Abstract
Amyotrophic lateral sclerosis (ALS) is characterized by a wide metabolic remodeling, as shown by recent metabolomics and lipidomics studies performed in samples from patient cohorts and experimental animal models. Here, we explored the metabolome and lipidome of fibroblasts from sporadic ALS patients (n = 13) comparatively to age- and sex-matched controls (n = 11), and the subcellular fraction containing the mitochondria and endoplasmic reticulum (mito-ER), given that mitochondrial dysfunctions and ER stress are important features of ALS patho-mechanisms. We also assessed the mitochondrial oxidative respiration and the mitochondrial genomic (mtDNA) sequence, although without yielding significant differences. Compared to controls, ALS fibroblasts did not exhibit a mitochondrial respiration defect nor an increased proportion of mitochondrial DNA mutations. In addition, non-targeted metabolomics and lipidomics analyses identified 124 and 127 metabolites, and 328 and 220 lipids in whole cells and the mito-ER fractions, respectively, along with partial least-squares–discriminant analysis (PLS-DA) models being systematically highly predictive of the disease. The most discriminant metabolomic features were the alteration of purine, pyrimidine, and energetic metabolisms, suggestive of oxidative stress and of pro-inflammatory status. The most important lipidomic feature in the mito-ER fraction was the disturbance of phosphatidylcholine PC (36:4p) levels, which we had previously reported in the cerebrospinal fluid of ALS patients and in the brain from an ALS mouse model. Thus, our results reveal that fibroblasts from sporadic ALS patients share common metabolic remodeling, consistent with other metabolic studies performed in ALS, opening perspectives for further exploration in this cellular model in ALS.
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Abbreviations
- ALS:
-
amyotrophic lateral sclerosis
- sALS:
-
sporadic ALS
- ER:
-
endoplasmic reticulum
- mtDNA:
-
mitochondrial DNA
- PCA:
-
principal component analysis
- PLS-DA:
-
partial least square discriminant analysis
- OXPHOS:
-
oxidative phosphorylation
- FVC:
-
forced vital capacity
- BMI:
-
body mass index
- HRMS:
-
high-resolution mass spectrometry
- UPLC:
-
ultra-performance liquid chromatography
- ESI:
-
electrospray ionization
- QC:
-
quality control
- VIP:
-
variable importance in projection
- OCR:
-
oxygen consumption rate
- RCR:
-
respiration capacity rate
- SM:
-
sphingomyelins
- PC:
-
phosphatidylcholines
- PE:
-
phosphatidylethanolamines
- ROS:
-
reactive oxygen species
- CSF:
-
cerebrospinal fluid
- Cer:
-
ceramides
- DHA:
-
docosahexaenoic acid
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We thank University Hospitals of Angers and Limoges for patients’ recruitment and fibroblasts’ sampling.
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C.V-D and H.B designed and supervised the study, performed statistical analysis and wrote the manuscript; C. Br performed mtDNA experiments and analyzed mtDNA sequence; C.V-D and C.Bo performed metabolomic and lipidomic assays, S.C performed cell cultures and mitochondrial function assessment; P.Cod and B.F recruited ALS patients and performed fibroblast sampling; J.C and P. Cou supervised patients’ recruitment; P.V supervised genetic determination of genes involved in ALS; R.H gave technical and intellectual support and critical advice for English writing; P.Cor, P.V, and C.R.A gave technical and intellectual support; G.L and P.R gave intellectual support and conceptual advice for manuscript writing. All authors offered conceptual advice and comments on the manuscript. All authors read and approved the final manuscript.
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All the participants in this current study provided their informed consent for the use of their fibroblasts for research. The ethic committees of the Centre for Human Research of Angers and Limoges Hospitals approved the study and the consent process.
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Veyrat-Durebex, C., Bris, C., Codron, P. et al. Metabo-lipidomics of Fibroblasts and Mitochondrial-Endoplasmic Reticulum Extracts from ALS Patients Shows Alterations in Purine, Pyrimidine, Energetic, and Phospholipid Metabolisms. Mol Neurobiol 56, 5780–5791 (2019). https://doi.org/10.1007/s12035-019-1484-7
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DOI: https://doi.org/10.1007/s12035-019-1484-7