolic activation to become genotoxic. Differentiated HepaRG cells express various cytochrome P450 and phase II enzymes at levels that resemble those found in primary human hepatocytes.

Aim

The aim of this project is to determine the relationship between concentration and genotoxic effect of various natural toxins using the γH2AX genotoxicity assay in the context of HepaRG cells. For that purpose,  HepaRG cells are treated with increasing concentrations of the toxins and subjected to the γH2AX assay. Subsequently, Bench Mark Dose (BMD) analysis of the γH2AX data is performed using publically available software. 

Supervisors: Geert Stoopen, Ad Peijnenburg 

If you are interested in this topics, send your application (letter and curriculum vitae) to Ad Peijnenburg.

Time schedule