A novel live biologic for the prevention and treatment of NEC-induced neurodevelopmental impairment

SUMMARY: Worldwide, up to 25% of premature infants born prior to 31 weeks gestation, and up to 50% of those born prior to 28 weeks gestation, have neurodevelopmental impairment (NI). The causes of NI are multifactorial, but neuroinflammation is now recognized as a key component. Early life stresses commonly encountered by premature infants, including hypoxia, forced formula feeding, and hypothermia, have the potential to trigger inflammation throughout the body. This inflammation can originate in the intestine, e.g. during neonatal necrotizing enterocolitis (NEC), where the microbiota incite a dysregulated inflammatory response. Excessive inflammation in the periphery propagates neuroinflammation and subsequent long-term NI. We have developed a novel, tunable probiotic delivery system in which the probiotic Limosilactobacillus reuteri (Lr) is administered in a protective biofilm on biocompatible porous microspheres that provide Lr with cargos that induce beneficial properties. Administration of Lr in this biofilm state reduces the incidence of NEC and subsequent neuroinflammation in a rat model after just a single dose. The central hypothesis of this proposal is that identifying the extent to which Lr antimicrobial and/or anti-inflammatory properties attenuates NI can be leveraged to tune these Lr properties to optimally prevent, or even treat, NEC-induced NI. Studying the neurodevelopmental impacts of optimized dosing regimens on both NEC and healthy control pups will demonstrate the contributions of these properties towards not only preventing and treating NEC-induced NI, but also healthy neurodevelopment. These studies are in line with the recent NOFO (PAR-23-130) for Translational Research in Maternal and Pediatric Pharmacology and Therapeutics, where we intend to develop an FDA approved probiotic formulation which can be administered to all premature infants at risk of developing NEC. AIM 1: Determine the extent to which Lr antimicrobial pathways can prevent NEC-induced neurodevelopmental impairment (NI). Bacterial dysbiosis can incite inflammatory responses in the periphery and brain. We hypothesize that tuning Lr antimicrobial activity can prevent NEC-induced NI. AIM 2: Determine the extent to which Lr anti-inflammatory pathways can prevent NEC-induced NI. We will determine the extent to which Lr anti-inflammatory pathways prevent neuroinflammation and subsequent NI. We hypothesize that tuning Lr anti-inflammatory properties can prevent NEC-induced NI. AIM 3: Determine the dominant Lr-dependent pathways for the prevention and treatment of NI using all proven cargos. We will determine whether Lr antimicrobial or anti-inflammatory pathways are dominant, antagonistic, additive or synergistic in the prevention and treatment of NI. We hypothesize that tuning the best combinations of Lr pathways can be used to both prevent and treat NEC-induced NI. The significance of the proposed research is that it will lead to a better understanding of the ability of Lr to prevent and treat NEC-induced NI, leading to improved novel therapeutic approaches.