NORTHSHORE UNIVERSITY HEALTHSYSTEM RESEARCH INSTITUTE

Necrotizing enterocolitis is a common and devastating intestinal emergency that primarily affects premature infants, whose etiology, pathogenesis, and treatment strategies remain poorly understood. Findings in our lab and others has identified an important role for PAF (platelet activating factor), an endogenous phospholipid pro-inflammatory mediator, in the pathogenesis of experimental NEC. The research was designed (1) To investigate the importance of TLR's in intestinal epithelium during ontogeny in NEC pathogenesis, utilizing TLR mutant and knockout mice and subjecting them to our neonatal mouse model of NEC, (2) To ivestigate mechanisms of PAF-induced TLR expression in enterocyts in vitro, focusing on the role of STAT3, NFkB, and inhibitory IkBa using cell culture transfection and siRNA techniques, and (3) To evaluate the influence of PAF on intestinal TLR in vivo using PAF-AH knockout and PAF receptor knockout mice. In the ARRA supplement, we are pursuing some exciting findings that were identified in the last year of the R01, namely that PAF-AH knockout animals are at higher risk for NEC but lower risk for early neonatal sepsis. The hypothesis is that mucosal PAF protects from bacterial translocation into the systemic circulation during the acquisition of normal bacterial colonization, and that the knockout animals have higher local PAF levels. To further evaluate these findings, we are making several double knockout animals, using TLR/PAF-AH knockouts and PAF-AH/PAFR knockouts. During the first quarter of this award, we have hired a laboratory associate and these double knockouts are in process.

Clarification of Codes