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Technical viability of a novel device for automated high-volume screening of teratogens based on simultaneous quantitative computer-assisted microscopical evaluation of clonal cell cycle progression and differentiation state

Objectif



The developmental toxicity studies required by the regulatory authorities for registration of new pharmaceuticals and agrochemicals require a large number of animals and considerable effort from skilled technical personnel. The cost of testing a single chemical substance are approximately 250kECU, a factor which severely limits the number of novel agents and process derived by-products which can be evaluated. As a consequence the large number of novel compounds produced in research laboratories and those intermediates of step-wise procedures in the chemical industry, will never be tested by these in vivo procedures. In vitro methods provide an alterative strategy for assessment of developmental toxicity however the most promising methods are time consuming, involve animal use and require skilled personnel. Development of a simple, reliable and rapid screening system is required to facilitate the pharmaceutical and agro-chemical industries in evaluating embryotoxic potential in the large number of existing substances and the new materials which are added to the list each year.
EU Directive 86/609 dictates that animals must not be used for testing if a validated, non-animal test is available and the EU BIOTECH Programme has as a goal the development of such assays for the in-vitro detection of teratogens. Previously, members of this consortium have been contracted under the BIOTECH programme to identify in-vitro endpoints for the development of such tests (BIO2CT930471). As the most significant initial steps in the emergence of developmental toxicity is inhibition of cell division coupled with a premature differentiation, coincident evaluation of these endpoints was found to reliably discriminate known teratogens from non-teratogens in structurally related agents using single cells in-vitro. To determine change in differentiation state change in cell morphology was determined using an automated, computer assisted image analysis system which was developed specifically to allow unbiased estimations in large numbers of single cells. The aim of the present proposal is to modify this system in a manner which allows high volume, coincident analysis of proliferation rate and differentiation state; to develop a prototypic system with standard operating procedures; and to demonstrate the validity of the system for routine use in the public and industrial sectors. The proposal fits well with area 7.1.1 of the BIOTECH Work Programme and is within the remit of a Demonstration Project.
The three partners in the consortium, a cell biologist, pharmacologist and teratologist have all in various ways been employed in research dedicated to in-vitro screening for a number of years. As they have worked together to a considerable extent in the past the present initiative of this European research team is unique and calculated to give the maximum advance in providing an automated in-vitro screening system. DK and IE will modify the existing image analysis system for the coincident detection of cell proliferation rate and morphology. DK will develop the necessary software and. design, provide and install the prototype teratogen screening system in the laboratory of the IE partner. DE will synthesise and purify teratogenic and non-teratogenic analogues and provide these in a coded fashion for the purpose of validating the installed prototype. IE will be the co-ordinator with the responsibility of ensuring optimal producer-user relationships and that the technical viability of the novel technology overcomes the concerns of 'barrier groups' and obtains the support of 'opportunity groups' through the participation of extended audiences. The assignment of tasks in the project reflects these various expertises of each partner.

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NATIONAL UNIVERSITY OF IRELAND, DUBLIN
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Adresse
Belfield
4 DUBLIN
Irlande

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