Improved Characterization of 3D In Vitro Cell Culture Assays for the Drug Discovery Process

Project Summary The general drug discovery process can be described as a large funnel where thousands of compounds are screened for their efficacy and toxicological profile in the pursuit of developing a therapeutic with specific clinical propertiesThis funnel consists of a battery of biochemical and cellular assays for efficacy and toxicity of compoundsOne of the most important screening components of this funnel is the use of inexpensive and high throughput in vitro cell culture models to evaluate likelihood of efficacy or toxicity in vivoAccurate in vivo correlation is crucial for in vitro models since failure to eliminate unsuccessful candidate compounds in early screens results in failure in more expensive animal human trialsUntil recentlythe majority of in vitro drug screening was conducted on monolayer cell culturesD layers of cells are exposed to a compound of interest and evaluated through imagingflow cytometryor spectrophotometric assaysWhile this approach served as the basis for drug discovery for decadesthere are notable shortcomings in the ability of the results of these simple and inexpensive assays to translate in vivoThe complex three dimensional environment in vivo causes differences in response compared to monolayer cell cultures since monolayer models do not possess the diverse array of cellschemistriesand biochemical environments present in tissuesBecause of the well known limitations with traditional monolayer cell modelspharmaceutical companies have begun to rapidly adoptCAGRD in vitroD culture models since they have been shown to mimic the environment of in vivo tissues more accuratelyHoweverparadoxicallythe current techniques that companies are using to characterize theseD in vitro models do not fully utilize theD data that makes these models an improvement over traditionalD in vitro modelsEven the most comprehensive of high content confocal imaging is limited to visualizing only the surface layers of these modelsThe purpose of this project is to develop threeD in vitro cell culture assays that can be offered as a service to pharmaceutical companies for the drug discovery process that characterize the entirety of these modelsThese assays leverage the patented VisikolHISTO Mtissue clearing technique which has been shown to improveD cell culture characterization by allowing for afold increase in the amount of cells that can be detectedThese assays allow pharmaceutical companies to ask and answer complex mechanistic questions about the efficacy of their compounds which will accelerate the drug discovery process and get therapeutics to patients more quickly Project Narrative The purpose of this project is to develop threeD in vitro cell culture assays that can be offered as a service to pharmaceutical companies for the drug discovery processThese assays leverage the patented VisikolHISTO Mtissue clearing technique which has been shown to improveD cell culture characterization by allowing for afold increase in the amount of cells that can be detectedThese assays allow pharmaceutical companies to ask and answer complex mechanistic questions about the efficacy of their compounds which will accelerate the drug discovery process and get therapeutics to patients more quickly