Most people tend to focus on regulatory testing and the
need for properly validated in vitro tests to replace
regulatory prescribed animal testing when talking about
in vitro testing. In addition, the public tends to focus
on in vitro tests to replace animal testing for cosmetics
which, however, amounts to a very minor percentage
(smaller than 1%) of all animals used.
However, regulatory testing is only one side of the coin,
accounting for approximately
30-40% of animal testing in a country. Annually, millions
of animals are used in biological research programmes and
in compound/drug screening and development programmes.
With respect to in vitro testing, a clear distinction
should be made between (1) regulatory testing; (2)
biological research and (3) product development screening
and testing.
Over the past few years, the use
of animals for regulatory testing in Europe has declined
substantially. According to figures presented by country
representatives during the symposium 'Implementation of
the 3R targets in the EU, in science and in industry
(Vienna, Nov 16/17, 1998), animal testing is reduced:
* by 50% between 1995-1997 in Belgium;
* by 30% between 1987-1997 in Finland;
* by 30% between 1990-1997 in France;
* by 60% between 1991-1997 in Germany;
* by 65% between 1991-1997 in Austria;
* by 5.3% between 1997-1998 in the Netherlands (NCA
Newsletter 3/2000).
Although some EU member states continue to be reluctant
regarding the acquisition and inclusion of animal data in
these statististics, there is a clear indication that the
total number of animals used is indeed decreasing.
In toxicology/safety testing
departments of industry, awareness of animal testing and
alternatives is relatively high. For example, company
Bayer reported a decrease in animal testing of 85%
between 1984-1998 (Vienna, 1998), while company Novo is
regularly reporting a 50% decrease in number of animals
used for biological product control (Novo Nordisk,
Environmental & Social Report, 1999, see also Novo
website). Apart from adopting alternative tests where
approved by local or international regulatory
authorities, much can be gained with respect to the 3 R's
(replacement, reduction, refinement) by looking
critically at one's own approach to animal testing,
necessity of test, the way animal tests are performed,
the existence of alternatives etc. These actions may
result in substantial savings in numbers of animals used
and many refinements (Novo Nordisk Health Care, IVTIP
meeting 1999; Jansen Research Foundation, personal
communication).
In 1990 ECVAM (the European
Centre for the Validation of Alternatives to Animal
Methods, covering Europe) and ICCVAM (the USA Interagency
Coordinating Committee on the Validation of Alternative
Methods) were established. Over the years, various
prevalidation studies have been carried out by these
organizations. Within the EU, this has resulted in the
recent acceptance of three non-animal tests for topical
toxicity testing:
* a phototoxicity test (3T3 NRU test);
* two skin corrosivity tests (EpiStain Test,
TER-Transcutaneous Electrical Resistance test).
The EU guidelines for these three tests are sent to OECD
and will hopefully achieve world-wide acceptance soon via
the OECD member countries.
* the local lymph node assay (and 'ex vivo' test in mice
used for testing skin sensitization) has been endorsed by
the ECVAM Scientific Advisory Committee but still needs
formal acceptance by the competent authorities of the EU
member states and by OECD member countries. The test
significantly reduces the number of mice used for
testing.
While only few non-animal tests
have been validated and accepted world-wide, some
national authorities have anyway accepted the use of
alternative tests. With research on alternatives
progressing, industry as well as regulatory authorities
increasingly tend to accept in vitro tests that provide
reliable and consistent information on relevant
endpoints. Therefore, on a case by case basis, regulatory
acceptance of high quality, alternative tests performed
according to Good Laboratory Practice is expected to
increase, although these tests not necessarily have
undergone a full validation study.
(1) to promote the critical
in-house review of animal testing among its own member
companies and industry in general;
(2) to exchange information
about regulatory accepted alternatives to animal testing,
either fully validated or non-fully validated;
(3) to engage in a dialogue with
ECVAM/ICCVAM about the use of proprietary compounds in
validation studies;
(4) to promote the transfer of
the results of EU-funded projects on in vitro testing
alternatives to industry, in particular where it concerns
demonstration and prevalidation projects.
A substantial number of animals is
used in biological research and for human and veterinary
medicine studies: in some countries 4 times as high
(2,113,600) as the number used for safety evaluations
(543,200) (UK Statistics of Scientific procedures on
Living Animals, 1999). The past 10-12 years saw a decline
in numbers of animals used, but some countries, such as
the UK, currently report an increase in the number of
animals used for research. Much of this increase is
credited to the advent of transgenic animals for
biological studies: when making a line of transgenic
animals for research, those animals who are not
homozygous for the transgene will not be used for the
experimental procedure but are culled.
Among the academic research
community, awareness of alternatives to animal testing is
slowly increasing and is mostly seen in the field of
toxicology. Still, the academic research community is
adviced to be more aware of the use of in vitro tests to
answer fundamental biological questions. The quality of
research is proving to be very high indeed, as the
following information will demonstrate.
A survey conducted in 1986 has shown that more that 50 of
the 76 Nobel Prizes in Medicine or Physiology awarded
between 1901-1985 were given for studies in which
alternative methods made a major contribution to the
research for which the Prize was given. The prevalence of
alternative projects among the Nobel Prize winners
throughout the 20th century has increased steadily: from
1965 to 1985, 19 of the 20 prizes were for studies in the
'alternatives' category. This has resulted primarily from
the increasing prominence of in vitro studies, including
molecular and biochemical studies of subcellular
components, studies of cells and tissues in culture, and
mathematical approaches (Advances in Animal Welfare,
revised 1999).
The In Vitro Testing Technology
Transfer Project, an EU funded pilot study executed in
the year 2000, has shown that EU-funded projects on in
vitro testing have yielded many basic biological results
of high importance to industry, although these benefits
were often not the immediate objective of these
studies.
(1) to promote among their own
company members and academics in general the awareness of
alternatives to animal testing in biological
research;
(2) to urge the European
Commission that project proposals in the field of
biomedical research are evaluated separately on the
proposed use of animals in the research projects foreseen
and to award a higher score to those projects using
alternatives to animal testing;
(3) to urge the European
Commission to adopt a dedicated technology transfer
scheme to maximize the transfer of fundamental research
knowledge to industry;
(4) to urge the European
Commission to safeguard the continuity of the research
pipeline on in vitro testing, and to safeguard ample
funding for ECVAM for prevalidation and validation
studies;
(5) to urge the European
Commission to recognize, in its funding schemes, the
importance of highly innovative, high-risk and very long
term research that does not immediately seem to lead to
increased revenues or industrial competitiveness.
A wide variety of industries active
in the areas of cosmetic, chemical, pharmaceutical and
vaccine development use alternative, often in vitro tests
for lead compound screening and testing, and to determine
e.g. geno, hepato, neuro, nephrotoxicity etc. and
permeability/absorption.
The early stages of compound
development in the pharmaceutical industry represent
multiple opportunities for the use of in vitro assays,
which are typically miniaturized and/or automated to take
account of low compound availability and high
throughput.
As a first step, high throughput in vitro efficacy assays
may be used to screen chemical libraries to select a lead
compound, around which a large lead series is
synthesized. In a second step, medium throughput in vitro
assays (often cell-based assays) are used to screen these
compounds for their toxicological profile. A third step
often involves in vivo exposures to reveal organ specific
toxicity. If present, medium to low throughput screens
are used in a fourth step to select those compounds with
least target organ toxicity (e.g. primary hepatocytes for
hepatotoxicity, micromass or whole embryo assays for
teratogenicity).
Thus, the number of animal tests required during a
compound's development cycle has decreased substantially,
but there is a catch: because of the step-wise, more
controlled approach, the number of compounds under
development at a company is increasing, thereby
increasing demand for unavoidable in vivo tests. This
should be realized when evaluating statistics on the use
of animals in product screening and development.
Apart from a contribution to
reduced use of animals per compound under development,
alternative and in vitro tests offer an additional
advantage: they often give more objective answers than in
vivo tests. In vitro skin irritation models are, for
example, often more sensitive, specific and predictive of
the human situation that the existing in vivo skin
irritation models. Efforst to develop reliable in vitro
eye irritation tests are large and ongoing.
In total, therefore, the use of a series of in vitro
tests in industry contributes to better and faster
selection of candidate compounds, avoiding late
(negative) surprises in the product development
pipeline.
Basic research on in vitro testing therefore contributes
to competitiveness of European industry by providing
knowledge and tools for product development.
Research on in vitro testing
previously funded by the European Commission (Framework
Programmes 3, 4) focused on basic, enabling technologies
with applications both inside and outside the in vitro
testing community. Since few of the results generated
were adopted directly in industry or submitted to ECVAM
for (pre)validation, the value of these studies was
questioned.
The In Vitro Testing Technology
Transfer Project, conducted in 2000, has conclusively
shown that successive Framework Programmes definitely
yield a large amount of industrially important research
findings. However, it takes a special effort to identify
these results because of the lack of a coherent
technology transfer programme in the various Framework
Programmes.
Without a dedicated technology transfer initiative, it
will be difficult to place useful results in the
appropriate industrial context. In general, industry
finds it difficult to access the results of publicly
funded research, even when such results have been
presented, summarized etc. Again, the In Vitro Testing
Technology Tranefer Project has shown that the role of
independent consultants as pre-viewers of results is very
welcome.
(1) to increase among the own
member companies and industry in general awareness about
the use of in vitro and alternative approaches to
compound screening and development;
(2) to act as a clearing house
for results generated in publicly funded European
research projects in this field;
(3) to indicate to contractors
the potential industrial applicability of results, even
when results are not suitable for large-scale validation
programmes;
(4) to serve as a forum where
contractors can present their results and liaise with
industry; often, such liaisons result in industrial
contracts;
(5) to urge the European
Commission to put a proper technology transfer scheme in
place, preferably one that invites in depth reviews by
independent consultants on industrial applicability;
(6) to urge the European
Commission to allocate 5% of a future project's budget to
be allocated to technology transfer activities;
(7) to urge the European
Commission to prioritize independent reviews of in vitro
testing studies both in terms of money and in terms of
manpower.
IVTIP exists since December 1993
and has liased with the European Commission on in vitro
testing projects in FP 3 and FP 4. Commission officials
shared information with IVTIP about Commission
initiatives in the field, and this information was
distributed widely to IVTIP member companies.
Since 1993, IVTIP maintained close
contacts with the contractors of FP 3 and FP 4, sharing
information, inviting them to present results at plenary
meetings and liaiing at full sectoral meetings. Many of
the industry/academic contacts made during these meetings
resulted in joint projects and industrial contracts. On a
personal title, many representatives of IVTIP member
companies served the Commission as experts to evaluate
project proposals.
In earlier EC programmes and in FP
3 and FP 4, a separate (and increasing) budget was
available for in vitro testing projects: 1,94 million
EURO in the BAP programme, 4,5 million in the BRIDGE
programme, 8,67 million EURO in FP 3 and 15,25 million in
FP 4 (the latter two under the BIOTECH programme). A
separate programme line for in vitro projects was
available since 1992. When projects were evaluated for
funding, they only had to compete with projects
addressing comparable objectives. As a result, very good,
targeted projects were selected.
In FP 5, the situation with regard
to support for in vitro testing projects has deteriorated
in comparison with FP 3 and FP 4. In vitro projects are
covered under several different key actions, generic
projects and infrastructures. As a result, different
scientific officers should be liased with to obtain
information about the programme, which does not
facilitate the distribution of results from projects and
the identification of industrially important results.
Neither does it facilitate networking and the building up
of trust allowing for proper technology transfer
activities.
To make matters worse, a dedicated budget for in vitro
testing projects is lacking; in vitro projects have to
compete not with each other but with areas such as the
development of new drugs, gene therapy or DNA vaccines.
Because of their obvious human component, projects in
these areas traditionally score higher than projects
aimed at reducing animal experimentation.
After the 1st and 2nd call for
proposals, only few in vitro testing projects have been
selected for funding. This erodes confidence of academic
and industrial scientists working in the field in the
support of the European Commission for the development of
in vitro tests, and jeopardizes the future of the network
built up in the preceding years.
(1) to serve as a focal point
for the distribution of information from the European
Commission to member companies in particular and industry
in general;
(2) to serve as a focal point
for various scientific officers for the distribution of
information from individual projects to industry;
(3) to serve as an interactive
forum where contractors can present their results and
network one on one with industry;
(4) to stimulate contractors of
different but related projects to consult each other for
cross-fertilization and exchange of ideas and
experiences;
(5) to serve as a source of
industrial expertise for project evaluations and
technology transfer activities;
(6) to urge the European
Commission to award project proposals addressing
alternatives to animal testing extra points, as they
address a very urgent socio-economic need with important
ethical dimensions;
(7) to urge the European
Commission to improve administrative procedures regarding
the selection and funding of projects in such a way that
confidence among industry and academia in the EU-research
system is maintained.
Rotterdam, October 26, 2000