In the last years, a new concept of personalised medicine called Mouse Avatars' or co-clinical trials' has emerged. Development of mouse avatars implicates implantation of
patient tumour samples in mice for use in drug efficacy studies. This model allows conducting
preclinical trials in parallel with ongoing human phase I/II clinical trials. Murine and
patient trials are conducted concurrently, and information obtained from the murine system is
used for clinical management of the patient's tumour. The advantage of this approach is that
each patient has his/her own tumour growing in an in vivo system, thereby allowing the
identification of a personalised therapeutic approach. Such approach eliminates the cost and
toxicity associated with non-targeted chemotherapy. Unfortunately, nowadays, it is clear that
co-clinical trials using mouse avatars are not affordable because mice are expensive and
time-consuming. The study challenge is to make Avatars available for every patient and the
approach sustainable for the National Healthcare Systems. To implement this concept, the
investigators propose to replace mouse Avatars with zebrafish Avatars. Indeed, the
investigators propose to run co-clinical trials by using zebrafish embryos. Zebrafish embryos
as model for human cancer cell xenografts offer several advantages:
Rejection-free model. In zebrafish thymus is not working until 9 days post-fertilization
(dpf), allowing rejection-free xenografts during this period. Indeed, the use of
expensive immune-permissive strains is not required, in sharp contrast to the mouse
model.
Very low cost of husbanding.
Reduced amount of testing material. Zebrafish experiments require much less material to
assess drug efficacy.
Short time for the analysis of xenotransplants. The time required for a drug efficacy
study is 5 days, as opposite to several weeks or months in the mouse model.
Low experimental cost and simple procedures. The zebrafish produces large number of
embryos with each fecundation (hundreds). This provides statistical power to the
analysis and facilitates the collection of a large number of data.
Low ethical impact. Zebrafish is classified as insentient from fecundation to the time
at which embryos become capable of independent feeding (120 hours post fertilization,
hpf); therefore, during this time it does not require a license according to the
Directive 2010/63/EU.
Data collected in zebrafish are relevant to humans. Zebrafish genome is closely related
to that of humans. For instance, remarkable similarity in molecular signalling
processes, cellular structure, anatomy, and physiology has been observed between
zebrafish and other high-order vertebrates, including humans. This accounts for the
exponential increase in the use of zebrafish in drug discovery during the last two
decades.
Collectively, these points outline the remarkable advantages of the zebrafish Avatar over the
mouse Avatar, paving the way for a realistic and cost-sustainable implementation of the
co-clinical trials.
Specifically, the aim of the study is to perform an observational prospective clinical trial
on patients operated of epato-biliar-pancreatic cancers and gastro-intestinal cancers
undergoing a chemotherapy treatment in order to demonstrate that zebrafish Avatar is able to
predict the therapeutic regimen with the best efficacy for each patient. To this effect, 120
patients meeting the trial inclusion criteria will be enrolled over a 18 months period. In
this study, a fragment of tumor will be taken from the surgical specimen by the pathologist,
fragmented and transplanted in the yolk of 48 hpf zebrafish embryos. The effect of different
anticancer drugs and/or their combinations on the survival, proliferation and migration of
the xenotransplanted cancer cells will be evaluated by exposing the xenotransplanted embryos
to fish water modified with the drugs. The chemotherapy regimens to be tested in the
xenografted embryos are selected in agreement to the common clinical practice, i.e.:
fluoropyrimidines, platinum compounds, irinotecan, taxans (docetaxel) in stomach &
esophageal cancers
fluoropyrimidines, platinum compounds, irinotecan, gemcitabine, nab paclitaxel in
pancreatic/biliary cancers
fluoropyrimidines, platinum compounds, irinotecan in colorectal cancers.
Embryos will be analysed 4, 24 and 48 hours post injection (hpi). Primary measures will
include:
The enrolled patients after the surgical operation will undergo to the adjuvant chemotherapy
treatment.
The comparative evaluation undertaken after closure of intake will be based on prospectively
collected data on (i) clinical outcome and (ii) chemo-sensitivity data collected in zebrafish
model.