1. Study of acute liver failure in children using next generation sequencing technology
- Author
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Hegarty, Rob and Thompson, Richard John
- Abstract
Acute liver failure (ALF) in children is a rare but potentially fatal condition without treatment. Diagnosis can be difficult and remains indeterminate in up to approximately half of cases. The study's aim was to identify undiagnosed, monogenic causes of ALF using targeted next generation sequencing (NGS). A capture library designed to target 64 genes known to cause ALF and/or a liver based metabolic disorder was constructed. Sequencing was carried out in 41 children who previously suffered from ALF without an identifiable aetiology. Biallelic variants were identified in 8 patients which may have been the cause of ALF or contributed to the clinical phenotype. Variants in NBAS and genes encoding mitochondrial proteins were the most common findings. Biallelic variants in NBAS is a cause of recurrent ALF triggered by fever. The pathobiology has been described to be due to disrupted retrograde transport between the Golgi and endoplasmic reticulum. However, it has also been suggested that disruption in nonsense mediated decay (NMD) may be involved. Skin fibroblasts from 4 children with biallelic variants in NBAS were used for the functional analyses including assessment of cell viability and protein expression. The gene expressions of GADD45A and GADD45B, endogenous NMD targets, were measured to assess whether NMD was disrupted. Cell viability was impaired in isolated cell culture conditions when placed under stress by glucose deprivation or incubation at 40°C. However, this was not consistent across all culture conditions. Western blot demonstrated reduced protein expression in 2 patients with two in-frame insertions. GADD45A and GADD45B gene expressions were not elevated in patient samples compared to controls suggesting that NMD is intact in these patients although disruption to a more localised NMD pathway remains a possibility. Next generation sequencing will be essential in improving diagnostic yield in future children with ALF. Using NBAS deficiency as an archetypal example of a monogenic cause of ALF insight was provided into its pathobiology. Progress is still required for the clinical application of NGS in children with ALF and future efforts should focus on overcoming the practical challenges.
- Published
- 2023