Early Protein Markers of Necrotizing Enterocolitis in Plasma of Preterm Pigs Exposed to Antibiotics

Abstract

Background: Most hospitalized preterm infants receive antibiotics in the first days of life to prevent or treat infections. Short-term, early antibiotic treatment may also prevent the microbiota-dependent gut inflammatory disorder, necrotizing enterocolitis (NEC). It remains a challenge to predict NEC, and a few early blood diagnostic markers exist. Using preterm pigs as model for infants, blood parameters and plasma proteins affected by early progression of NEC were profiled in preterm pigs subjected to oral, systemic, or no antibiotics after preterm birth. Methods: Preterm newborn pigs were treated with saline (CON) or antibiotics (ampicillin, gentamicin, and metronidazole) given enterally (ENT) or parenterally (PAR), and fed formula for 4 days to induce variable microbiome-dependent sensitivities to NEC. The gut was collected for macroscopic scoring of NEC lesions and blood for hematology, blood biochemistry, and LC/MS-based plasma proteomics. Statistical modeling was applied to detect plasma proteins affected by NEC and/or antibiotics. Results: Analyzed across different antibiotic regimens, NEC progression was associated with altered blood parameters and abundance of 89 plasma proteins that were functionally involved in extracellular membrane destruction, lipid metabolism, coagulopathy, and acute phase response. Large NEC-related changes were observed in abundance of RBP4, FGA, AHSG, C5, PTPRG, and A-1-antichymotrypsin 2, indicating potential serving as early markers of NEC. Conversely, antibiotic treatment, independent of NEC, affected only 4 proteins with main differences found between ENT and CON pigs. Conclusion: Early postnatal development of NEC lesions is associated with marked plasma protein changes that may be used for early NEC diagnosis.

Porcine Models of the Intestinal Microbiota: The Translational Key to Understanding How Gut Commensals Contribute to Gastrointestinal Disease

“…As gastrointestinal researchers continue to embrace the pig model and HMA pigs become increasingly accessible, emerging translational studies will divulge the pathogenesis of and putative therapies for diseases characterized by or associated with intestinal dysbiosis. Pig models have already proven advantageous in the study of microbiota-associated diseases such as necrotizing enterocolitis of infants, which is a complication of preterm, very-low birth weight infants that has been associated with bacterial colonization of the intestines (62)(63)(64)(65)(66)(67)(68)(69)(70). Particularly now that the porcine intestinal microbiota has been fully characterized through multiple life stages, gastrointestinal researchers can easily track changes in the microbial composition and make direct associations between these changes and disease (28,38,41,71).…”

The Role of the Tumor Suppressor Gene Protein Tyrosine Phosphatase Gamma in Cancer

“…Besides the classic whole structure of the phosphatase, the truncated form of the extracellular domain seems to be of particular relevance. Indeed, increasingly evidence emphasizes a role in several tissues under inflammatory state ( Lorenzetto et al, 2014 ; Moratti et al, 2015 ; Jiang et al, 2020 ). As previously discussed, PTPRG is regulated also in the context of myeloid cell differentiation ( Lissandrini et al, 2006 ).…”

Plasma Metabolomics to Evaluate Progression of Necrotising Enterocolitis in Preterm Pigs

“…The collected gastrointestinal tracts were divided into five regions (stomach, proximal, middle and distal small intestine, and colon) for macroscopic scoring of NEC lesions using our previously validated scoring system [ 19 ], where score 1 represents absence of macroscopic haemorrhage, oedema or mucosal abnormality; score 2 represents local hyperaemia; score 3 represents hyperaemia, extensive oedema and local haemorrhage; score 4 represents extensive haemorrhage; score 5 represents local necrosis and pneumatosis intestinalis; and score 6 represents extensive transmural necrosis and pneumatosis intestinalis. Representative pictures of the gut tissues can be seen in our previous publication [ 21 ]. The maximal NEC score across the five regions was used to classify the pigs into No-NEC (NEC score 1–2, n = 28), Mild-NEC (NEC score 3–4, n = 9) and Severe-NEC (NEC score 5–6, n = 10) groups for statistical analysis.…”

“…In this study a preterm pig model of NEC was used to investigate the plasma metabolites associated with NEC development and two different antibiotic protocols, oral or systemic antibiotic treatment for four days after birth. It has been previously reported in detail how such treatments, especially oral antibiotic treatment, reduced NEC sensitivity and improved gut structure/function, together with delayed gut bacterial colonisation and SCFA production [ 19 ], as well as the effects on systemic immunity [ 20 ] and plasma proteins [ 21 ]. We hypothesized that NEC development, independently or together with antibiotic treatment, would affect plasma metabolites.…”