Universiteit Leiden

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Proefschrift

High throughput microscopy of mechanism-based reporters in druginduced liver injury

Promotor: B. van de Water

Auteur
Steven Hiemstra
Datum
09 november 2016
Links
Thesis in Leiden Repository

Drug Induced liver injury (DILI) is a major problem in the clinic and a big economic treat to the pharmaceutical industry. DILI is often missed in preclinical stages of drug development as it occurs only in rare cases. Adaptive cellular stress responses are paramount in the healthy control of cell and tissue homeostasis and generally activated during toxicity in a chemical-specific manner. Only in susceptible individuals drug exposure can not only activate stress responses, but also switch on cell death signaling leading to liver injury. Therefore, monitoring adaptive stress response activation is key in prediction of DILI. In this thesis we established a platform containing a panel of distinct adaptive stress response reporter cell lines based on BAC-transgenomics GFP tagging in HepG2 cells. We validate and test these reporters in chapters 2 and 3. In chapter 4 we validate the system in a 3D environment where HepG2 cells show an enhanced liver like phenotype. In chapter 5 and 6 we show this reporter system can also be used for mechanistic research by unraveling crosstalk between DNA damage and Nrf2 signaling and by identifying novel regulators of Nrf2 signaling. Together, this thesis contributes to a more elaborate understanding of DILI.