Age disparities in intestinal stem cell quantities: a possible explanation for preterm infant susceptibility to necrotizing enterocolitis

Abstract

Purpose: Preterm infants are more susceptible to necrotizing enterocolitis (NEC) than term Queryinfants. This may be due to a relative paucity of Lgr5⁺ or Bmi1⁺-expressing intestinal stem cells (ISCs) which are responsible for promoting intestinal recovery after injury. We hypothesized that the cellular markers of Lgr5⁺ and Bmi1⁺, which represent the two distinct ISC populations, would be lower in younger mice compared to older mice. In addition, we hypothesized that experimental NEC would result in a greater loss of Lgr5⁺ expression compared to Bmi1⁺ expression.

Methods: Transgenic mice with EGFP-labeled Lgr5 underwent euthanasia at 10 different time points from E15 to P56 (n = 8-11/group). Lgr5⁺-expressing ISCs were quantified by GFP ELISA and Bmi1⁺ was assessed by qPCR. In addition, Lgr5^EGFP mice underwent experimental NEC via formula feeding and hypoxic and hypothermic stress. Additional portions of the intestine underwent immunostaining with anti-GFP or anti-Bmi1⁺ antibodies to confirm ELISA and PCR results. For statistical analysis, p < 0.05 was significant.

Results: Lgr5⁺ and Bmi1⁺expression was lowest in embryonal and early postnatal mice and increased with age in all segments of the intestine. Experimental NEC was associated with loss of Lgr5⁺-expressing ISCs but no significant change in Bmi1⁺ expression.

Conclusion: Lgr5⁺ and Bmi1⁺ expression increase with age. Lgr5⁺-expressing ISCs are lower following experimental necrotizing enterocolitis while Bmi1⁺ expression remains relatively unchanged. Developing a targeted medical therapy to protect the low population of ISCs in preterm infants may promote tissue recovery and regeneration after injury from NEC.

Keywords: Animal study; Bmi1; Intestinal stem cells; Lgr5; Necrotizing enterocolitis.