Acute manganese treatment restores defective autophagic cargo loading in Huntington's disease cell lines

Miles R. Bryan, Michael T. O'Brien, Kristen D. Nordham, Daniel I.R. Rose, Audra M. Foshage, Piyush Joshi, Rachana Nitin, Michael A. Uhouse, Alba Di Pardo, Ziyan Zhang, Vittorio Maglione, Michael Aschner, Aaron B. Bowman

Research output: Contribution to journalArticle

Abstract

The molecular etiology linking the pathogenic mutations in the Huntingtin (Htt) gene with Huntington's disease (HD) is unknown. Prior work suggests a role for Htt in neuronal autophagic function and mutant HTT protein disrupts autophagic cargo loading. Reductions in the bioavailability of the essential metal manganese (Mn) are seen in models of HD. Excess cellular Mn impacts autophagic function, but the target and molecular basis of these changes are unknown. Thus, we sought to determine if changes in cellular Mn status impact autophagic processes in a wild-type or mutant Htt-dependent manner. We report that the HD genotype is associated with reduced Mn-induced autophagy and that acute Mn exposure increases autophagosome induction/formation. To determine if a deficit in bioavailable Mn is mechanistically linked to the autophagy-related HD cellular phenotypes, we examined autophagosomes by electron microscopy. We observed that a 24 h 100 uM Mn restoration treatment protocol attenuated an established HD 'cargo-recognition failure' in the STHdh HD model cells by increasing the percentage of filled autophagosomes. Mn restoration had no effect on HTT aggregate number, but a 72 h co-treatment with chloroquine (CQ) in GFP-72Q-expressing HEK293 cells increased the number of visible aggregates in a dose-dependent manner. As CQ prevents autophagic degradation this indicates that Mn restoration in HD cell models facilitates incorporation of aggregates into autophagosomes. Together, these findings suggest that defective Mn homeostasis in HD models is upstream of the impaired autophagic flux and provide proof-of-principle support for increasing bioavailable Mn in HD to restore autophagic function and promote aggregate clearance.

Original languageEnglish (US)
Pages (from-to)3825-3841
Number of pages17
JournalHuman molecular genetics
Volume28
Issue number22
DOIs
StatePublished - Nov 15 2019

Fingerprint

Huntington Disease
Manganese
Cell Line
Therapeutics
Autophagy
Chloroquine
HEK293 Cells
Mutant Proteins
Clinical Protocols
Biological Availability
Electron Microscopy
Homeostasis
Cell Count
Metals
Genotype
Phenotype

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

Cite this

Bryan, M. R., O'Brien, M. T., Nordham, K. D., Rose, D. I. R., Foshage, A. M., Joshi, P., ... Bowman, A. B. (2019). Acute manganese treatment restores defective autophagic cargo loading in Huntington's disease cell lines. Human molecular genetics, 28(22), 3825-3841. https://doi.org/10.1093/hmg/ddz209

Acute manganese treatment restores defective autophagic cargo loading in Huntington's disease cell lines. / Bryan, Miles R.; O'Brien, Michael T.; Nordham, Kristen D.; Rose, Daniel I.R.; Foshage, Audra M.; Joshi, Piyush; Nitin, Rachana; Uhouse, Michael A.; Di Pardo, Alba; Zhang, Ziyan; Maglione, Vittorio; Aschner, Michael; Bowman, Aaron B.

In: Human molecular genetics, Vol. 28, No. 22, 15.11.2019, p. 3825-3841.

Research output: Contribution to journalArticle

Bryan, MR, O'Brien, MT, Nordham, KD, Rose, DIR, Foshage, AM, Joshi, P, Nitin, R, Uhouse, MA, Di Pardo, A, Zhang, Z, Maglione, V, Aschner, M & Bowman, AB 2019, 'Acute manganese treatment restores defective autophagic cargo loading in Huntington's disease cell lines', Human molecular genetics, vol. 28, no. 22, pp. 3825-3841. https://doi.org/10.1093/hmg/ddz209
Bryan, Miles R. ; O'Brien, Michael T. ; Nordham, Kristen D. ; Rose, Daniel I.R. ; Foshage, Audra M. ; Joshi, Piyush ; Nitin, Rachana ; Uhouse, Michael A. ; Di Pardo, Alba ; Zhang, Ziyan ; Maglione, Vittorio ; Aschner, Michael ; Bowman, Aaron B. / Acute manganese treatment restores defective autophagic cargo loading in Huntington's disease cell lines. In: Human molecular genetics. 2019 ; Vol. 28, No. 22. pp. 3825-3841.
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AU - Bryan, Miles R.

AU - O'Brien, Michael T.

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AU - Rose, Daniel I.R.

AU - Foshage, Audra M.

AU - Joshi, Piyush

AU - Nitin, Rachana

AU - Uhouse, Michael A.

AU - Di Pardo, Alba

AU - Zhang, Ziyan

AU - Maglione, Vittorio

AU - Aschner, Michael

AU - Bowman, Aaron B.

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N2 - The molecular etiology linking the pathogenic mutations in the Huntingtin (Htt) gene with Huntington's disease (HD) is unknown. Prior work suggests a role for Htt in neuronal autophagic function and mutant HTT protein disrupts autophagic cargo loading. Reductions in the bioavailability of the essential metal manganese (Mn) are seen in models of HD. Excess cellular Mn impacts autophagic function, but the target and molecular basis of these changes are unknown. Thus, we sought to determine if changes in cellular Mn status impact autophagic processes in a wild-type or mutant Htt-dependent manner. We report that the HD genotype is associated with reduced Mn-induced autophagy and that acute Mn exposure increases autophagosome induction/formation. To determine if a deficit in bioavailable Mn is mechanistically linked to the autophagy-related HD cellular phenotypes, we examined autophagosomes by electron microscopy. We observed that a 24 h 100 uM Mn restoration treatment protocol attenuated an established HD 'cargo-recognition failure' in the STHdh HD model cells by increasing the percentage of filled autophagosomes. Mn restoration had no effect on HTT aggregate number, but a 72 h co-treatment with chloroquine (CQ) in GFP-72Q-expressing HEK293 cells increased the number of visible aggregates in a dose-dependent manner. As CQ prevents autophagic degradation this indicates that Mn restoration in HD cell models facilitates incorporation of aggregates into autophagosomes. Together, these findings suggest that defective Mn homeostasis in HD models is upstream of the impaired autophagic flux and provide proof-of-principle support for increasing bioavailable Mn in HD to restore autophagic function and promote aggregate clearance.

AB - The molecular etiology linking the pathogenic mutations in the Huntingtin (Htt) gene with Huntington's disease (HD) is unknown. Prior work suggests a role for Htt in neuronal autophagic function and mutant HTT protein disrupts autophagic cargo loading. Reductions in the bioavailability of the essential metal manganese (Mn) are seen in models of HD. Excess cellular Mn impacts autophagic function, but the target and molecular basis of these changes are unknown. Thus, we sought to determine if changes in cellular Mn status impact autophagic processes in a wild-type or mutant Htt-dependent manner. We report that the HD genotype is associated with reduced Mn-induced autophagy and that acute Mn exposure increases autophagosome induction/formation. To determine if a deficit in bioavailable Mn is mechanistically linked to the autophagy-related HD cellular phenotypes, we examined autophagosomes by electron microscopy. We observed that a 24 h 100 uM Mn restoration treatment protocol attenuated an established HD 'cargo-recognition failure' in the STHdh HD model cells by increasing the percentage of filled autophagosomes. Mn restoration had no effect on HTT aggregate number, but a 72 h co-treatment with chloroquine (CQ) in GFP-72Q-expressing HEK293 cells increased the number of visible aggregates in a dose-dependent manner. As CQ prevents autophagic degradation this indicates that Mn restoration in HD cell models facilitates incorporation of aggregates into autophagosomes. Together, these findings suggest that defective Mn homeostasis in HD models is upstream of the impaired autophagic flux and provide proof-of-principle support for increasing bioavailable Mn in HD to restore autophagic function and promote aggregate clearance.

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