A Versatile Human Intestinal Organoid-Derived Epithelial Monolayer Model for the Study of Enteric Pathogens.

Area of Science:

• Gastroenterology and Infectious Diseases
• Human Organoid Models
• Pathogen-Host Interactions

Background:

• Gastrointestinal infections are a major global health burden, with human-specific infection mechanisms poorly understood.
• Traditional models like animal studies have limitations in replicating the complexity of the human gut.
• Developing effective therapeutics and vaccines is hindered by the lack of physiologically relevant human models.

Purpose of the Study:

• To develop and validate a human intestinal organoid-derived epithelial monolayer (HIODEM) model for studying enteropathogens.
• To investigate pathogen tropism and infection patterns under in vivo-like conditions.
• To establish a platform for accelerating vaccine development against enteric pathogens.

Main Methods:

• Cultured two-dimensional epithelial monolayers from human intestinal organoids (terminal ileum, cecum, ascending colon).
• Utilized in vivo-like bacterial culturing conditions to induce pathogen virulence.
• Incorporated receptor activator of nuclear factor kappa-B ligand (RANKL) to enhance M cell differentiation.

Main Results:

• The HIODEM model successfully recapitulated intestinal epithelial composition, including enterocytes and goblet cells.
• RANKL treatment increased M cell presence, facilitating pathogen invasion.
• Distinct tissue tropism and infection patterns were observed for Shigella flexneri, Salmonella enterica serovars Typhi and Typhimurium, and pathogenic Escherichia coli.

Conclusions:

• The HIODEM model provides a powerful system for analyzing human-specific enteric infections.
• This model accurately reflects pathogen adherence, invasion, and tissue tropism.
• The system holds significant potential for future research into infection pathogenesis and therapeutic development.