Association between the p.Thr1406Asn polymorphism of the carbamoyl-phosphate synthetase 1 gene and necrotizing enterocolitis: A prospective multicenter study

Abstract

The p.Thr1406Asn (rs1047891) polymorphism of the carbamoyl-phosphate synthetase 1 (CPS1) gene has been linked to functional consequences affecting the downstream availability of the nitric oxide precursor L-arginine. L-arginine concentrations are decreased in preterm infants with necrotizing enterocolitis (NEC). In this multicenter prospective study, we investigated the association of the p.Thr1406Asn polymorphism with NEC in 477 preterm infants (36 cases of NEC) from 4 European neonatal intensive care units (Maastricht, Las Palmas de Gran Canaria, Mantova, and Milan). Allele and genotype frequencies of the p.Thr1406Asn polymorphism did not significantly differ between the infants with and without NEC. In contrast, the minor A-allele was significantly less frequent in the group of 64 infants with the combined outcome NEC or death before 34 weeks of corrected gestational age than in the infants without the outcome (0.20 vs. 0.31, P = 0.03). In addition, a significant negative association of the A-allele with the combined outcome NEC or death was found using the dominant (adjusted odds ratio, aOR: 0.54, 95% CI 0.29–0.99) and the additive (aOR 0.58, 95% CI 0.36–0.93) genetic models. In conclusion, our study provides further evidence that a functional variant of the CPS1 gene may contribute to NEC susceptibility.

Risk of Necrotizing Enterocolitis Associated With the Single Nucleotide Polymorphisms VEGF C-2578A, IL-18 C-607A, and IL-4 Receptor α-Chain A-1902G: A Validation Study in a Prospective Multicenter Cohort

“…Methods for statistical analysis were identical to those used in our previous study (16) and are therefore literally reproduced here. Categorical variables were expressed as counts or percentages and compared using the chi-square test.…”

“…The present study was performed on DNA samples collected during a previous study on carbamoyl phosphate synthetase SNPs as risk factor for NEC (16). The protocol of the above study was reviewed and approved by the institutional review board (IRB) for each participating center (see names below) and we obtained written informed consent from the parents.…”

“…Definitions were identical to those used in our previous study (16) and are therefore literally reproduced here. Data on clinical characteristics and outcomes were obtained from the medical records.…”

“…In the present study we aimed to confirm the association with NEC of three candidate SNPs: the VEGF C-2578A polymorphism (rs699947) (12,13), the IL-18 C-607A polymorphism (rs1946518) (14), and the IL-4 receptor αchain (IL-4Rα) A-1902G polymorphism (rs1801275) (15). We performed our investigation in a cohort of preterm infants from four neonatal intensive care units located in three different European countries (Spain, Italy, and the Netherlands) (16,17).…”

Carbamyol phosphate synthetase 1 gene (4217C>A) polymorphism and its relation to low plasma arginine level among preterm with necrotizing enterocolitis; a single center Egyptian study

“…The purified genomic DNA was stored at−80°C until use. The polymerase chain reaction were used to amplify a 214bp fragment encompassing the C-to-A nucleotide trans version at position 4217 of exon 36 of the gene-encoding CPS 1 using forward primer 5'-TAAATGCAGCTGTTTGCCAC-3' and reverse primer 5'GACTTGCAATCAAGTAAGGTGAAA-3' as designed by Moonen et al, 17 Polymerase chain reaction (PCR) amplification were carried out in a total volume of 25uL, containing 80ng genomic DNA, 12.5ul of 2 x Dream TaqTM Green Master Mix [Life Technologies, Eggenete in, Germany]and 25Pmol of each primer with cycling parameters as follows: initial denaturation at 95°C for 5min, 35 cycles at 94°C for 30s, annealing at 61°C for 30s and 72°C for 60s and one final cycle of extension at 72°C for 7min. Genotyping of the DNA sequence variants was performed using PCR-based RFLP assays through digestion of amplified PCR product of using specific restriction endonuclease enzymes according to manufacturer`s recommendation [New England Bio labs, Sch Wallach, Germany].…”