The Risk of Developing Liver Fibrosis is Associated with Polymorphism and Rare Haplotypes of the TGFB1 Gene in Children with End Stage Liver Disease

Aim: to determine the distribution of the three most significant single nucleotide polymorphisms (SNPs) of the TGFB1 gene (rs1800469, rs1800470, rs1800471) and their haplotypes in children with liver fibrosis.

Materials and methods. The study included 107 pediatric liver recipients (45 boys, 62 girls) aged from 3 to 73 months (median — 8 months). The control group consisted of 199 healthy individuals (78 men, 121 women) aged 32.7 ± 9.6 years. During histological examination of the liver removed before transplantation, fibrosis of different severity grades was diagnosed in all children in accordance with the criteria of the METAVIR scale: 5 cases — Grade F1, 9 cases — Grade F2, 14 cases — Grade F3 and 79 cases — Grade F4. The indication for liver transplantation was end-stage liver disease: biliary atresia (n = 61) and hypoplasia (n = 8), Alagille syndrome (n = 8), Caroli disease (n = 8), Byler disease (n = 6) and other liver diseases (n = 16). SNPs were determined by real-time polymerase chain reaction using TaqMan probes in genomic DNA, isolated from peripheral blood.

Results. In children with liver fibrosis of different severity grades, the frequency distribution of the studied TGFB1 gene SNPs was: for rs1800469 — 38 % GG homozygotes, 42 % AG heterozygotes and 20 % AA homozygotes; for rs1800470 — 50 % AA, 29 % AG, 21 % GG; for rs1800471 — 93 % CC, 7 % GC, 0 % GG. The distribution of SNPs rs1800469 and rs1800471 corresponded to the Hardy — Weinberg equilibrium and did not differ from that in healthy individuals. The distribution of rs1800470 in children with fibrosis, in contrast to healthy controls, did not correspond to the Hardy — Weinberg law (p = 0.00026). For the studied SNPs, linkage disequilibrium was shown; five main combinations were observed: three haplotypes, including two most common ones, were distributed, in total, in about 55 % of children with fibrosis and 91 % of healthy individuals; these frequencies were not statistically different in the group of patients and healthy individuals. Significant differences were detected in the distribution of two rarer haplotypes — A-A-C and G-G-C (respectively rs1800469, rs1800470, rs1800471), which were observed more often in patients with liver fibrosis than in healthy individuals: respectively, in 6.03 (95% CI: 3.06–11.89; p < 0.0001) and 3.71 (95% CI: 1.94–7.08; p = 0.0001) times.

Conclusions. In children with liver fibrosis, the distribution of single nucleotide polymorphism rs1800470 and two rare haplotypes rs1800469, rs1800470, rs1800471 of the TGFB1 gene differs significantly from that in healthy individuals. Polymorphism of rs1800470, as well as haplotypes A-A-C or G-G-C at positions rs1800469, rs1800470, rs1800471, may predispose to the development of liver fibrosis in children with liver failure.

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