PROCOMCURE Biotech’s molecular toxicology (MOLTOX)
…gene expression as a screening tool for the evaluation of a compound’s potential toxicity.
For many candidate molecules extracted from cell-based assays with highly variable drug potential, there is a double-edged, intrinsically inseparable transition from biological activity to cytotoxicity. The early monitoring or rather exclusion of a lead compound’s (hepato-) toxicity represents a crucial step in the drug discovery and development pipeline for saving time, costs, and, most importantly, reducing the likelihood of late stage failures. The relatively new field of ‚toxicogenomics‘ integrates state-of-the-art transcriptional profiling into conventional toxicological science, and so constitutes an attractive approach both to predict toxicity (long before functional damage is observed) and gain a mechanistic understanding of underlying molecular alterations . At this juncture, PROCOMCURE’s proprietary MOLTOX platform built upon respective achievements applies real-time quantitative reverse transcription PCR for large-scale, sensitive quantification of mRNA transcripts differentialy expresssed in response to (toxic) agent / compound exposure.
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- PROCOMCURE Biotech’s comprehensive MOLTOX real-time PCR assay panel reflects a literature-deduced compilation of approximately 400 human (independent or interrelated) key genes – amongst them phase I and II drug metabolizing enzymes; chaperones; heat shock proteins, stress response genes; drug transporters; etc. – participating in 14 different ADME-relevant pathways possibly activated in response to drug exposure. Amongst analyzed cascades are: apoptosis / necrosis, fatty acid metabolism, oxidative stress / antioxidant response, mitochondrial energy metabolism, DNA damage / repair, immunotoxicity, hepatoxicity, steatosis, cholestasis, and cytochrome P450s. An entire list of genes (and their categorization) can be downloaded here.
- Cells are cultivated until they reach 80% confluence, exposed to respective agents under investigation in relation to vehicle (mostly DMSO) control for 24 h, and the prepared total RNA finally converted into cDNA using established procedures.
- Differential gene expression changes associated with pathway activation or inhibition are measured from resultant templates by optimized RTq-PCR in high-throughput compatible 384-well format on an Applied Biosystems® ViiA™ 7 real-time system, taking advantage of PCC‘s fast turnaround screening services.
- PROCOMCURE Biotech’s MOLTOX provides a predictive indicator of potential drug toxicity, and, so, delivers more informative data than traditional cellular / biochemical assays alone.
- Utmost detection sensitivity permits to assay wide dynamic range responses (pM to µM concentrations). Experiments are performed in biological triplicates each, and statistical significance / fold inductions assessed via StatMiner® (Integromics®).
- Once the exploratory work is complete, more focused experiments can be carried out.
Cell lines offered by PROCOMCURE Biotech
|Cell type||Cell line||Tissue|
|epithelial||AD HEK 293||kidney|
|fibroblasts||> 70 primary cell lines||skin|
|endothelial||HMEC-1||dermal microvascular endothelium|
|endothelial||HUVEC||umbilical vasc. endothelium|
|gingiva||> 50 primary cell lines||dental|
|pulpa||> 50 primary cell lines||dental|
|cyste||> 50 primary cell lines||dental|
|stem cells||> 60 primary cell lines||subcutaneous/visceral fat|
 Blomme EA et al. 2009: Use of toxicogenomics to understand mechanisms of drug-induced hepatotoxicity during drug discovery and development; Gatzidou E et al. 2007: Toxicogenomics: a pivotal piece in the puzzle of toxicological research; Waters M et al. 2004: toxicogenomics and systems toxicology: aims and prospects; Nuwaysir EF et al. 1999: microarrays and toxicology: the advent of toxicogenomics