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Licensed Unlicensed Requires Authentication Published by De Gruyter December 9, 2015

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

  • Giulia Polo , Alessandro Burlina , Francesca Furlan , Thilini Kolamunnage , Mara Cananzi , Laura Giordano , Martina Zaninotto , Mario Plebani ORCID logo and Alberto Burlina EMAIL logo

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.


Corresponding author: Dr. Alberto Burlina, MD, Division of Inherited Metabolic Diseases, Department of Women and Children’s Health, University Hospital of Padua, Italy, E-mail:

Acknowledgments

We thank Dr. Lucia Santoro (Department of Pediatrics, University Hospital Ancona, Italy), Dr. Marina Scarlato (Department of Neurology, San Raffaele Hospital, Milan, Italy) and Dr. Olimpia Musumeci (Department of Neuroscience, University of Messina, Italy) for providing plasma samples from NPC patients. Matthew Reilly PhD provided medical writing assistance in the preparation of this manuscript for submission. This work was supported by the Cometa-A.S.M.M.E (Associazione per lo Studio delle Malattie Metaboliche Ereditarie, Padova Italy) and Actelion Pharmaceuticals Ltd.

Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

Research funding: GP, APB and ABB have received travel expenses from Actelion Pharmaceuticals Ltd. FF, TK, MC, LG, MZ and MP have no conflicts to declare.

Employment or leadership: None declared.

Honorarium: None declared.

Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Supplemental Material

The online version of this article (DOI: 10.1515/cclm-2015-0669) offers supplementary material, available to authorized users.


Received: 2015-7-14
Accepted: 2015-10-31
Published Online: 2015-12-9
Published in Print: 2016-7-1

©2016 by De Gruyter

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