A Novel Model of P‐Glycoprotein Inhibitor Screening Using Human Small Intestinal Organoids

Abstract

P-glycoprotein (P-gp), an important efflux transporter in intestine, regulates the bioavailability of orally taken drugs. To develop an in vitro model that preferably mimics the physiological microenvironment of human intestine, we employed the three-dimensionally (3D) cultured organoids from human normal small intestinal epithelium. It was observed that the intestinal crypts could efficiently form cystic organoid structure with the extension of culture time. Furthermore, the physiological expression of ABCB1 was detected at both mRNA and protein levels in cultured organoids. Rhodamine 123 (Rh123), a typical substrate of P-gp, was actively transported across 3D organoids and accumulated in the luminal space. This transport process was also inhibited by verapamil and mitotane. In summary, the above-mentioned model based on human small intestinal 3D organoids is suitable to imitate the small intestinal epithelium and could be used as a novel in vitro model especially for P-gp inhibitor screening.

Organoid-derived Duodenum Intestine-Chip for preclinical drug assessment in a human relevant system

“…This is not feasible in pre-clinical species such as rat and dog due to marked species differences in the expression and regulation of cytochromes P450 as well as substrate specificity of the nuclear receptors, such as pregnane X receptor (PXR), responsible for the transcriptional regulation of CYP3A4 and several drug transporters. Caco-2 Intestine-Chip has been previously reported to possess an increased activity of the CYP450 enzymes when compared to the conventional static culture of Caco-2 cells on transwell insert (25). However, the gene expression level of CYP3A4 measured in this system is significantly lower than in the adult human intestine ( Figure 5A), limiting its application for pharmaceutical research, specifically pharmacokinetic evaluation.…”

“…Using these methods, organoids derived from all regions of the intestinal tract can be established 21,23 and applied into different areas of research including organ development, disease modeling, and regenerative medicine 24 . However, the characterization of the pharmacokinetic properties of this system, as well as the its validation for the use in drug discovery and development, is still very limited [25][26][27][28][29] . One potential limitation could be due to the substantial technical challenges associated with the use of this technology for ADME applications.…”

Improvement of paracellular transport in the Caco-2 drug screening model using protein-engineered substrates

“…Typically, these rely on addition of chemicals [16–20] or co-culture with other cell types [12,21–24]. Various three-dimensional (3D) drug screening models have also been developed, including organoid structures derived from primary tissue [25,26] and micro-fluidic, organ-on-a-chip devices [27]. While scientifically interesting, these strategies are technically cumbersome and not readily translatable to high-throughput settings.…”

Circulating tumor cell-derived organoids: Current challenges and promises in medical research and precision medicine

“…Organoids could be useful for testing drug efficacy, drug toxicity studies in liver organoids, or drug bioavailability studies in intestinal organoids [102,103]. In particular, CTC-derived models with relevant pathologies of patients could be a key link to screening specific drug/s [75,89,104].…”