Replacing animal experiments by direct measurement of DNA damage responses in human tumor slices ex vivo
Projectomschrijving
Ronde 2012 Module Proefdiervrije Technieken:
Ontwikkeling van antikankertherapieën vraagt veel proefdierexperimenten. In dit project hebben we nieuwe technieken ontwikkeld om menselijk tumormateriaal direct te kunnen gebruiken voor het uittesten van nieuwe medicijnen en behandelingen. Hiervoor hebben we de kweekcondities buiten het lichaam geoptimaliseerd en het tumormateriaal behandeld met het nieuwe medicijn (in dit geval een radioactieve stof). De tumoren verkregen direct na operatie van de patiënt zijn niet in muizen doorgekweekt, zoals vaak gebeurt, maar plakjes van deze tumoren zijn gekweekt in een kweekmedium waarin direct het effect van de behandeling getest werd. We hebben laten zien dat het mogelijk is om de tumorplakjes 5 dagen door te kweken en in die tijd verschillende therapieën te testen. Mede met deze methode hebben we een vernieuwde therapie ontwikkeld die mogelijk gaat leiden tot verbeterde patiënten behandeling.
Verslagen
Samenvatting van de aanvraag
Cancer research relies heavily on animal experiments. Both murine tumor models and xenografts in mice are commonly used for drug testing and fundamental oncological research. These models have been indispensable tools for our insight into tumor biology, drug sensitivity and preclinical evaluation of tumor response to therapy. However, the number of animals required for these experiments is substantial and some parameters cannot be translated easily from mouse to man. Therefore, we propose to develop novel technology employing primary human tumor slices ex vivo to test sensitivity to various (DNA-damaging) treatments as well as several basic parameters to increase our understanding of tumor responses to therapy. Tumor slices will be evaluated directly after collection of tumor specimens from patients and tested for responses to DNA-damaging treatments. We will concentrate on neuroendocrine tumors, which are generally treated with radiolabeled somatostatin analogs (which create DNA damage in cells that express the corresponding receptor). In addition to direct use of tumor material after collection from the patient, we will also start to develop methodology to freeze tumor slices in such a way that they can be thawed afterwards without significant loss of viability. Various aspects of the technology developed in this project will also contribute to introduction of similar ex vivo approaches in other areas of biomedical research that currently utilize mouse xenograft models. We expect that this strategy will reduce the use of mouse tumor models as well as xenografts, because several parameters can be measured equally well (such as apoptosis and several DNA damage response parameters) or even better (DNA repair kinetics, precise determination of drug distribution in the various cell types of the tumor, as well as in extracellular matrix and surrounding normal tissue) in ex vivo DNA damage response assays that utilize human tumor specimens.