Abstract
Huntington’s disease (HD) is an autosomal dominant neurodegenerative disorder caused by toxic aggregates of mutant huntingtin protein (mHTT) in the brain. Decreasing mHTT is a potential strategy for therapeutic purpose of HD. Valosin-containing protein (VCP/p97) is a crucial regulator of proteostasis, which regulates the degradation of damaged protein through proteasome and autophagy pathway. Since VCP has been implicated in pathogenesis of HD as well as other neurodegenerative diseases, small molecules that specifically regulate the activity of VCP may be of therapeutic benefits for HD patients. In this study we established a high-throughput screening biochemical assay for VCP ATPase activity measurement and identified gossypol, a clinical approved drug in China, as a novel modulator of VCP. Gossypol acetate dose-dependently inhibited the enzymatic activity of VCP in vitro with IC50 of 6.53±0.6 μM. We further demonstrated that gossypol directly bound to the interface between the N and D1 domains of VCP. Gossypol acetate treatment not only lowered mHTT levels and rescued HD-relevant phenotypes in HD patient iPS-derived Q47 striatal neurons and HD knock-in mouse striatal cells, but also improved motor function deficits in both Drosophila and mouse HD models. Taken together, gossypol acetate acted through a gain-of-function way to induce the formation of VCP-LC3-mHTT ternary complex, triggering autophagic degradation of mHTT. This study reveals a new strategy for treatment of HD and raises the possibility that an existing drug can be repurposed as a new treatment of neurodegenerative diseases.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (31270830 and 21572038 to YJD; 81870990 to BXL;81625022, 91853205, 81821005 to CL; 31970748 to YHF), the Science and Technology Commission of Shanghai Municipality (18431907100 and 19XD1404700 to C.L.), the Development Fund for Shanghai Talents, Fund of State Key Laboratory of Bioorganic and Natural Products Chemistry, and Fund of State Key Laboratory of Drug Research and Chinese Academy of Science (SIMM1601KF-08). The authors acknowledge instructive advice and support from Prof. Fang Huang from Fudan University and her student Yu-fang Yang, Prof. Jun O. Liu from the Johns Hopkins University and Prof. Jian-zhong Yu from the University of Kansas.
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XJL performed the ATPase assay, PTS assay, pull-down assay, Western blot analysis, mouse behavioral experiments and the corresponding data analysis, and wrote the manuscript. YYZ contributed to manuscript writing and modification and was involved in the mouse behavioral experiments. YHF contributed to the HTRF assay, neuronal loss, and apoptosis measurement. HZ contributed to simulating the interaction model of GA and VCP. HXL completed Drosophila model behavior experiments. QFL was involved in performing the Western blot analysis. HLL was involved in the mouse behavioral experiments. RKT conducted the HTS. CXJ performed partial proteolysis assays and ITC experiments. WJ and ZXL were involved in protein expression and purification, ITC, and mouse behavioral experiments. CL conceived and supervised the simulation of the interaction model. BXL conceived and supervised the HD cell and animal experiments. YJD conceived and supervised the project and modified the manuscripts.
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Li, Xj., Zhang, Yy., Fu, Yh. et al. Gossypol, a novel modulator of VCP, induces autophagic degradation of mutant huntingtin by promoting the formation of VCP/p97-LC3-mHTT complex. Acta Pharmacol Sin 42, 1556–1566 (2021). https://doi.org/10.1038/s41401-020-00605-0
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DOI: https://doi.org/10.1038/s41401-020-00605-0
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