The hyperinflammatory state: increasing proclivity to develop necrotizing enterocolitis

Author(s):
Katherine Snyder; Alena Golubkova; Tyler Leiva; Heather Liebe; Camille Schlegel; Catherine Hunter

Background:

Necrotizing enterocolitis (NEC) is a devastating disease of premature neonates with significant morbidity and mortality. NEC is associated with prematurity, a hyperinflammatory response, and dysregulation of intestinal barrier function.

Hypothesis:

We hypothesize that NEC patients with a greater degree of prematurity will have an increased hyperinflammatory intestinal response compared to those without NEC.

Methods:

Enteroids were generated from intestinal tissue from varying degrees of premature neonates undergoing resection. They were treated in triplicate with 100ug/mL lipopolysaccharide (LPS), subjected to 24 hours of hypoxia inducing experimental NEC, then compared with untreated controls. Gene and protein expression of Tumor Necrosis Factor (TNF-α) and interleukin 8 (IL-8) were evaluated via RT-qPCR and ELISA to measure inflammatory response. ANOVA determined statistical significance (p<0.05).

Results:

Enteroids from NEC tissue that were treated with LPS + hypoxia expressed significantly higher levels of IL-8 (RTqPCR (p=0.003), ELISA (p=0.0002)) when compared to untreated NEC-derived enteroids with an increase in inflammatory marker concentration in those with a greater degree of prematurity (ELISA (p=0.0015)). A significantly higher level of IL-8 was seen in NEC-derived enteroids compared to control enteroids after treatment (RT-qPCR (p=0.024)) with a positive trend seen on ELISA. TNF-α levels were significantly elevated in NEC-derived enteroids following treatment compared to untreated NEC-derived enteroids (RTqPCR (p=0.006) and ELISA (p=0.002)) and compared to non-NEC-derived enteroids that underwent treatment on RTqPCR (p=0.025) and ELISA (p<0.0001).

Conclusions:

Enteroids generated from neonates with NEC have an elevated hyperinflammatory response in response to NEC-inducing stimuli compared with controls. Enteroids generated from neonates with NEC with a greater degree or prematurity have a larger increase in inflammatory markers. This tendency towards a hyperinflammatory state may be correlated with an infant’s proclivity to develop NEC and further demonstrates the hyperinflammatory state of prematurity. Further research could lead to strategies to identify and treat at-risk infants.