| Abstract
| - Introduction. Serum protein N-glycosylation has previously been shown to distinguish adult patients with simple steatosis from those with non-alcoholic steatohepatitis (NASH). The pattern of the disease in paediatric patients is distinct from adults. Aim. The aim of this study was to characterise the glycomic profile of children with varying degrees of NAFLD to identify potential biomarker profiles of disease. Method. Children with biopsy proven non-alcoholic fatty liver disease (n=51) were recruited from a tertiary paediatric hepatology unit. Liver biopsy was scored according to NAFLD activity score. Blood was taken on day of biopsy for analysis. Serum protein N-glycosylation patterns were assessed with DNA-sequencer assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) and compared with histology. Results. Median age at biopsy was 13.3 years (range 4.5-17.4). 31 were male. Median BMI z-score was 1.81. 23 children scored as simple steatosis/borderline NASH and 28 as true NASH. 18 children had no/minimal fibrosis (< F2) and 33 had significant fibrosis (≥ F2). Similar to previous work in adult patients with NAFLD, peak 1 (NGA2F) was the most significantly raised N-glycan in paediatric NASH patients with peak 5 (NA2) demonstrating the greatest decrease. The logarithmically transformed ratio of peak 1 to peak 5 (Glycomarker) was −0.85 (SD 0.22) in simple steatosis/borderline NASH and −0.73 (SD 0.12) in NASH (p=0.02). The biomarker correlated well with the amount of lobular inflammation with a consistent increase with ascending grade of inflammation. There was also a trend towards significance in differentiating patients with significant fibrosis ≥ F2; −0.74 (SD 0.13) from patients with no/minimal fibrosis < F2; −0.86 (SD 0.24), (p=0.06). Glyco-analysis of immunoglobulin G (IgG) confirmed the undergalactosylation status with a significant increase in peak 1 (NGA2F; p=0.024) and a significant decrease of peak 6 (NA2F; p=0.01) on IgG. In multivariate analysis of the Glycomarker, GGT, AST and INR, only the Glycomarker displayed a significant result for distinguishing simple steatosis from NASH (p=0.019). Conclusion. In conclusion, the findings in this study are novel in that they represent the first Glycomic analysis of paediatric NAFLD. They validate findings in adults in that a Glycomarker can serve reliably as a biomarker of severity of disease in NAFLD. The same N-glycosylation alterations are observed in paediatric NASH patients when compared to an adult population and therefore the same biomarker can be used. B cells play a dominant role in the N-glycan alterations of NASH patients, both in an adult and paediatric population.
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