Mutant WDR45 Leads to Altered Ferritinophagy and Ferroptosis in β-Propeller Protein-Associated Neurodegeneration
Abstract
:1. Introduction
2. Results
2.1. Clinical Presentation and Genetic Analysis of Patient 1 Harboring a Recurrent Mutation in WDR45
2.2. WDR45-Mutant Fibroblasts Exhibit Diminished Lysosomal Integrity and Altered Mitochondrial Network
2.3. WDR45-Mutant Fibroblasts Exhibit Altered Autophagy
2.4. Loss of WDR45 Is Linked to Disrupted Iron Recycling
3. Discussion
4. Materials and Methods
4.1. Subjects
4.2. Cell Culture
4.3. Sanger Sequencing and Quantitative PCR Analysis
4.4. Western Blot Analysis
4.5. Immunofluorescence Staining and Image Analysis
4.6. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Gregory, A.; Hayflick, S. Neurodegeneration with Brain Iron Accumulation Disorders Overview. In GeneReviews®; Adam, M.P., Ardinger, H.H., Pagon, R.A., Wallace, S.E., Bean, L.J., Gripp, K.W., Mirzaa, G.M., Amemiya, A., Eds.; University of Washington: Seattle, WA, USA, 1993. [Google Scholar]
- Saitsu, H.; Nishimura, T.; Muramatsu, K.; Kodera, H.; Kumada, S.; Sugai, K.; Kasai-Yoshida, E.; Sawaura, N.; Nishida, H.; Hoshino, A.; et al. De Novo Mutations in the Autophagy Gene WDR45 Cause Static Encephalopathy of Childhood with Neurodegeneration in Adulthood. Nat. Genet. 2013, 45, 445–449. [Google Scholar] [CrossRef] [PubMed]
- Haack, T.B.; Hogarth, P.; Kruer, M.C.; Gregory, A.; Wieland, T.; Schwarzmayr, T.; Graf, E.; Sanford, L.; Meyer, E.; Kara, E.; et al. Exome Sequencing Reveals De Novo WDR45 Mutations Causing a Phenotypically Distinct, X-Linked Dominant Form of NBIA. Am. J. Hum. Genet. 2012, 91, 1144–1149. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hayflick, S.J.; Kruer, M.C.; Gregory, A.; Haack, T.B.; Kurian, M.A.; Houlden, H.H.; Anderson, J.; Boddaert, N.; Sanford, L.; Harik, S.I.; et al. Beta-Propeller Protein-Associated Neurodegeneration: A New X-Linked Dominant Disorder with Brain Iron Accumulation. Brain 2013, 136, 1708–1717. [Google Scholar] [CrossRef] [PubMed]
- Arber, C.E.; Li, A.; Houlden, H.; Wray, S. Review: Insights into Molecular Mechanisms of Disease in Neurodegeneration with Brain Iron Accumulation: Unifying Theories: Mechanisms of Neurodegeneration with Brain Iron Accumulation. Neuropathol. Appl. Neurobiol. 2016, 42, 220–241. [Google Scholar] [CrossRef]
- Zhao, Y.G.; Sun, L.; Miao, G.; Ji, C.; Zhao, H.; Sun, H.; Miao, L.; Yoshii, S.R.; Mizushima, N.; Wang, X.; et al. The Autophagy Gene Wdr45/Wipi4 Regulates Learning and Memory Function and Axonal Homeostasis. Autophagy 2015, 11, 881–890. [Google Scholar] [CrossRef] [Green Version]
- Bakula, D.; Müller, A.J.; Zuleger, T.; Takacs, Z.; Franz-Wachtel, M.; Thost, A.-K.; Brigger, D.; Tschan, M.P.; Frickey, T.; Robenek, H.; et al. WIPI3 and WIPI4 β-Propellers Are Scaffolds for LKB1-AMPK-TSC Signalling Circuits in the Control of Autophagy. Nat. Commun. 2017, 8, 15637. [Google Scholar] [CrossRef]
- Wan, H.; Wang, Q.; Chen, X.; Zeng, Q.; Shao, Y.; Fang, H.; Liao, X.; Li, H.-S.; Liu, M.-G.; Xu, T.-L.; et al. WDR45 Contributes to Neurodegeneration through Regulation of ER Homeostasis and Neuronal Death. Autophagy 2020, 16, 531–547. [Google Scholar] [CrossRef]
- Seibler, P.; Burbulla, L.F.; Dulovic, M.; Zittel, S.; Heine, J.; Schmidt, T.; Rudolph, F.; Westenberger, A.; Rakovic, A.; Münchau, A.; et al. Iron Overload Is Accompanied by Mitochondrial and Lysosomal Dysfunction in WDR45 Mutant Cells. Brain 2018, 141, 3052–3064. [Google Scholar] [CrossRef]
- Arosio, P.; Elia, L.; Poli, M. Ferritin, Cellular Iron Storage and Regulation. IUBMB Life 2017, 69, 414–422. [Google Scholar] [CrossRef]
- Mancias, J.D.; Wang, X.; Gygi, S.P.; Harper, J.W.; Kimmelman, A.C. Quantitative Proteomics Identifies NCOA4 as the Cargo Receptor Mediating Ferritinophagy. Nature 2014, 509, 105–109. [Google Scholar] [CrossRef]
- Quiles del Rey, M.; Mancias, J.D. NCOA4-Mediated Ferritinophagy: A Potential Link to Neurodegeneration. Front. Neurosci. 2019, 13, 238. [Google Scholar] [CrossRef] [Green Version]
- Mancias, J.D.; Pontano Vaites, L.; Nissim, S.; Biancur, D.E.; Kim, A.J.; Wang, X.; Liu, Y.; Goessling, W.; Kimmelman, A.C.; Harper, J.W. Ferritinophagy via NCOA4 Is Required for Erythropoiesis and Is Regulated by Iron Dependent HERC2-Mediated Proteolysis. eLife 2015, 4, e10308. [Google Scholar] [CrossRef] [PubMed]
- Dixon, S.J.; Lemberg, K.M.; Lamprecht, M.R.; Skouta, R.; Zaitsev, E.M.; Gleason, C.E.; Patel, D.N.; Bauer, A.J.; Cantley, A.M.; Yang, W.S.; et al. Ferroptosis: An Iron-Dependent Form of Nonapoptotic Cell Death. Cell 2012, 149, 1060–1072. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ajoolabady, A.; Aslkhodapasandhokmabad, H.; Libby, P.; Tuomilehto, J.; Lip, G.Y.H.; Penninger, J.M.; Richardson, D.R.; Tang, D.; Zhou, H.; Wang, S.; et al. Ferritinophagy and Ferroptosis in the Management of Metabolic Diseases. Trends Endocrinol. Metab. 2021, 32, 444–462. [Google Scholar] [CrossRef]
- Latunde-Dada, G.O. Ferroptosis: Role of Lipid Peroxidation, Iron and Ferritinophagy. Biochim. Biophys. Acta BBA-Gen. Subj. 2017, 1861, 1893–1900. [Google Scholar] [CrossRef] [Green Version]
- Ndayisaba, A.; Kaindlstorfer, C.; Wenning, G.K. Iron in Neurodegeneration—Cause or Consequence? Front. Neurosci. 2019, 13, 180. [Google Scholar] [CrossRef] [Green Version]
- Boag, M.K.; Roberts, A.; Uversky, V.N.; Ma, L.; Richardson, D.R.; Pountney, D.L. Ferritinophagy and α-Synuclein: Pharmacological Targeting of Autophagy to Restore Iron Regulation in Parkinson’s Disease. Int. J. Mol. Sci. 2022, 23, 2378. [Google Scholar] [CrossRef] [PubMed]
- Richards, S.; Aziz, N.; Bale, S.; Bick, D.; Das, S.; Gastier-Foster, J.; Grody, W.W.; Hegde, M.; Lyon, E.; Spector, E.; et al. Standards and Guidelines for the Interpretation of Sequence Variants: A Joint Consensus Recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet. Med. 2015, 17, 405–424. [Google Scholar] [CrossRef] [Green Version]
- Kurz, T.; Terman, A.; Gustafsson, B.; Brunk, U.T. Lysosomes in Iron Metabolism, Ageing and Apoptosis. Histochem. Cell Biol. 2008, 129, 389–406. [Google Scholar] [CrossRef] [Green Version]
- Kalia, L.V.; Lang, A.E. Parkinson’s Disease. Lancet 2015, 386, 896–912. [Google Scholar] [CrossRef]
- Montalvo, A.L.E.; Bembi, B.; Donnarumma, M.; Filocamo, M.; Parenti, G.; Rossi, M.; Merlini, L.; Buratti, E.; De Filippi, P.; Dardis, A.; et al. Mutation Profile of TheGAA Gene in 40 Italian Patients with Late Onset Glycogen Storage Disease Type II. Hum. Mutat. 2006, 27, 999–1006. [Google Scholar] [CrossRef] [PubMed]
- Li, H.; Ham, A.; Ma, T.C.; Kuo, S.-H.; Kanter, E.; Kim, D.; Ko, H.S.; Quan, Y.; Sardi, S.P.; Li, A.; et al. Mitochondrial Dysfunction and Mitophagy Defect Triggered by Heterozygous GBA Mutations. Autophagy 2019, 15, 113–130. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Seiler, A.; Schneider, M.; Förster, H.; Roth, S.; Wirth, E.K.; Culmsee, C.; Plesnila, N.; Kremmer, E.; Rådmark, O.; Wurst, W.; et al. Glutathione Peroxidase 4 Senses and Translates Oxidative Stress into 12/15-Lipoxygenase Dependent- and AIF-Mediated Cell Death. Cell Metab. 2008, 8, 237–248. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yang, W.S.; SriRamaratnam, R.; Welsch, M.E.; Shimada, K.; Skouta, R.; Viswanathan, V.S.; Cheah, J.H.; Clemons, P.A.; Shamji, A.F.; Clish, C.B.; et al. Regulation of Ferroptotic Cancer Cell Death by GPX4. Cell 2014, 156, 317–331. [Google Scholar] [CrossRef] [Green Version]
- Vučković, A.-M.; Bosello Travain, V.; Bordin, L.; Cozza, G.; Miotto, G.; Rossetto, M.; Toppo, S.; Venerando, R.; Zaccarin, M.; Maiorino, M.; et al. Inactivation of the Glutathione Peroxidase GPx4 by the Ferroptosis-Inducing Molecule RSL3 Requires the Adaptor Protein 14-3-3ε. FEBS Lett. 2020, 594, 611–624. [Google Scholar] [CrossRef]
- Imai, H.; Matsuoka, M.; Kumagai, T.; Sakamoto, T.; Koumura, T. Lipid Peroxidation-Dependent Cell Death Regulated by GPx4 and Ferroptosis. In Apoptotic and Non-Apoptotic Cell Death; Current Topics in Microbiology and, Immunology; Nagata, S., Nakano, H., Eds.; Springer International Publishing: Cham, Switzerland, 2017; pp. 143–170. ISBN 978-3-319-23913-2. [Google Scholar]
- Pankiv, S.; Clausen, T.H.; Lamark, T.; Brech, A.; Bruun, J.-A.; Outzen, H.; Øvervatn, A.; Bjørkøy, G.; Johansen, T. P62/SQSTM1 Binds Directly to Atg8/LC3 to Facilitate Degradation of Ubiquitinated Protein Aggregates by Autophagy. J. Biol. Chem. 2007, 282, 24131–24145. [Google Scholar] [CrossRef] [Green Version]
- Bjørkøy, G.; Lamark, T.; Brech, A.; Outzen, H.; Perander, M.; Øvervatn, A.; Stenmark, H.; Johansen, T. P62/SQSTM1 Forms Protein Aggregates Degraded by Autophagy and Has a Protective Effect on Huntingtin-Induced Cell Death. J. Cell Biol. 2005, 171, 603–614. [Google Scholar] [CrossRef] [Green Version]
- Sun, Y.; He, L.; Wang, T.; Hua, W.; Qin, H.; Wang, J.; Wang, L.; Gu, W.; Li, T.; Li, N.; et al. Activation of P62-Keap1-Nrf2 Pathway Protects 6-Hydroxydopamine-Induced Ferroptosis in Dopaminergic Cells. Mol. Neurobiol. 2020, 57, 4628–4641. [Google Scholar] [CrossRef]
- Ichimura, Y.; Waguri, S.; Sou, Y.-S.; Kageyama, S.; Hasegawa, J.; Ishimura, R.; Saito, T.; Yang, Y.; Kouno, T.; Fukutomi, T.; et al. Phosphorylation of P62 Activates the Keap1-Nrf2 Pathway during Selective Autophagy. Mol. Cell 2013, 51, 618–631. [Google Scholar] [CrossRef] [Green Version]
- Appelqvist, H.; Wäster, P.; Kågedal, K.; Öllinger, K. The Lysosome: From Waste Bag to Potential Therapeutic Target. J. Mol. Cell Biol. 2013, 5, 214–226. [Google Scholar] [CrossRef] [Green Version]
- Urrutia, P.; Mena, N.; Nunez, M. The Interplay between Iron Accumulation, Mitochondrial Dysfunction, and Inflammation during the Execution Step of Neurodegenerative Disorders. Front. Pharmacol. 2014, 5, 38. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hor, C.H.H.; Tang, B.L. Beta-Propeller Protein-Associated Neurodegeneration (BPAN) as a Genetically Simple Model of Multifaceted Neuropathology Resulting from Defects in Autophagy. Rev. Neurosci. 2019, 30, 261–277. [Google Scholar] [CrossRef] [PubMed]
- Li, C.; Deng, X.; Xie, X.; Liu, Y.; Friedmann Angeli, J.P.; Lai, L. Activation of Glutathione Peroxidase 4 as a Novel Anti-Inflammatory Strategy. Front. Pharmacol. 2018, 9, 1120. [Google Scholar] [CrossRef] [PubMed]
- Conrad, M.; Friedmann Angeli, J.P. Glutathione Peroxidase 4 (Gpx4) and Ferroptosis: What’s so Special about It? Mol. Cell. Oncol. 2015, 2, e995047. [Google Scholar] [CrossRef] [Green Version]
- Ursini, F.; Maiorino, M.; Valente, M.; Ferri, L.; Gregolin, C. Purification from Pig Liver of a Protein Which Protects Liposomes and Biomembranes from Peroxidative Degradation and Exhibits Glutathione Peroxidase Activity on Phosphatidylcholine Hydroperoxides. Biochim. Biophys. Acta BBA-Lipids Lipid Metab. 1982, 710, 197–211. [Google Scholar] [CrossRef]
- Thomas, J.P.; Geiger, P.G.; Maiorino, M.; Ursini, F.; Girotti, A.W. Enzymatic Reduction of Phospholipid and Cholesterol Hydroperoxides in Artificial Bilayers and Lipoproteins. Biochim. Biophys. Acta BBA-Lipids Lipid Metab. 1990, 1045, 252–260. [Google Scholar] [CrossRef]
- Brigelius-Flohé, R.; Maiorino, M. Glutathione Peroxidases. Biochim. Biophys. Acta 2013, 1830, 3289–3303. [Google Scholar] [CrossRef]
- Angeli, J.P.F.; Schneider, M.; Proneth, B.; Tyurina, Y.Y.; Tyurin, V.A.; Hammond, V.J.; Herbach, N.; Aichler, M.; Walch, A.; Eggenhofer, E.; et al. Inactivation of the Ferroptosis Regulator Gpx4 Triggers Acute Renal Failure in Mice. Nat. Cell Biol. 2014, 16, 1180–1191. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xiong, Q.; Li, X.; Li, W.; Chen, G.; Xiao, H.; Li, P.; Wu, C. WDR45 Mutation Impairs the Autophagic Degradation of Transferrin Receptor and Promotes Ferroptosis. Front. Mol. Biosci. 2021, 8, 645831. [Google Scholar] [CrossRef]
- Yang, W.S.; Stockwell, B.R. Ferroptosis: Death by Lipid Peroxidation. Trends Cell Biol. 2016, 26, 165–176. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Goodwin, J.M.; Dowdle, W.E.; DeJesus, R.; Wang, Z.; Bergman, P.; Kobylarz, M.; Lindeman, A.; Xavier, R.J.; McAllister, G.; Nyfeler, B.; et al. Autophagy-Independent Lysosomal Targeting Regulated by ULK1/2-FIP200 and ATG9. Cell Rep. 2017, 20, 2341–2356. [Google Scholar] [CrossRef] [Green Version]
- Grünewald, A.; Voges, L.; Rakovic, A.; Kasten, M.; Vandebona, H.; Hemmelmann, C.; Lohmann, K.; Orolicki, S.; Ramirez, A.; Schapira, A.H.V.; et al. Mutant Parkin Impairs Mitochondrial Function and Morphology in Human Fibroblasts. PLoS ONE 2010, 5, e12962. [Google Scholar] [CrossRef]
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Diaw, S.H.; Ganos, C.; Zittel, S.; Plötze-Martin, K.; Kulikovskaja, L.; Vos, M.; Westenberger, A.; Rakovic, A.; Lohmann, K.; Dulovic-Mahlow, M. Mutant WDR45 Leads to Altered Ferritinophagy and Ferroptosis in β-Propeller Protein-Associated Neurodegeneration. Int. J. Mol. Sci. 2022, 23, 9524. https://doi.org/10.3390/ijms23179524
Diaw SH, Ganos C, Zittel S, Plötze-Martin K, Kulikovskaja L, Vos M, Westenberger A, Rakovic A, Lohmann K, Dulovic-Mahlow M. Mutant WDR45 Leads to Altered Ferritinophagy and Ferroptosis in β-Propeller Protein-Associated Neurodegeneration. International Journal of Molecular Sciences. 2022; 23(17):9524. https://doi.org/10.3390/ijms23179524
Chicago/Turabian StyleDiaw, Sokhna Haissatou, Christos Ganos, Simone Zittel, Kirstin Plötze-Martin, Leonora Kulikovskaja, Melissa Vos, Ana Westenberger, Aleksandar Rakovic, Katja Lohmann, and Marija Dulovic-Mahlow. 2022. "Mutant WDR45 Leads to Altered Ferritinophagy and Ferroptosis in β-Propeller Protein-Associated Neurodegeneration" International Journal of Molecular Sciences 23, no. 17: 9524. https://doi.org/10.3390/ijms23179524