Work Package 5: Small molecule screening

A major advantage of the zebrafish for drug screening is the possibility to test compounds at a medium to high throughput, as in a cell-based assay, but in the context of a whole organism, allowing to detect bystander effects that are not visible with in vitro assays. Thus zebrafish screens allow us to test for drug efficacy and toxicity in a single step.

We expect to identify a group of genes or disease-relevant processes as druggable targets based on their phenotypic analysis. Mutants of these genes or transgenic reporter lines for these processes will be subjected to small molecule screening to gain deeper insights into their regulation and function and to identify drug candidates.
For this purpose we will leverage robotic high-content, high-throughput screening methods and facilities being established at CNRS (France), KIT-G (Germany), BHAM and USFD (UK), as well as at ZFB (Spain), a company that focuses on drug discovery through zebrafish disease models. KIT-G and CNRS will use their expertise in informatics, computation, microfluidics and robotics to develop automated microscopes that can recognise the teratological or toxic effect of chemicals. They are also developing new microfluidics devices for robotic delivery of test drugs, and are developing algorithms to detect changes in fluorescence intensity and shape and animal movements. BHAM has developed robotic microscopy for quantifying fluorescent transgene expression. UL has developed high throughput screen systems for cancer and inflammation. ZFB has developed techniques that allow to obtain large quantities of zebrafish embryos, with high viability, synchronous development and without relying on natural spawning, as a basis for chemical screening.

KIT-G has a collaboration with the French Chemical Library Consortium and can access via this route about 30,000 chemicals, in addition to commercially available libraries. In parallel, it is building its own combinatorial synthesis facility to synthesise large quantities of specific chemicals, generate new libraries, and to synthesise structural variants of lead compounds. All these libraries will be shared between the consortium partners.

(Work Package Leader: KIT-G)

Dissemination of results

Hits from high-throughput screening will be confirmed by re-testing and by employing a large array of cell biological methods and biochemical methods available at the laboratories of the project partners involved in the screen. Structural variants of positive entities will be synthesised and tested to establish structure function relationships.

Compounds that give validated hits will be used as research tools for the analysis of regulatory pathways in development and disease (to be published mainly through articles in refereed scientific journals). If suitable they will also be licensed to pharmaceutical companies as a starting point for drug development.