An Ex Vivo Model For The Assessment Of Drug Toxicity On The Human Retina

Abstract Purpose: Retinal toxicity is a common cause of drug development attrition. The aims of this research were therefore to develop the ex vivo human retina as a suitable model for the assessment of retinotoxicity, and to explore the methods in which this may be investigated. Methods: Post mortem and living donor human eyes were obtained, and the retinas dissected within 24h post mortem or 1h enucleation respectively. The ex vivo human retina was characterised using immunohistochemistry and QRT-PCR. The effect of multiple retintoxins was investigated on the human retinal cell lines MIO-M1 (Müller cells) and ARPE19 (RPE cells), and CHQ on the human organotypic retinal culture (HORC) using the LDH and MTS assays. TUNEL, Western blotting and QRT-PCR were also used to investigate the effect of CHQ on the HORC, and CDK expression investigated by QRT-PCR. Results: Cell specific markers were investigated in the post mortem and living donor, both possessed similar immunohistochemical and mRNA properties. CHQ was the most potent retinotoxin investigated in the cell lines, and when applied to the HORC, measureable toxicity was found along with an increase in the expression of multiple cell specific mRNA’s. The expression profile of multiple CDK’s in the ex vivo retina was investigated in relation to a retinotoxic Pan-CDK inhibitor, where differential expression was found. When exposed to the retinotoxic pan-CDK inhibitor, the cell lines displayed differences in toxicity. Conclusion: The ex vivo human retina is an ideal tissue to investigate retinotoxicity. It possesses similar properties as the in vivo human retina, and displayed measureable toxicity when exposed to CHQ. The ex vivo human retina also proved its usefulness in the investigation genes associated to a novel retinotoxin. The ex vivo human retina could act as a bridge between animal and human studies, providing vital information about a drug’s potential retinotoxicity.

Name dc:type.qualificationname

phd

Level dc:type.qualificationlevel

doctoral

Grantor dc:publisher.institution

University of East Anglia

Year dc:date.issued

2016

Author dc:creator

• Wright, Phillip

Language dc:language

en