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ZF-TOOLS<br />
High-throughput Tools for<br />
Biomedical Screens in Zebrafi sh<br />
Summary<br />
The zebrafi sh embryo model has rapidly gained ground<br />
in biomedical research based on functional genomics<br />
approaches. As a result it shows great potential to be incorporated<br />
into preclinical drug screening pipelines. ZF-TOOLS<br />
aims to develop a case study for an anti-tumor drug screening<br />
system, based on implantation of fl uorescently labeled<br />
tumor cells into zebrafi sh embryos. The optically transparent<br />
embryos provide a highly useful in vivo system to<br />
monitor growth and metastasis of implanted tumor cells.<br />
This system will allow for the powerful combination of visual<br />
monitoring with high-throughput analysis of expression<br />
of marker genes with a predictive value for tumor progression<br />
or for defence responses towards developing tumors.<br />
However, the bottleneck for application of this approach is<br />
insuffi cient knowledge of relevant disease marker genes in<br />
zebrafi sh. Therefore, the fi rst objective of ZF-TOOLS is<br />
marker discovery. To achieve this objective fl uorescent<br />
zebrafi sh cell lines expressing selected oncogenes will be<br />
generated and a multidisciplinary functional genomics<br />
approach, integrating diff erent genomics techniques and<br />
bioinformatics, will be undertaken to compare expression<br />
profi les before and after implantation into zebrafi sh<br />
embryos. The experimental design of our genomics strategy<br />
will enable distinction between tumor cell markers and<br />
markers for immune or general defence responses of the<br />
organism. After marker validation, high-throughput tools<br />
for large scale expression screens will be developed. The<br />
second objective of ZF-TOOLS is to exploit the combination<br />
of high-throughput markers and implant system to<br />
study diff erent oncogenic implants and eff ects of a test<br />
panel of drugs used in human cancer therapy. This case<br />
study should result in identifi cation of important tumor<br />
growth and metastasis factors as well as defence factors.<br />
Fundamental knowledge, tools and technical expertise<br />
gained from ZF-TOOLS will be commercially exploited by<br />
three high-tech SMEs.<br />
Problem<br />
Keywords | Zebrafi sh embryo model | functional genomics | tumor markers | defence response | cell transplantation |<br />
The ZF-TOOLS objectives are focussed on the incorporation<br />
of the zebrafi sh embryo model into the preclinical drug<br />
screening pipelines. The use of mice for in vivo monitoring of<br />
disease processes such as tumorigenesis, metastasis and<br />
immune response to tumors is limited by costs and throughput<br />
level. Introducing a high-throughput zebrafi sh embryo<br />
model could potentially contribute to cost-eff ective and<br />
more effi cient methods in the anti-tumor drug discovery<br />
process. Acceleration of drug lead time benefi ts economy<br />
as well as quality of life of cancer patients.<br />
The lack of basic knowledge of disease marker genes is<br />
the current bottleneck for biomedical research in zebrafi sh<br />
and for genomics-based compound screens in this model<br />
organism. The ZF-TOOLS project uses multidisciplinary<br />
functional genomics approaches to discover novel disease<br />
markers. The expected identifi cation of factors important<br />
for tumor growth and metastasis and organismal defence<br />
responses will contribute fundamental knowledge relevant<br />
to human health and will open the door to the<br />
establishment of zebrafi sh-based biomedical research<br />
and screening tools.<br />
Aim<br />
The ZF-TOOLS project is a coordinated eff ort of three<br />
research laboratories and three SMEs aimed at the following<br />
objectives:<br />
• genomic-based marker discovery for biomedical screens<br />
in zebrafi sh;<br />
• use of high-throughput marker analysis and tumor cell<br />
implants for the identifi cation of tumor growth and<br />
metastasis factors and organismal defence factors.<br />
The project aims to develop a case study for an anti-tumor<br />
drug screening system, based on the implantation of fl uorescently<br />
labeled tumor cells into zebrafi sh embryos. This<br />
innovative tumor cell implantation system is currently being<br />
developed by one of the SME partners in the project and has<br />
the major advantage that it does not involve the use of transgenic<br />
animals. Growth and metastasis properties of implanted<br />
tumor cells can be effi ciently monitored by fl uorescence<br />
microscopy during development of the transparent zebrafi sh<br />
embryos. The system resembles the natural situation of<br />
tumor growth, as the tumor cells are derived from zebrafi sh<br />
cell cultures of embryonic origin and implanted back into<br />
zebrafi sh embryos. We envisage that a powerful screening<br />
system can arise from the combination of high-throughput<br />
marker analysis with the possibility to visualize tumor growth<br />
and metastasis in an optically transparent vertebrate model<br />
organism. However, for realization of such a screening<br />
system, the identifi cation of relevant disease marker genes in<br />
zebrafi sh forms a crucial step. In the ZF-TOOLS project<br />
diff erent genomics approaches will be used to discover<br />
novel markers, which will be suitable for application in the<br />
ZF-TOOLS tumor screening system and also will have<br />
a broader utility for disease research in the zebrafi sh model.<br />
The experimental design of our genomics approach is<br />
expected to result in the identifi cation of two classes of<br />
markers:<br />
• markers correlating with growth and metastasis of tumor<br />
cells;<br />
246 CANCER RESEARCH PROJECTS FUNDED UNDER THE SIXTH FRAMEWORK PROGRAMME