High level of oxysterols in neonatal cholestasis: a pitfall in analysis of biochemical markers for Niemann-Pick type C disease

Giulia Polo 1 , Alessandro Burlina 2 , Francesca Furlan 1 , Thilini Kolamunnage 1 , Mara Cananzi 3 , Laura Giordano 1 , Martina Zaninotto 4 , Mario Plebanihttp://orcid.org/0000-0002-0270-1711 4 , and Alberto Burlina 1
  • 1 Division of Inherited Metabolic Diseases, Department of Women and Children’s Health, University Hospital of Padua, Padua, Italy
  • 2 Neurology Unit, St. Bassiano Hospital, Bassano del Grappa, Italy
  • 3 Paediatric Gastroenterology Unit, Department of Women and Children’s Health, University Hospital of Padua, Padua, Italy
  • 4 Department Laboratory Medicine, University Hospital of Padua, Padua, Italy
Giulia Polo, Alessandro Burlina, Francesca Furlan, Thilini Kolamunnage, Mara Cananzi, Laura Giordano, Martina Zaninotto, Mario PlebaniORCID iD: http://orcid.org/0000-0002-0270-1711 and Alberto Burlina

Abstract

Background: Niemann-Pick disease type C (NPC) is a rare lipid storage disorder characterized by progressive neurological deterioration. Diagnosing NPC is challenging as clinical signs and symptoms are variable and non-specific. Two oxysterols, cholestane-3β,5α,6β-triol (triol) and 7-ketocholesterol (7KC), have been proposed as biomarkers for aiding diagnosis of NPC. This study evaluated the use of triol and 7KC as biomarkers in cholestatic neonates with suspected NPC.

Methods: Plasma triol and 7KC were analysed as dimethylglycine esters using an liquid chromatography – tandem mass spectrometry (LC-MS/MS) assay in selected neonates with severe cholestasis and suspected NPC (n=7), adults with cholestasis (n=15), patients with confirmed NPC (positive controls; n=11 [one child and 10 adults]), healthy subjects (negative controls; n=40 [20 children and 20 adults]), and cholestatic adults (comparative reference; n=15). The LC-MS/MS method was subjected to a number of tests for accuracy and consistency.

Results: Triol and 7KC levels were substantially and significantly increased in NPC positive patients compared with healthy controls (p<0.001). However, positive results (markedly increased levels of both oxysterols) were identified in 6/7 (86%) neonates with cholestasis. Genetic testing confirmed NPC only in one neonate who had increased triol and 7KC, and increased oxysterol levels among neonates with no identified NPC gene mutations were considered likely due to biliary atresia (BA).

Conclusions: While the potential of oxysterols as NPC biomarkers has been well evaluated in older patient populations (without cholestasis), our data suggest that cholestasis might represent a pitfall in oxysterol measurements intended to aid diagnosis of NPC in affected patients.

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