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Lipidomics and Functional Analyses of Platelets in Fabry Disease

Conditions
Fabry Disease
Registration Number
NCT02649660
Lead Sponsor
Spital Linth
Brief Summary

This study aims to evaluate whether platelets are biochemically and functionally altered in Fabry disease (FD) and therefore possibly implicated in FD manifestations such as cerebrovascular events. To test this hypothesis the investigators aim to compare platelet and plasma lipid profiles, as well as platelet function and coagulation parameters of FD patients and healthy controls.

Detailed Description

Fabry disease (FD) is a severe X-linked inborn error of the lysosomal glycosphingolipid metabolism. FD patients have significantly increased risks for cardiac and cerebrovascular events, which can also occur early and in absence of the typical FD symptoms. However, the pathophysiological mechanisms leading to vascular occlusion and ischemia in FD are largely unclear. Prevention of recurrent cerebrovascular events is usually based on empirical anti-platelet therapy.

Prothrombotic states and partially activated platelets have been reported for FD patients. Platelets contain glycosphingolipids, including globotriaosylceramide (Gb3), and have lysosomal α-galactosidase activity. To investigate whether the lack of or the reduced α-galactosidase enzyme activity present in Fabry disease affects platelet lipid metabolism the investigators plan to perform LC-MS-based lipidomics analyses of platelets and plasma in FD patients and healthy controls. To assess whether platelets are functionally altered in FD, the investigators aim to determine the activation status, activability, aggregability and other parameters along with plasma markers of coagulation using flow cytometry, aggregometry and immunoassays.

Recruitment & Eligibility

Status
UNKNOWN
Sex
All
Target Recruitment
32
Inclusion Criteria
  • Healthy Volunteers: without any known cardiovascular, cerebrovascular and renal diseases and without any known conditions affecting platelet function, blood coagulation and lipid metabolism.
  • Patients: Genetically confirmed Fabry Disease
  • Adult persons (18-65 years old), both female and male
  • Informed written consent
Exclusion Criteria
  • Failure to meet inclusion criteria
  • Pregnancy (as declared by the study participant, no pregnancy test will be performed)

Study & Design

Study Type
OBSERVATIONAL
Study Design
Not specified
Primary Outcome Measures
NameTimeMethod
Differences in sphingolipid profiles of platelets and plasma between Fabry disease patients and healthy subjectsBaseline

Determination of sphingolipid concentrations in isolated platelets and plasma by targeted LC-MS (liquid chromatography mass spectrometry)-based lipidomics analysis of globotriaosylceramides (Gb3), globotriaosylspingosines (Lyso-Gb3), globotetraosylceramides (Gb4), lactosylceramides (LacCer), glucosylceramides, ceramides, sphingomyelins, sphingosines and sphingosine-1-phosphates.

Differences in platelet function assessed by aggregometryBaseline

Determination of platelet aggregation after agonist stimulation with TRAP (thrombin receptor activator peptide), ADP (adenosine disphosphate) and arachidonic acid by light transmission aggregometry.

Secondary Outcome Measures
NameTimeMethod
Differences of alpha-galactosidase A enzyme activity in plasma and isolated platelets between Fabry disease patients and healthy subjectsBaseline

Analysis of the alpha-galactosidase A enzyme activity using a fluorometric enzyme assay with 4-methylumbelliferyl-α-D-galactopyranoside as substrate.

Differences in phospholipid profiles in platelets and plasma between Fabry disease patients and healthy subjectsBaseline

Determination of phospholipid concentrations in isolated platelets and plasma by targeted LC-MS-based lipidomics analysis of phosphatidylcholines, phosphatidylserines, phosphatidylinositols, and phosphatidylethanolamines.

Differences in plasma levels of the platelet activation marker sCD40L between Fabry disease patients and healthy subjectsBaseline

Plasma concentrations of sCD40L (soluble CD40 ligand) are determined by ELISA (enzyme-linked immunosorbent assay)

Differences in expression of the platelet activation marker P-selectin (CD62P) between Fabry disease patients and healthy subjects at baseline and after agonist stimulationBaseline

Flow cytometric analysis of platelet surface expression of CD62P in whole blood determined at baseline and after platelet activation with the agonists TRAP (thrombin receptor activator peptide), ADP, and arachidonic acid.

Differences in plasma levels of the platelet activation marker soluble P-selectin between Fabry disease patients and healthy subjectsBaseline

Plasma concentrations of soluble P-selectin (soluble CD62P) are determined by ELISA (enzyme-linked immunosorbent assay)

Differences in presence of platelet aggregates between Fabry disease patients and healthy subjectsBaseline

Flow cytometric assessment of platelet-platelet and platelet-leukocyte aggregates

Differences in expression of the platelet activation marker CD63 between Fabry disease patients and healthy subjects at baseline and after agonist stimulationBaseline

Flow cytometric analysis of platelet surface expression of CD63 in whole blood determined at baseline and after platelet activation with the agonists TRAP (thrombin receptor activator peptide), ADP, and arachidonic acid.

Trial Locations

Locations (2)

University Hospital, Zürich

🇨🇭

Zürich, ZH, Switzerland

Spital Linth

🇨🇭

Uznach, SG, Switzerland

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