Optimising the induction of inflammation within preterm infant-derived intestinal epithelial organoids

Area of Science:

• Gastroenterology
• Microbiology
• Developmental Biology

Background:

• Preterm infants (<32 weeks gestation) exhibit gut microbial dysbiosis and inflammation.
• Preterm infant-derived intestinal organoids (PIOs) offer a promising model for studying gut microbiome-host interactions.
• Understanding inflammatory responses in the preterm gut is crucial for improving infant health outcomes.

Purpose of the Study:

• To optimize an in vitro inflammation model using PIOs.
• To investigate host-microbe interactions and inflammatory pathways in the preterm gut.
• To establish a framework for studying gut inflammation in preterm infants.

Main Methods:

• Developed an anaerobic co-culture system for PIO monolayers, mimicking intestinal oxygen gradients.
• Tested various inflammatory stimuli, including pathobionts, lipopolysaccharide (LPS), and flagellin.
• Optimized stimulus application (apical LPS, basolateral flagellin) and incubation time (3 hours) for maximal response.

Main Results:

• A combination of apical LPS and basolateral flagellin induced a robust inflammatory response in PIOs.
• Key indicators included enhanced pro-inflammatory cytokine secretion and chemokine-driven immune cell recruitment.
• Observed activation of TNFα and IL17C pathways, a shift from NF-κB to AP-1 signaling, and tissue remodeling.

Conclusions:

• The optimized PIO inflammation model provides a reliable platform for studying preterm infant gut inflammation.
• This model facilitates research into the complex interplay between the microbiome and host in health and disease.
• The findings offer a framework for future studies on preterm infant gut pathophysiology and therapeutic interventions.