MSACL 2023 Abstract

Plasma and Platelet Lipidome Changes in Treated Fabry Disease Patients

Bo Burla (1) Jeongah Oh (2) Sock Hwee Tan (3,4) Nathalie Piraud (5) Mark Y. Chan (3,4) Anne K. Bendt (1) Federico Torta (2) Albina Nowak (6) Markus R. Wenk (1,2) Pierre–Alexandre Krayenbühl (6)
(1) Singapore Lipidomics Incubator, Life Sciences Institute, National University of Singapore, Singapore (2) Precision Medicine Translational Research Program and Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore (3) Cardiovascular Research Institute, National University of Singapore, Singapore (4) National University Heart Center, Singapore (5) High Tech Home Care AG, Rotkreuz, Switzerland (6) Department of Endocrinology and Clinical Nutrition, University Hospital Zurich and University of Zurich, 8091 Zürich, Switzerland

Bo Burla, PhD (Presenter) Sling @ National University of Singapore

Presenter Bio: I studied biology at the University of Zurich, Switzerland, and made my PhD in Molecular Plant Physiology in the lab of Prof. Enrico Martinoia, focusing on ABC transporters and comparative molecular phylogenetics. Subsequently, I became as researcher at the University Hospital of Zurich, where I was involved in establishing an LC/MS-based assay for the peptide hormone hepcidin and in projects studying Fabry Disease mechanisms. Biological processes, bioanalysis, clinical applications and the use of informatics to improve workflows have been my constant research interests. With this background I am now working as a senior researcher at the Singapore Lipidomics Incubator (SLING), heading our new data team that is focusing on developing workflows and software pipelines for the analytical data processing, QA/AC and exploration of the diverse lipidomics datasets generated by our lab.

Relevant Financial Disclosures (within past 24 months, reported on May 17, 2023) No relevant financial relationship(s) to disclose. Conflict mitigation NOT REQUIRED.

INTRODUCTION
Fabry disease (FD) is an X-linked lysosomal storage disorder characterized by reduced activity of α–galactosidase A (GLA) resulting in progressive accumulation of the globotriaosylceramide (Gb3). Clinical manifestations at later stages include chronic kidney disease, cardiomyopathies, and strokes. Estimated prevalence varies from 1:40,00 to 1:200,000 for classic FD, and down to 1:1,200 for later-onset FD. FD may also be present in initially unexplained diseases, e.g., cryptogenic stroke, cardiomyopathies, and chronic kidney disease.

In FD diagnosis, determining pathogenicity of detected variants may not always possible. In these cases, mass spectrometry (MS)–based analysis of plasma and urinary Gb3 and metabolites is recommended (PMID: 27195818). This indicates the need and utility of MS-based methods in screening and diagnosis of this under-diagnosed condition.

Platelets from a FD rat model were shown to have 7,000–fold increased Gb3 levels. In humans, a proinflammatory state and systemic inflammation have been reported for FD. However, no data is currently available on Gb3 profiles in human platelets from FD patients treated with enzyme-replacement therapy (ERT).

OBJECTIVES
The primary objective is to analyze Gb3 concentrations in platelets of ERT-treated FD patients and in healthy controls. The secondary objective was to test and validate previously reported Gb3 species (PMID: 23968398) in plasma of ERT-treated FD patients as diagnostic markers of FD. The third objective was conducting a comprehensive lipidomics analysis of both plasma and platelets from FD patients.

METHODS
Platelets were isolated using differential centrifugation in presence of anticoagulants. Plasma and platelet lipids were isolated via a simple single-step butanol/methanol liquid extraction.MS–based lipidomics analyses were performed using reversed-phase liquid chromatography and triplequadrupole MS.

RESULTS
Gb3 levels in platelets from ERT-treated FD patients were not increased compared to healthy controls. However, platelets exhibited high between-subject variability of Gb3, possibly associated with specific blood groups. Plasma Gb3 species harbouring sphingadiene (d18:2) showed an indeed better discrimination of FD patients from healthy controls compared to Gb3 species containing sphingosine (d18:1) that are commonly measured. The lipidomics profiling revealed changes in plasma GM3 gangliosides, sphingosine 1–phosphates, abundance of specific fatty acids in lipids, specific plasma ceramide ratios, and platelet acylcarnitines.

CONCLUSION
This study demonstrates that changes in plasma and platelets lipidomes in FD go beyond Gb3 and its metabolites. Furthermore, ERT seems effective in preventing Gb3 accumulation in platelets of FD patients. Lastly, this study supports the potential use of specific Gb3 species as novel makers for screening and diagnosis of FD.