Cancer Research - Europa

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