Research Paper Advance Articles
Mitochondrial dysfunction, iron accumulation, lipid peroxidation, and inflammasome activation in cellular models derived from patients with multiple sclerosis
- 1 Centro Andaluz de Biología del Desarrollo (CABD-CSIC-Universidad Pablo de Olavide), Sevilla 41013, Spain
Received: September 10, 2024 Accepted: January 21, 2025 Published: February 6, 2025
https://doi.org/10.18632/aging.206198How to Cite
Copyright: © 2025 García-Salas et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system (CNS). Despite advancements in managing relapsing active illness, effective treatments for the irreversible progressive decline in MS remain limited.
Research employing skin fibroblasts obtained from patients with neurological disorders revealed modifications in cellular stress pathways and bioenergetics. However, research using MS patient-derived cellular models is scarce.
In this study, we collected fibroblasts from two MS patients to investigate cellular pathological alterations. We observed that MS fibroblasts showed a senescent morphology associated with iron/lipofuscin accumulation and altered expression of iron metabolism proteins. In addition, we found increased lipid peroxidation and downregulation of antioxidant enzymes expression levels in MS fibroblasts. When challenged against erastin, a ferroptosis inducer, MS fibroblasts showed decreased viability, suggesting increased sensitivity to ferroptosis. Furthermore, MS fibroblasts presented alterations in the expression levels of autophagy-related proteins. Interestingly, these alterations were associated with mitochondrial dysfunction and inflammasome activation. These findings were validated in 7 additional patient-derived cell lines.
Our findings suggest that the underlying stress phenotype of MS fibroblasts may be disease-specific and recapitulate the main cellular pathological alterations found in the disease such as mitochondrial dysfunction, iron accumulation, lipid peroxidation, inflammasome activation, and pro-inflammatory cytokine production.
Abbreviations
a.u.: arbitrary units; ARA70: Androgen Receptor Activator 70; Atg12: Autophagy-Related 12; Atg5: Autophagy-Related 5; ATM: Ataxia Telangiectasia Mutated; ATP5F1A: ATP Synthase F1 subunit alpha; BSA: Bovine Serum Albumine; CNS: central nervous system; COX IV: Cytochrome C Oxidase Subunit IV; CSF: cerebrospinal fluid; Def: deferiprone; DMEM: Dulbecco′s Modified Eagle Medium; DMSO: dimethyl sulfoxide; DMT1: Divalent Metal Transporter 1; EAE: experimental autoimmune encephalomyelitis; FBS: Fetal Bovine Serum; FCCP: carbonyl cyanide ptrifluoromethoxy-phenylhydrazone; Fe+2: ferrous iron; Fe+3: ferric iron; FTL: ferritin light chain; FXN: frataxin; GPX4: glutathione peroxidase 4; GSK3B: Glycogen Synthase Kinase 3 Beta; HBSS: Hank′s Balanced Salt Solution; HMGCR: 3-Hydroxy3-Methylglutaryl-Coenzyme A Reductase; HRP: horseradish peroxidase; ICP-MS: inductively coupled mass spectrometry; IL1B: interleukin 1-beta; IRP-1: Iron-responsive Element-Binding Protein 1; ISCU: iron sulfur cluster assembly enzyme; KLF2: Kruppel-Like Factor 2; LAMP1: Lysosomal Associated Membrane Protein 1; LC3B: Microtubule-Assocciated Protein 1A/1B Light Chain 3B; LIP: Labile Iron Pool; LYRM4: LYR motif-containing protein 4; MAPK1: Mitogen-Activated Protein Kinase 1; MnSOD: manganese superoxide dismutase; Mrfn2: mitoferrin 2; MRI: magnetic resonance imaging; MS: multiple sclerosis; mtCO2: Mitochondrially-Encoded Cytochrome C Oxidase subunit II; mtFTL: mitochondrial ferritin; NDUFA9: NADH:ubiquinone oxidoreductase subunit A9; NDUFS1: NADH:ubiquinone oxidoreductase core subunit S1; NFE2L1: Nuclear Factor, Erythroid 2 Like 1; NFS1: NFS1 cysteine desulfurase; NOX4: NAPDH oxidase 4; NRAS: Neuroblastoma Ras Vital Oncogene Homolog; NLRP3: NRL family pyrin domain containing 3; OCR: oxygen consumption rate; OXPHOS: oxidative phosphorylation; p62: Autophagy Receptor p62; PBS: phosphate-buffer saline; PCBP1: Poly(RC) Binding Protein 1; PFA: paraformaldehyde; PI: propidium iodide; PIK3CA: Phosphatidylinositol-4,5-Biphosphate 3-Kinase Catalytic Subunit Alpha; PMS: progressive multiple sclerosis; PPMS: primary-progressive multiple sclerosis; ROS: Reactive Oxygen Species; RPL8: Ribosomal Protein L8; RRMS: remitting-relapsing multiple sclerosis; RSL3: RAS-selective lethal 3; SOD1: superoxide dismutase 1; SPMS: secondary-progressive multiple sclerosis; Tf: transferrin; TfR: transferrin receptor; VDAC: voltage-dependent anion channel; VDAC3: voltage-dependent anion channel 3; VitE: vitamin E.