Persistent Post Recovery Hyperinflammation of NEC is Ameliorated by 5-ASA Treatment

Author(s):
Katherine Snyder; Chase Calkins; Cody Dalton; Grant Gershner; Camille Schlegel; Catherine Hunter

Background:

Necrotizing enterocolitis (NEC) is the leading gastrointestinal cause of death of premature neonates. We have previously shown that this hyperinflammatory state persists even post-recovery.

Hypothesis:

We hypothesize that recovered NEC patients will have a decreased hyperinflammatory response when the anti-inflammatory medication mesalamine (5-ASA) is administered even when exposed to in-vitro NEC induction.

Methods:

Intestinal specimens were collected from patients that have recovered from NEC and generated into enteroids. Enteroids were treated with lipopolysaccharide (LPS), subjected to 24 hours of hypoxia, then compared with untreated controls. In each arm, one half were subjected to 5-ASA treatment during the experimental timeframe. Expression of Tumor Necrosis Factor (TNF-α) and interleukin 8 (IL-8) were evaluated via RTqPCR. ANOVA determined statistical significance (p<0.05).

Results:

Following NEC inducing in-vitro treatment, recovered NEC enteroids demonstrate elevated levels of inflammatory markers compared to recovered NEC enteroids that did not undergo treatment (p=0.002 TNF-α, up-trend IL-8). Recovered NEC enteroids that were treated with 5-ASA and subjected to NEC induction show a significant decrease in inflammatory markers compared to those that were not given 5-ASA (p=0.0014 TNF-α, down-trend IL-8). There is no significant difference between recovered NEC enteroids exposed to 5-ASA and NEC induction compared to recovered NEC enteroids without NEC induction (p=0.7383 TNF-α, p=0.3913 IL-8).

Conclusions:

Following NEC induction, there is a significant upregulation of hyperinflammatory genes in enteroids from patients that have recovered from NEC. Recovered NEC enteroids that are exposed to 5-ASA during NEC induction have no significant difference in the inflammatory marker levels when compared to enteroids that did not undergo induction. This decrease in a hyperinflammatory response despite NEC induction may demonstrate the impact that anti-inflammatory agents could have following NEC recovery. This could be important given the previously demonstrated robust hyperinflammatory response to a second hit after recovery and may impact the trajectory of an illness post-recovery from NEC.