Molecular and Functional Characterization of Human Intestinal Organoids and Monolayers for Modeling Epithelial...

Area of Science:

• Gastroenterology and Hepatology
• Cell Biology
• Drug Discovery

Background:

• Patient-derived organoids (PDOs) show promise for inflammatory bowel disease (IBD) research but face challenges in consistent differentiation and functional characterization.
• Standardized molecular and cellular profiling is needed to validate PDO models for IBD drug discovery.

Purpose of the Study:

• To profile molecular and cellular features of intestinal organoid models.
• To examine the differentiation and functional barrier establishment of patient-derived organoids (PDOs) and monolayers.
• To assess the potential of PDOs for developing targeted therapies for IBD.

Main Methods:

• Generated patient-derived organoids (PDOs) and monolayers from control and IBD patients.
• Performed molecular and functional profiling, including transcriptional response analysis.
• Assessed epithelial barrier function using transepithelial electrical resistance (TEER) and response to inflammatory cytokines (IFN-γ, TNF-α).
• Evaluated the efficacy of tofacitinib in preventing cytokine-induced barrier disruption.

Main Results:

• Organoids and monolayers differentiated into mature intestinal epithelial cell types, forming functional barriers with sustained TEER.
• Inflammatory cytokines IFN-γ and TNF-α compromised barrier integrity.
• Tofacitinib demonstrated dose-dependent inhibition of cytokine-induced barrier damage.

Conclusions:

• Developed and characterized human colonic and ileal organoid models capable of functional differentiation.
• Demonstrated that PDOs form robust epithelial barriers in monolayer culture, responding to pharmacological modulation.
• Confirmed consistent transcriptional and functional profiles in control and IBD patient-derived organoids, supporting their use in developing epithelial-targeted therapies for IBD.