Bioenergetic and Autophagic Characterization of Skin Fibroblasts from C9orf72 Patients
Abstract
:1. Introduction
2. Methods
2.1. Subjects
2.2. Fibroblasts Cell Culture
2.3. C9ORF72 and SUMO2/3 Protein Quantification
2.4. Oxidative Stress Assessment
2.5. Mitochondrial Metabolism Assay
2.6. Autophagic Flux Analysis
2.7. Functional Enrichment Analysis
2.8. Accession of Data
2.9. Statistical Analysis
3. Results
3.1. Fibroblasts from C9ALS/FTD Patients Show Reduced C9orf72 Protein Levels
3.2. Fibroblasts from C9ALS/FTD Patients Evidence Increased Levels of Oxidative Stress
3.3. Fibroblasts from C9ALS/FTD Patients Evidence Mitochondrial Dysfunction and Altered Metabolism
3.4. Fibroblasts from C9ALS/FTD Patients Show Increased SUMO2/3 Protein Quantification
3.5. Autophagy- and SUMOylation-Related Pathways Are Significantly Altered in Fibroblasts from C9ALS/FTD Patients
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sex | Age (Years) | G4C2 | Clinical Diagnosis | |
---|---|---|---|---|
C9ORF72_1 | Female | 70 | 2, >145 | FTD |
C9ORF72_2 | Male | 76 | 2, >145 | ALS/FTD |
C9ORF72_3 | Female | 65 | 2, >145 | ALS/FTD |
C9ORF72_4 | Female | 69 | 2, >145 | FTD |
Control_1 | Female | 82 | 2, 2 | Control |
Control_2 | Male | 57 | 2, 6 | Control |
Control_3 | Male | 86 | 2, 4 | Control |
Control_4 | Female | 68 | 2, 2 | Control |
Enzyme Activities 1 | Fibroblasts | ||||
C9ORF72_1 | C9ORF72_2 | C9ORF72_3 | C9ORF72_4 | Control Range | |
Complex I+III | 17.4 | 9.6 * | 0 * | 25.6 | 15–42 |
Complex II | 19.2 * | 34 | 18.1 * | 40.6 | 22–35 |
Complex II+III | 3.4 * | 9.8 * | 1.8 * | 15.8 | 11–20 |
Complex III | 4.6 * | 8.3 * | 0 * | 14.2 * | 25–48 |
Complex IV | 67 | 42.2 | 18.1 * | 68.5 | 33–57 |
Citrate synthetase (CS) | 18.7 * | 54 | 15.8 * | 61.8 | 41–62 |
Enzyme Activities 2 | C9ORF72_1 | C9ORF72_2 | C9ORF72_3 | C9ORF72_4 | Control Range |
Complex I+III/CS | 0.930 | 0.178 * | 0 * | 0.414 | 0.24–0.85 |
Complex II/CS | 1.027 | 0.630 | 1.146 | 0.657 | 0.53–0.75 |
Complex II+III/CS | 0.182 * | 0.181 * | 0.114 * | 0.256 | 0.25–0.42 |
Complex III/CS | 0.246 * | 0.154 * | 0 * | 0.230 * | 0.49–1.06 |
Complex IV/CS | 3.583 | 0.781 | 1.146 | 1.108 | 0.84–1.15 |
PATH_ID | PATH_NAME | Size | padj | LOR |
---|---|---|---|---|
R-HSA-1632852 | Macroautophagy | 120 | 0.00004267 | −0.4304 |
R-HSA-9612973 | Autophagy | 132 | 0.0002208 | −0.3759 |
R-HSA-9663891 | Selective autophagy | 67 | 0.0005462 | −0.4965 |
R-HSA-3108232 | SUMO E3 ligases SUMOylate target proteins | 151 | 0.000003633 | 0.4239 |
R-HSA-2990846 | SUMOylation | 157 | 0.000006105 | 0.4079 |
R-HSA-983168 | Antigen processing: Ubiquitination and Proteasome degradation | 284 | 0.004528 | −0.2058 |
R-HSA-8852135 | Protein ubiquitination | 63 | 0.005484 | −0.4211 |
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Alvarez-Mora, M.I.; Garrabou, G.; Barcos, T.; Garcia-Garcia, F.; Grillo-Risco, R.; Peruga, E.; Gort, L.; Borrego-Écija, S.; Sanchez-Valle, R.; Canto-Santos, J.; et al. Bioenergetic and Autophagic Characterization of Skin Fibroblasts from C9orf72 Patients. Antioxidants 2022, 11, 1129. https://doi.org/10.3390/antiox11061129
Alvarez-Mora MI, Garrabou G, Barcos T, Garcia-Garcia F, Grillo-Risco R, Peruga E, Gort L, Borrego-Écija S, Sanchez-Valle R, Canto-Santos J, et al. Bioenergetic and Autophagic Characterization of Skin Fibroblasts from C9orf72 Patients. Antioxidants. 2022; 11(6):1129. https://doi.org/10.3390/antiox11061129
Chicago/Turabian StyleAlvarez-Mora, Maria Isabel, Gloria Garrabou, Tamara Barcos, Francisco Garcia-Garcia, Ruben Grillo-Risco, Emma Peruga, Laura Gort, Sergi Borrego-Écija, Raquel Sanchez-Valle, Judith Canto-Santos, and et al. 2022. "Bioenergetic and Autophagic Characterization of Skin Fibroblasts from C9orf72 Patients" Antioxidants 11, no. 6: 1129. https://doi.org/10.3390/antiox11061129