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Related Experiment Video

Updated: Jul 6, 2025

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Development of an LC-MS/MS Method to Measure Sphingolipids in CSF from Patients with Multiple Sclerosis.

Yadira X Perez-Paramo1, Dawn Dufield1, Rathna Veeramachaneni1

  • 1Department of Translational Medicine, Genentech, Inc., South San Francisco, California (Y.X.P.-P., C.H., A.R., R.C.W., W.R.M., V.G.A.); KCAS Bioanalytical Sciences, Olathe, Kansas (D.D., R.V., E.P.); Washington University School of Medicine, St Louis, Missouri (A.H.C.); University of California, San Francisco, California (J.M.G.); and Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania (A.B.-O.).

Molecular Pharmacology
|January 5, 2024
PubMed
Summary

Cerebrospinal fluid sphingolipids are elevated in multiple sclerosis patients. A new method reveals distinct lipid profiles in different MS types and shows treatment with ocrelizumab shifts these profiles, particularly in relapsing MS.

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Area of Science:

  • Neuroscience
  • Biochemistry
  • Analytical Chemistry

Background:

  • Multiple sclerosis (MS) is a central nervous system inflammatory disease with varied clinical courses.
  • Autoimmune responses in MS lead to demyelination and neuroaxonal damage.
  • Sphingolipids in cerebrospinal fluid (CSF) are potential biomarkers for MS activity.

Purpose of the Study:

  • To develop a sensitive, high-throughput method for quantifying sphingolipids in human CSF.
  • To investigate CSF sphingolipid profiles in patients with relapsing MS (RMS) and primary progressive MS (PPMS) compared to healthy controls.
  • To assess the impact of ocrelizumab treatment on CSF sphingolipid levels in MS patients.

Main Methods:

  • Development of a multiplexed liquid chromatography-tandem mass spectrometry assay.
  • Quantification of 14 sphingolipid species in CSF samples.
  • Analysis of CSF from healthy controls (n=10), PPMS patients (n=27), and RMS patients (n=17) before and after ocrelizumab treatment.

Main Results:

  • Median CSF sphingolipid levels were significantly higher in PPMS and RMS patients compared to healthy controls.
  • Ocrelizumab treatment led to an 8.6% decrease in overall sphingolipid levels in RMS patients after 52 weeks.
  • Specific sphingolipids, like C16 glucosylceramide and C18 ceramides, decreased in RMS patients post-treatment.
  • C16 glucosylceramide levels in PPMS patients correlated with CSF neurofilament heavy levels.

Conclusions:

  • CSF sphingolipid levels are altered in patients with multiple sclerosis.
  • Distinct sphingolipid signatures differentiate MS subtypes and respond to ocrelizumab therapy.
  • The developed method facilitates further research into sphingolipids as biomarkers for MS and other CNS disorders.