• Principal Investigator: Dr Peter Kille – Cardiff School of Biosciences
• Collaborators: TIME, School of Medicine
  Prof Stuart Woodhead – Invitron Ltd, Wyastone Business Park, Monmouth, NP25 3SR
  Prof Keith Smith – Catcelt Ltd, 46 Radyr Avenue, Swansea, SA3 5DT
• Staff: Dr Ceri Morris
  Dr Gamal El-Hiti
• Funder: Welsh Government

Abstract

The primary objective of this project is to exploit Cardiff University’s unique position by combining its patented technology of genetic biosensors using “chemiluminescence” with its expertise in toxicogenomics to develop a portfolio of data demonstrating the feasibility of robust and rapid “animal-free” toxicology tests. Exploitation of this resource in the future will lead to the replacement of animal tests by establishing the transfer of known biological endpoints, routinely measured in in vivo animal trials, to more easily measured in vitro molecular endpoints within ‘animal-free’ tests.

Initial studies have demonstrated the feasibility of this approach. Preliminary data have demonstrated the high sensitivity and specificity of chemiluminescence-based assays of gene transcription products. Funding is required to achieve proof-of-concept through the development of a test system based on accepted genetic endpoints.

To convert the stated objective, the project aims to advance and exploit a platform technology that allows toxicological tests to be performed in a cell based, high-throughput format by quantifying genetic endpoints using an established chemiluminescence detection system. A modular platform is envisaged which may be configured to exploit any appropriate cell line together with its inherent molecular toxicological indices. Technical development will focus on optimising a homogeneous assay format comprising the following steps: cell exposure, cell lysis, RNA extraction, amplification (if required) and detection. The major challenges will be preserving assay sensitivity, specificity and precision within a high throughput format. The initial phase of the project will develop molecular endpoints for steroidogenesis within the immortalised adrenal cortex cells H295R, these will include genes such as those encoding cyp19, cyp21 and 3β-hsd2. This cell line has been identified by the US-EPA Endocrine Disruptor Screening Program (EDSP) as the in vitro tier 1 screen for substances that interfere with steroidogenesis. Its application in this context is presently progressing through validation, prior to scientific review and final regulatory acceptance and implementation. The successful development of this target system will enable feasibility to be established in an application that is already progressing through the process of regulatory acceptance. There are several other areas that are not yet adopted within the regulatory framework which will benefit significantly from the implementation of this technology platform. One such application is that of identifying chemicals that have genotoxic side effects (eg carcinogenesis). This application, which has high commercial potential particularly in the (non-regulated) screening of putative pharmaceutical agents, will also be explored within the project.