Detect mitochondrial toxicity of novel therapeutics using Cyprotex’s mitochondrial biogenesis assay.

Cyprotex delivers consistent, high quality data with the flexibility to adapt protocols based on specific customer requirements.

• Mitochondrial proteins are encoded by both mitochondrial and nuclear genomes. Mitochondrial biogenesis is defined as the growth and division of pre-existing mitochondria¹. Certain drugs are able to affect mitochondrial biogenesis through inhibition of mtDNA replication or mitochondrial encoded protein synthesis.
• There is certain similarity between mitochondrial biogenesis and bacterial and viral replication. As a consequence, many antibacterial and antiviral agents cause mitochondrial toxicity through effects on human mitochondrial biogenesis. In fact, the FDA suggests that all antiviral drugs should be tested for effects on mitochondrial function².
• Cyprotex use high content screening and fluorescently labeled antibodies to evaluate ratios of an mtDNA-encoded protein (COX-1, a subunit of Complex IV) and an nDNA-encoded protein (SDH-A, a subunit of Complex II).
• The mitochondrial biogenesis assay can be used to identify drugs cause toxicity through inhibition of mitochondrial biogenesis.

Cyprotex’s mitochondrial biogenesis assay uses high content imaging to detect COX-1 (mtDNA-encoded) and SDH-A (nDNA-encoded) protein expression. Using the ratio of COX-1/SDH-A protein expression, specific effects on mitochondrial biogenesis can be identified. The positive control for the assay, chloramphenicol (antibiotic), inhibits mitochondrial biogenesis in a dose-dependent manner (figure 1).

_{1) Jornayvaz FR and Shulman GI (2010) Regulation of mitochondrial biogenesis. Essays Biochem 47; 69-84
2) Food and Drug Administration. Guidance for Industry: Antiviral product development - Conducting and submitting virology studies to the agency
3) Moreira AC et al, (2011) Mitochondria as a biosensor for drug-induced toxicity - Is it really relevant? Biosensors for Health, Environment and Biosecurity, Serra PA (Ed.); 411-444}