High-throughput IBD drug screening methods are crucial for identifying new therapeutics. Tumor Necrosis Factor Alpha (TNF-a) is a potent cytokine. It is over-expressed by cells during chronic intestinal inflammation. Furthermore, TNF-a represents the most validated clinical target for ulcerative colitis (UC) and inflammatory bowel diseases (IBD). Several emerging therapeutics target this cytokine for inactivation to reduce inflammation. TNF-a activation of intestinal epithelia can disrupt the intestinal barrier through various mechanisms. This includes the activation of inflammatory master regulators like NFkB-induced IL-8 production and secretion. It also has direct effects on tight junction integrity and cellular apoptosis. Therefore, this illustrates the importance of high-throughput IBD drug screening methods.
A Novel, High-Throughput Platform to Characterize Novel IBD Therapeutics
Altis’ RepliGut® Planar model provides a novel, human primary culture cell system. This system reprises the cellular complexity of the intestinal tract in vivo. It is established using 12- and 96-well Tranwell® inserts. These inserts maximize throughput and dosing/assay flexibility, making it well-suited for high-throughput IBD drug screening.
Altis has established a model of the inflammatory response to TNF-a using RepliGut® planar. It is designed to enable the screening and detailed characterization of novel therapeutic agents. This is done by combining measures that bridge non-invasive and high-throughput testing with clinically relevant endpoints of the disease. This innovative approach forms the backbone of high-throughput IBD drug screening processes.
In this model, TNF-a produces a dose-dependent decrease in TEER (electrical resistance). Concurrently, it increases the permeability of organic molecules across the epithelium, which are highly correlated. This establishes a predictable response between TEER and the luminal permeability of surrogates of bacterial toxins.
Furthermore, the planar format also enables easy sampling of the basolateral cell media. This media can be used to assay epithelial production of cytokines, including IL-8 (the major neutrophil-chemoattractant produced downstream of TNF-a). Collectively, the RepliGut® planar model enables testing of novel IBD therapeutics. Tests can be run in a 96-well plate format. This format can include relevant measures of barrier function and inflammation. Consequently, high-throughput IBD drug screening is significantly enhanced.
Finally, as the experimental platform is highly flexible, additional endpoints including immunofluorescence, gene expression, and cell viability can be easily incorporated.