Metabolomic Profiling of Cerebral Palsy Brain Tissue Reveals Novel Central Biomarkers and Biochemical Pathways Associated with the Disease: A Pilot Study

Abstract

Cerebral palsy (CP) is one of the most common causes of motor disability in childhood, with complex and heterogeneous etiopathophysiology and clinical presentation. Understanding the metabolic processes associated with the disease may aid in the discovery of preventive measures and therapy. Tissue samples (caudate nucleus) were obtained from post-mortem CP cases (n = 9) and age- and gender-matched control subjects (n = 11). We employed a targeted metabolomics approach using both 1H NMR and direct injection liquid chromatography-tandem mass spectrometry (DI/LC-MS/MS). We accurately identified and quantified 55 metabolites using 1H NMR and 186 using DI/LC-MS/MS. Among the 222 detected metabolites, 27 showed significant concentration changes between CP cases and controls. Glycerophospholipids and urea were the most commonly selected metabolites used to develop predictive models capable of discriminating between CP and controls. Metabolomics enrichment analysis identified folate, propanoate, and androgen/estrogen metabolism as the top three significantly perturbed pathways. We report for the first time the metabolomic profiling of post-mortem brain tissue from patients who died from cerebral palsy. These findings could help to further investigate the complex etiopathophysiology of CP while identifying predictive, central biomarkers of CP.

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“…The samples were ultrasonicated 10 min in ice and vortexed for 1 min. Proteins and other impurities were separated by centrifugation at 13,000× g for 15 min at 4 • C. The supernatant was collected and 10 µL was used for analysis by DI/LC-MS/MS [65].…”

“…The singlet produced by the DSS methyl groups was used as an internal standard for chemical shift referencing (set to 0 ppm, concentration 1000 µM) and for quantification. All spectra were processed and analyzed using Chenomx NMR Suite (v8.1, Edmonton, AB, Canada) [65].…”

“…Then, 200 µL of the reconstituted sample was transferred to a 3 mm Bruker NMR tube for analysis. All samples were housed at 4 • C in a thermostatically controlled SampleJet autosampler (Bruker-Biospin, Billerica, MA, USA) and heated to room temperature over 3 min prior to analysis by NMR[65].…”

Breath indeed carries significant information about a disease: Potential biomarkers of cerebral palsy

“…The former is not surprising, as PPA is generated in the gut, but the latter should be clarified. PPA generated in the gut and stored in areas of damage within the brain's white matter [30] can proof two realistic scenarios. The first scenario implies high PPA and PPAE production in the gut, with the background of normal permeability of BBB and GBB in the case of CP.…”

“…Let us now review the results of another research group conducted morphological analysis in the CP case. In a recent study [30], a postmortem brain biopsy was analyzed by using mass spectrometry and nuclear magnetic resonance spectroscopy. A set of metabolites revealed by both techniques, contains PPA as the second most frequent specific metabolite on the list.…”