Jejunum-derived NF-κB reporter organoids as 3D models for the study of TNF-alpha-induced inflammation

Inflammation is an important process for epithelial barrier protection but when uncontrolled, it can also lead to tissue damage. The nuclear factor-kappa light chain enhancer of activated B cells (NF-kappa B) signaling pathway is particularly relevant in the intestine, as it seems to play a dual role. Whereas NF-kappa B protects intestinal epithelium against various noxious stimuli, the same pathway mediates intestinal inflammatory diseases by inducing pro-inflammatory gene expression. The availability of appropriate in vitro models of the intestinal epithelium is crucial for further understanding the contribution of NF-kappa B in physiological and pathological processes and advancing in the development of drugs and therapies against gut diseases. Here we established, characterized, and validated three-dimensional cultures of intestinal organoids obtained from biopsies of NF-kappa B-RE-Luc mice. The NF-kappa B-RE-Luc intestinal organoids derived from different intestine regions recreated the cellular composition of the tissue and showed a reporter responsiveness similar to the in vivo murine model. When stimulated with TNF-alpha, jejunum-derived NF-kappa B-RE-Luc-reporter organoids, provided a useful model to evaluate the anti-inflammatory effects of natural and synthetic compounds. These reporter organoids are valuable tools to explore the epithelial TNF-alpha-induced NF-kappa B contribution in the small intestine, being a reliable alternative method while helping to reduce the use of laboratory animals for experimentation.

Automated summary

The NF-kappa B signaling pathway plays a crucial role in the intestine, protecting epithelial cells from harm while also mediating inflammatory diseases. Established NF-kappa B reporter organoids provide a valuable tool for evaluating anti-inflammatory compounds and exploring their effects in the small intestine.