Skip to main content
SpringerLink
Log in

Gedunin Degrades Aggregates of Mutant Huntingtin Protein and Intranuclear Inclusions via the Proteasomal Pathway in Neurons and Fibroblasts from Patients with Huntington’s Disease

  • Original Article
  • Published:
Neuroscience Bulletin Aims and scope Submit manuscript

Abstract

Huntington’s disease (HD) is a deadly neurodegenerative disease with abnormal expansion of CAG repeats in the huntingtin gene. Mutant Huntingtin protein (mHTT) forms abnormal aggregates and intranuclear inclusions in specific neurons, resulting in cell death. Here, we tested the ability of a natural heat-shock protein 90 inhibitor, Gedunin, to degrade transfected mHTT in Neuro-2a cells and endogenous mHTT aggregates and intranuclear inclusions in both fibroblasts from HD patients and neurons derived from induced pluripotent stem cells from patients. Our data showed that Gedunin treatment degraded transfected mHTT in Neuro-2a cells, endogenous mHTT aggregates and intranuclear inclusions in fibroblasts from HD patients, and in neurons derived from induced pluripotent stem cells from patients in a dose- and time-dependent manner, and its activity depended on the proteasomal pathway rather than the autophagy route. These findings also showed that although Gedunin degraded abnormal mHTT aggregates and intranuclear inclusions in cells from HD patient, it did not affect normal cells, thus providing a new perspective for using Gedunin to treat HD.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. MacDonald ME, Ambrose CM, Duyao MP, Myers RH, Lin C, Srinidhi L, et al. A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington’s disease chromosomes. Cell 1993, 72: 971–983.

    Article  Google Scholar 

  2. Kieburtz K, Reilmann R, Olanow CW. Huntington’s disease: current and future therapeutic prospects. Mov Disord 2018, 33: 1033–1041.

    Article  Google Scholar 

  3. Squitieri F, Griguoli A, Capelli G, Porcellini A, D’Alessio B. Epidemiology of Huntington disease: first post-HTT gene analysis of prevalence in Italy. Clin Genet 2016, 89: 367–370.

    Article  CAS  Google Scholar 

  4. Louis ED, Lee P, Quinn L, Marder K. Dystonia in Huntington’s disease: prevalence and clinical characteristics. Mov Disord 1999, 14: 95–101.

    Article  CAS  Google Scholar 

  5. Walker FO. Huntington’s disease. Semin Neurol 2007, 27: 143–150.

    Article  Google Scholar 

  6. Slaughter JR, Martens MP, Slaughter KA. Depression and Huntington’s disease: prevalence, clinical manifestations, etiology, and treatment. CNS Spectr 2001, 6: 306–326.

    Article  CAS  Google Scholar 

  7. Orth M, Handley OJ, Schwenke C, Dunnett SB, Craufurd D, Ho AK, et al. Observing Huntington’s disease: the European Huntington’s disease network’s REGISTRY. PLoS Curr 2010, 2: RRN1184.

    PubMed  Google Scholar 

  8. Aylward EH, Nopoulos PC, Ross CA, Langbehn DR, Pierson RK, Mills JA, et al. Longitudinal change in regional brain volumes in prodromal Huntington disease. J Neurol Neurosurg Psychiatry 2011, 82: 405–410.

    Article  Google Scholar 

  9. Saudou F, Humbert S. The biology of Huntingtin. Neuron 2016, 89: 910–926.

    Article  CAS  Google Scholar 

  10. Li HL, Zhang YB, Wu ZY. Development of research on Huntington disease in China. Neurosci Bull 2017, 33: 312–316.

    Article  Google Scholar 

  11. DiFiglia M, Sapp E, Chase KO, Davies SW, Bates GP, Vonsattel JP, et al. Aggregation of Huntingtin in neuronal intranuclear inclusions and dystrophic neurites in brain. Science 1997, 277: 1990–1993.

    Article  CAS  Google Scholar 

  12. Lee JM, Ramos EM, Lee JH, Gillis T, Mysore JS, Hayden MR, et al. CAG repeat expansion in Huntington disease determines age at onset in a fully dominant fashion. Neurology 2012, 78: 690–695.

    Article  CAS  Google Scholar 

  13. Zhang WY, Gu ZL, Liang ZQ, Qin ZH. Mitochondrial dysfunction and Huntington disease. Neurosci Bull 2006, 22: 129–136.

    CAS  PubMed  Google Scholar 

  14. Fisher ER, Hayden MR. Multisource ascertainment of Huntington disease in Canada: prevalence and population at risk. Mov Disord 2014, 29: 105–114.

    Article  Google Scholar 

  15. Taipale M, Jarosz DF, Lindquist S. HSP90 at the hub of protein homeostasis: emerging mechanistic insights. Nat Rev Mol Cell Biol 2010, 11: 515–528.

    Article  CAS  Google Scholar 

  16. Whitesell L, Lindquist SL. HSP90 and the chaperoning of cancer. Nat Rev Cancer 2005, 5: 761–772.

    Article  CAS  Google Scholar 

  17. Baldo B, Weiss A, Parker CN, Bibel M, Paganetti P, Kaupmann K. A Screen for enhancers of clearance identifies Huntingtin as a heat shock protein 90 (Hsp90) client protein. J Biol Chem 2012, 287: 1406–1414.

    Article  CAS  Google Scholar 

  18. Luo WJ, Sun WL, Taldone T, Rodina A, Chiosis G. Heat shock protein 90 in neurodegenerative diseases. Mol Neurodegener 2010, 5: 24.

    Article  Google Scholar 

  19. He WT, Xue W, Gao YG, Hong JY, Yue HW, Jiang LL, et al. HSP90 recognizes the N-terminus of Huntingtin involved in regulation of Huntingtin aggregation by USP19. Sci Rep 2017, 7: 14797.

    Article  Google Scholar 

  20. Tokui K, Adachi H, Waza M, Katsuno M, Minamiyama M, Doi H, et al. 17-DMAG ameliorates polyglutamine-mediated motor neuron degeneration through well-preserved proteasome function in an SBMA model mouse. Hum Mol Genet 2009, 18: 898–910.

    Article  CAS  Google Scholar 

  21. Patwardhan CA, Fauq A, Peterson LB, Miller C, Blagg BS, Chadli A. Gedunin inactivates the co-chaperone p23 protein causing cancer cell death by apoptosis. J Biol Chem 2013, 288: 7313–7325.

    Article  CAS  Google Scholar 

  22. Ma L, Hu B, Liu Y, Vermilyea Scott C, Liu H, Gao L, et al. Human embryonic stem cell-derived gaba neurons correct locomotion deficits in quinolinic acid-lesioned mice. Cell Stem Cell 2012, 10: 455–464.

    Article  CAS  Google Scholar 

  23. van Hagen M, Piebes DGE, de Leeuw WC, Vuist IM, van Roon-Mom WMC, Moerland PD, et al. The dynamics of early-state transcriptional changes and aggregate formation in a Huntington’s disease cell model. BMC Genomics 2017, 18: 373.

    Article  Google Scholar 

  24. Legleiter J, Lotz GP, Miller J, Ko J, Ng C, Williams GL, et al. Monoclonal antibodies recognize distinct conformational epitopes formed by polyglutamine in a mutant Huntingtin fragment. J Biol Chem 2009, 284: 21647–21658.

    Article  CAS  Google Scholar 

  25. Weiss A, Grueninger S, Abramowski D, Giorgio FP, Lopatin MM, Rosas HD, et al. Microtiter plate quantification of mutant and wild-type Huntingtin normalized to cell count. Anal Biochem 2011, 410: 304–306.

    Article  CAS  Google Scholar 

  26. Rubinsztein DC. The roles of intracellular protein-degradation pathways in neurodegeneration. Nature 2006, 443: 780–786.

    Article  CAS  Google Scholar 

  27. Button RW, Luo SQ, Rubinsztein DC. Autophagic activity in neuronal cell death. Neurosci Bull 2015, 31: 382–394.

    Article  Google Scholar 

  28. Lee DH, Goldberg AL. Proteasome inhibitors: valuable new tools for cell biologists. Trends Cell Biol 1998, 8: 397–403.

    Article  CAS  Google Scholar 

  29. Shacka JJ, Klocke BJ, Roth KA. Autophagy, bafilomycin and cell death: the “a-B-cs” of plecomacrolide-induced neuroprotection. Autophagy 2006, 2: 228–230.

    Article  CAS  Google Scholar 

  30. Brooks E, Arrasate M, Cheung K, Finkbeiner SM. Using antibodies to analyze polyglutamine stretches. Methods Mol Biol 2004, 277: 103–128.

    CAS  PubMed  Google Scholar 

  31. Gutekunst CA, Li SH, Yi H, Mulroy JS, Kuemmerle S, Jones R, et al. Nuclear and neuropil aggregates in Huntington’s disease: relationship to neuropathology. J Neurosci 1999, 19: 2522–2534.

    Article  CAS  Google Scholar 

  32. Gasset-Rosa F, Chillon-Marinas C, Goginashvili A, Atwal RS, Artates JW, Tabet R, et al. Polyglutamine-expanded Huntingtin exacerbates age-related disruption of nuclear integrity and nucleocytoplasmic transport. Neuron 2017, 94: 48–57e44.

    Google Scholar 

  33. Rodriguez-Lebron E, Paulson HL. Allele-specific RNA interference for neurological disease. Gene Ther 2006, 13: 576–581.

    Article  CAS  Google Scholar 

  34. Aguiar S, van der Gaag B, Cortese FAB. RNAi mechanisms in Huntington’s disease therapy: siRNA versus shRNA. Transl Neurodegener 2017, 6: 30.

    Article  Google Scholar 

  35. Ihry RJ, Worringer KA, Salick MR, Frias E, Ho D, Theriault K, et al. p53 inhibits CRISPR-Cas9 engineering in human pluripotent stem cells. Nat Med 2018, 24: 939–946.

    Article  CAS  Google Scholar 

  36. Yang S, Chang R, Yang H, Zhao T, Hong Y, Kong HE, et al. CRISPR/Cas9-mediated gene editing ameliorates neurotoxicity in mouse model of Huntington’s disease. J Clin Invest 2017, 127: 2719–2724.

    Article  Google Scholar 

  37. Haapaniemi E, Botla S, Persson J, Schmierer B, Taipale J. CRISPR-Cas9 genome editing induces a p53-mediated DNA damage response. Nat Med 2018, 24: 927.

    Article  CAS  Google Scholar 

  38. Becher MW, Kotzuk JA, Sharp AH, Davies SW, Bates GP, Price DL, et al. Intranuclear neuronal inclusions in Huntington’s disease and dentatorubral and pallidoluysian atrophy: correlation between the density of inclusions and IT15 CAG triplet repeat length. Neurobiol Dis 1998, 4: 387–397.

    Article  CAS  Google Scholar 

  39. Duyao MP, Auerbach AB, Ryan A, Persichetti F, Barnes GT, McNeil SM, et al. Inactivation of the mouse Huntington’s disease gene homolog Hdh. Science 1995, 269: 407–410.

    Article  CAS  Google Scholar 

  40. Flierl A, Oliveira LM, Falomir-Lockhart LJ, Mak SK, Hesley J, Soldner F, et al. Higher vulnerability and stress sensitivity of neuronal precursor cells carrying an alpha-synuclein gene triplication. PLoS One 2014, 9: e112413.

    Article  Google Scholar 

Download references

Acknowledgements

We thank all members of the HD families for trusting us and supporting us to the end of this work. We also thank Dr. Su-Chun Zhang for helping with this project. This work was supported by the National Key Research and Development Program of China (2018YFA0108004) and the National Natural Science Foundation of China (81271259).

Author information

Author notes

  1. Weiqi Yang, Jingmo Xie, Qiang Qiang and Li Li have contributed equally to this work.

Authors and Affiliations

  1. Department of Anatomy, Histology and Embryology, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, China

    Weiqi Yang, Qiong Liu, Guomin Zhou & Lixiang Ma

  2. School of Medical College, Hexi University, Zhangye, 734000, China

    Weiqi Yang

  3. Department of Anatomy, Histology and Embryology, Zhaoqing Medical College, Zhaoqing, 526000, China

    Jingmo Xie

  4. Department of Neurology, Huadong Hospital, Fudan University, Shanghai, 200040, China

    Qiang Qiang, Yiqing Ren, Wenlei Ren & Wenshi Wei

  5. Department of Anesthesiology, Huashan Hospital, Fudan University, Shanghai, 200040, China

    Li Li

  6. Department of Neurology and Institute of Neurology, First Affiliated Hospital, Fujian Medical University, Fuzhou, 350005, China

    Xiang Lin

  7. School of Life Sciences, Fudan University, Shanghai, 200438, China

    Hexige Saiyin

Authors
  1. Weiqi Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jingmo Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qiang Qiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Li Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xiang Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yiqing Ren
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Wenlei Ren
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Qiong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Guomin Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Wenshi Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Hexige Saiyin
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Lixiang Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Hexige Saiyin or Lixiang Ma.

Ethics declarations

Conflict of interest

All authors claim that there are no conflicts of interest.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 486 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yang, W., Xie, J., Qiang, Q. et al. Gedunin Degrades Aggregates of Mutant Huntingtin Protein and Intranuclear Inclusions via the Proteasomal Pathway in Neurons and Fibroblasts from Patients with Huntington’s Disease. Neurosci. Bull. 35, 1024–1034 (2019). https://doi.org/10.1007/s12264-019-00421-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12264-019-00421-5

Keywords

Search

Navigation